CAL-T CONSTRUCTS AND USES THEREOF

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation under 35 U.S.C. of § 120 of co-pending U.S. application Ser. No. 17/072,490 filed Oct. 16, 2020 and which claims benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 62/916,924 filed Oct. 18, 2019, the contents of which are incorporated herein by reference in their entireties.

SEQUENCE LISTING

The instant application contains a Sequence Listing that has been submitted in XML format via Patent Center and is hereby incorporated by reference in its entirety. Said XML copy, created on Jun. 15, 2023, is named “030258-096030USC1_SL.xml” and is 2,487,922 bytes in size.

TECHNICAL FIELD

The technology described herein relates to chimeric antigen ligand (CAL) technology, e.g., for treating of autoimmune and/or T-cell mediated conditions.

BACKGROUND

The major cause of autoimmunity and transplant rejection, e.g., following an organ transplant, is autoreactive and alloreactive T cells attacking the patient's or donor's organ(s), tissue, and/or cells. Current treatments for preventing autoimmunity and transplant rejection involve stringent immunosuppression, which can lead to severe, unwanted side effects including extreme susceptibility to infection, malignant and benign neoplasms, multiple organ and systems failure. In order to prevent autoimmunity and transplant rejection without inducing unwanted side effects, it would be advantageous to delete autoreactive and alloreactive T cells in a patient receiving an organ transplant without needing to additionally administer immunosuppressant medications, or to reduce the doses of the immunosuppressant regime delivered concurrently. This type of therapy, however, has not been successfully demonstrated.

T cells recognize their target cells by using receptors on their cell surface which are called T Cell Receptors (TCRs). TCRs have a recognition portion and a signaling portion. When the recognition portion binds to the natural complexes formed in the body in the presence of a diseased cell, the signaling portion is activated, which leads to the T cell engaging in killing activity or recruiting other immune cells to destroy the diseased cell. The natural complexes are also known as peptide-major histocompatibility complexes (pMHC complexes). CAR-T cell therapy is known and seeks to help T cells recognize autoreactive and alloreactive T cells. This is accomplished by genetically altering a T cell so that it expresses a chimeric antigen receptor (CAR). The CAR is an altered TCR, in which the natural recognition portion is removed and replaced with a synthetic recognition portion that is designed to more effectively recognize the autoreactive and alloreactive T cells by very specifically detecting the presence of a molecule unique to the autoreactive and alloreactive T cells. These CAR-T cells are then given to a patient. Inside the patient, their synthetic CAR molecules will bind to the autoreactive and alloreactive T cells and in the act of that binding, activate the T cell, resulting in the patient's own immune system attacking the diseased T-cells.

SUMMARY

Described herein are methods and compositions relating to chimeric antigen ligands (CAL). The CALs described herein comprise a TCR recognition domain and a biomolecular interaction domain. The TCR recognition permits the CAL to bind to autoreactive and/or alloreactive T cells in an antigen specific manner. The biomolecular interaction domain permits an immune killer cell (e.g., a NK cell, a T cell, or a dendritic cell) to bind to the CAL, thereby promoting killing of the autoreactive and/or alloreactive T cell by the immune killer cell.

The TCR recognition domain of a CAL binds specifically to a TCR, e.g., a TCR expressed on the surface of an autoreactive and/or alloreactive T cell. Exemplary but non-limiting TCR recognition domains include peptide-MHC complexes, e.g., in monomeric, oligomeric, or multimeric form. The TCR recognition domain can comprise natural or synthetic sequences. Specific examples of peptide-MHC complexes that include autoreactive or alloreactive antigens are provided elsewhere herein.

The biomolecular interaction domain of the CAL permits specific binding of the CAL with a second biomolecule, e.g., a receptor on the immune killer cell. In some embodiments, the biomolecular interaction domain of the CAL is recognized by an endogenous receptor on the immune killer cell. In some embodiments, the biomolecular interaction domain of the CAL is recognized by an engineered receptor on the immune killer cell. Exemplary but non-limiting biomolecular interaction domains include FITC (which can be recognized by a FITC CAR-T cell system), a leucine zipper domain, a zinc finger domain, PSD95-Dlg1-zo-1 (PDZ) domains, a streptavidin domain and a streptavidin binding protein (SBP) domain, a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1), a PYL domain and an ABI domain, a chemically-induced pair of interaction domains as described elsewhere herein, a Snap-tag, a Halo tag, a T14-3-3-cdeltaC and/or a C-Terminal peptide of PMA2 (CT52), a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein a Fab domain, and/or an anti-CD3 domain.

As noted, the CALs described herein can be used in combination with CAR-T systems. Exemplary but non-limiting CAR-T systems suitable for use with the methods and compositions described herein include SUPRA CAR and SPLIT CAR. CAR-T systems are discussed in more detail elsewhere herein and are known in the art.

In one aspect of any of the embodiments, described herein is a composition, comprising: a) a TCR recognition domain; and one or both of: b) an intracellular signaling domain; and c) a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain).

In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; wherein the first-type and second-type biomolecular interaction domains bind specifically to each other. In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the first-type and third-type biomolecular interaction domains bind specifically to each other. In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; and a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the second-type and third-type biomolecular interaction domains compete for binding to the first-type biomolecular interaction domain. In some embodiments of any of the aspects, the third-type biomolecular interaction domain and first-type biomolecular interaction domain have a higher affinity for each other than the second-type biomolecular interaction domain and first-type biomolecular interaction domain.

In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; a signaling polypeptide comprising a second-type biomolecular interaction domain, a fourth-type biomolecular interaction domain, and an intracellular signaling domain; and a recognition polypeptide comprising a second recognition domain and a fifth-type biomolecular interaction domain; wherein the first-type biomolecular interaction domain and the second-type biomolecular interaction domain bind specifically to each other; and wherein the fourth-type biomolecular interaction domain and the fifth-type biomolecular interaction domain bind specifically to each other. In some embodiments of any of the aspects, the fourth-type biomolecular interaction domain and fifth-type biomolecular interaction domain have a weaker affinity than the second-type biomolecular interaction domain and first-type protein interaction domain. In some embodiments of any of the aspects, the first polypeptide further comprises a sixth-type biomolecular interaction domain and the recognition polypeptide further comprises a seventh-type biomolecular interaction domain which bind specifically to each other.

In some embodiments of any of the aspects, the first polypeptide comprises the entire TCR recognition domain. In some embodiments of any of the aspects, the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain. In some embodiments of any of the aspects, the TCR recognition domain comprises a non-polypeptide component.

In some embodiments of any of the aspects, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain. In some embodiments of any of the aspects, the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.

In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; featureless peptide MHC; or a MHC-peptide fusion. In some embodiments of any of the aspects, the peptide is a human or non-human peptide. In some embodiments of any of the aspects, the peptide is a Minor Histocompatibility Antigen (MiHA). In some embodiments of any of the aspects, the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide complex is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC is a MHC class I or a MHC class II.

In some embodiments of any of the aspects, the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion. In some embodiments of any of the aspects, the CD1 is CD1d.

In some embodiments of any of the aspects, the biomolecular interaction domains are found on an extracellular portion of the respective polypeptides. In some embodiments of any of the aspects,

• • a. wherein the biomolecular interaction domain(s) is a leucine zipper, or any binding pair of biomolecular interaction domains are collectively a pair of leucine zippers;
  • b. wherein the biomolecular interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE);
  • c. wherein the biomolecular interaction domain(s) is a PSD95-Dlg1-zo-1 (PDZ) domain;
  • d. wherein the biomolecular interaction domain(s) is a streptavidin and/or a streptavidin binding biomolecular (SBP) or any binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding biomolecular (SBP);
  • e. wherein the biomolecular interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding biomolecular (FKBP);
  • f. wherein the biomolecular interaction domain(s) is a cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506 binding biomolecular (FKBP);
  • g. wherein the biomolecular interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding biomolecular (FKBP);
  • h. wherein the biomolecular interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1);
  • i. wherein the biomolecular interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag;
  • j. wherein the biomolecular interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52);
  • k. wherein the biomolecular interaction domain(s) is a PYL and/or a ABI, or any binding pair of biomolecular interaction domains are collectively a PYL and a ABI;
  • l. wherein the biomolecular interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain;
  • m. wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags;
  • n. wherein the biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding biomolecular or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or
  • o. wherein the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE binding protein.
    In some embodiments of any of the aspects, the nucleotide tag is a DNA tag or dsDNA tag.

In some embodiments of any of the aspects, the intracellular signaling domain comprises or is a signaling domain from one or more proteins selected from the group consisting of: TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.

In one aspect of any of the embodiments, described herein is a cell comprising and/or expressing a composition described herein. In one aspect of any of the embodiments, described herein is a composition comprising a first polypeptide of any of the preceding claims and a cell expressing or comprising the signaling polypeptide of any of the preceding claims.

In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any of the aspects, the TCR recognition domain comprises a synthetic MHC. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.

In some embodiments of any of the aspects, the cell is a NK cell, dendritic cell, regulatory T cell, or effector T cell. In some embodiments of any of the aspects, the cell is engineered to express one of more of the polypeptide(s) of the composition. In some embodiments of any of the aspects, the cell is engineered to express the signaling polypeptide of the composition. In some embodiments of any of the aspects, the cell is further engineered to knockout or knockdown the native MHCI/II. In some embodiments of any of the aspects, the cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.

In one aspect of any of the embodiments, described herein is a composition, comprising a TCR recognition domain and an intracellular signaling domain but not comprising a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain). In one aspect of any of the embodiments, described herein is a composition, comprising a TCR recognition domain and a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain) and but not comprising an intracellular signaling domain.

In one aspect of any of the embodiments, described herein is a method of treating or preventing an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD in a subject in need thereof, the method comprising administering to the subject a composition and/or cell of any of the preceding claims. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC allogeneic to the subject.

In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC autologous to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC xenogeneic to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic to the subject. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells. In some embodiments of any of the aspects, the transplant is any human or non-human cell, tissue, or organ. In some embodiments of any of the aspects, the transplant is an allogeneic hematopoietic stem cell or solid organ transplantation.

In some embodiments of any of the aspects, the malignant T cell condition is T cell acute lymphoblastic leukemia or T cell lymphoblastic lymphoma.

In some embodiments of any of the aspects, the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma. In some embodiments of any of the aspects, the autoimmune disease is thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis), palindromic arthritis, inflammatory hyperproliferative skin diseases, psoriasis such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and psoriasis of the nails, atopy including atopic diseases such as hay fever and Job's syndrome, dermatitis including contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, non-specific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic intraocular inflammatory diseases, urticaria such as chronic allergic urticaria and chronic idiopathic urticaria, including chronic autoimmune urticaria, myositis, polymyositis/dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic scleroderma), sclerosis such as systemic sclerosis, multiple sclerosis (MS) such as spino-optical MS, primary progressive MS (PPMS), and relapsing remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, sclerosis disseminata, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (for example, Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis such as ulcerative colitis, colitis ulcerosa, microscopic colitis, collagenous colitis, colitis polyposa, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), bowel inflammation, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome, including adult or acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, an autoimmune hematological disorder, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as in meningitis, herpes gestationis, pemphigoid gestationis, pruritis scroti, autoimmune premature ovarian failure, sudden hearing loss due to an autoimmune condition, IgE-mediated diseases such as anaphylaxis and allergic and atopic rhinitis, encephalitis such as Rasmussen's encephalitis and limbic and/or brainstem encephalitis, uveitis, such as anterior uveitis, acute anterior uveitis, granulomatous uveitis, nongranulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome such as chronic or acute glomerulonephritis such as primary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membrano- or membranous proliferative GN (MPGN), including Type I and Type II, and rapidly progressive GN, proliferative nephritis, autoimmune polyglandular endocrine failure, balanitis including balanitis circumscripta plasmacellularis, balanoposthitis, erythema annulare centrifugum, erythema dyschromicum perstans, eythema multiform, granuloma annulare, lichen nitidus, lichen sclerosus et atrophicus, lichen simplex chronicus, lichen spinulosus, lichen planus, lamellar ichthyosis, epidermolytic hyperkeratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, allergic reaction, eczema including allergic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and vesicular palmoplantar eczema, asthma such as asthma bronchiale, bronchial asthma, and auto-immune asthma, conditions involving infiltration of T cells and chronic inflammatory responses, immune reactions against foreign antigens such as fetal A-B-O blood groups during pregnancy, chronic pulmonary inflammatory disease, autoimmune myocarditis, leukocyte adhesion deficiency, lupus, including lupus nephritis, lupus cerebritis, pediatric lupus, non-renal lupus, extra-renal lupus, discoid lupus and discoid lupus erythematosus, alopecia lupus, systemic lupus erythematosus (SLE) such as cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and lupus erythematosus disseminatus, juvenile onset (Type I) diabetes mellitus, including pediatric insulin-dependent diabetes mellitus (IDDM), adult onset diabetes mellitus (Type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic large-artery disorder, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, sarcoidosis, granulomatosis including lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitides, including vasculitis, large-vessel vasculitis (including polymyalgia rheumatica and giant-cell (Takayasu's) arteritis), medium-vessel vasculitis (including Kawasaki's disease and polyarteritis nodosa/periarteritis nodosa), microscopic polyarteritis, immunovasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, necrotizing vasculitis such as systemic necrotizing vasculitis, and ANCA-associated vasculitis, such as Churg-Strauss vasculitis or syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs positive anemia, Diamond Blackfan anemia, hemolytic anemia or immune hemolytic anemia including autoimmune hemolytic anemia (AIHA), pernicious anemia (anemia perniciosa), Addison's disease, pure red cell anemia or aplasia (PRCA), Factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases involving leukocyte diapedesis, CNS inflammatory disorders, multiple organ injury syndrome such as those secondary to septicemia, trauma or hemorrhage, antigen-antibody complex-mediated diseases, anti-glomerular basement membrane disease, anti-phospholipid antibody syndrome, allergic neuritis, Behcet's disease/syndrome, Castleman's syndrome, Goodpasture's syndrome, Reynaud's syndrome, Sjogren's syndrome, Stevens-Johnson syndrome, pemphigoid such as pemphigoid bullous and skin pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, pemphigus mucus-membrane pemphigoid, and pemphigus erythematosus), autoimmune polyendocrinopathies, Reiter's disease or syndrome, an immune complex disorder such as immune complex nephritis, antibody-mediated nephritis, polyneuropathies, chronic neuropathy such as IgM polyneuropathies or IgM-mediated neuropathy, and autoimmune or immune-mediated thrombocytopenia such as idiopathic thrombocytopenic purpura (ITP) including chronic or acute ITP, scleritis such as idiopathic cerato-scleritis, episcleritis, autoimmune disease of the testis and ovary including autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis such as autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis), or subacute thyroiditis, idiopathic hypothyroidism, Grave's disease, polyglandular syndromes such as autoimmune polyglandular syndromes (or polyglandular endocrinopathy syndromes), paraneoplastic syndromes, including neurologic paraneoplastic syndromes such as Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis such as allergic encephalomyelitis or encephalomyelitis allergica and experimental allergic encephalomyelitis (EAE), myasthenia gravis such as thymoma-associated myasthenia gravis, cerebellar degeneration, neuromyotonia, opsoclonus or opsoclonus myoclonus syndrome (OMS), and sensory neuropathy, multifocal motor neuropathy, Sheehan's syndrome, autoimmune hepatitis, lupoid hepatitis, giant-cell hepatitis, autoimmune chronic active hepatitis, lymphoid interstitial pneumonitis (LIP), bronchiolitis obliterans (non-transplant) vs NSIP, Guillain-Barre syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis such as primary biliary cirrhosis and pneumonocirrhosis, autoimmune enteropathy syndrome, Celiac or Coeliac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, amylotrophic lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear disease such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsed or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome/nonsyphilitic interstitial keratitis, Bell's palsy, Sweet's disease/syndrome, rosacea autoimmune, zoster-associated pain, amyloidosis, a non-cancerous lymphocytosis, a primary lymphocytosis, which includes monoclonal B cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathy, paraneoplastic syndrome, channelopathies including channelopathies of the CNS, autism, inflammatory myopathy, focal or segmental or focal segmental glomerulosclerosis (FSGS), endocrine opthalmopathy, uveoretinitis, chorioretinitis, autoimmune hepatological disorder, fibromyalgia, multiple endocrine failure, Schmidt's syndrome, adrenalitis, gastric atrophy, presenile dementia, demyelinating diseases such as autoimmune demyelinating diseases and chronic inflammatory demyelinating polyneuropathy, Dressler's syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility, e.g., due to anti-spermatozoan antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortion, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, bird-fancier's lung, allergic granulomatous angiitis, benign lymphocytic angiitis, Alport's syndrome, alveolitis such as allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reaction, Sampter's syndrome, Caplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial lung fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum et diutinum, erythroblastosis fetalis, eosinophilic faciitis, Shulman's syndrome, Felty's syndrome, cyclitis such as chronic cyclitis, heterochronic cyclitis, iridocyclitis (acute or chronic), or Fuch's cyclitis, Henoch-Schonlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndromes, Evan's syndrome, autoimmune gonadal failure, Sydenham's chorea, post-streptococcal nephritis, thromboangitis ubiterans, thyrotoxicosis, tabes dorsalis, chorioiditis, giant-cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplant organ reperfusion, retinal autoimmunity, aphthae, aphthous stomatitis, arteriosclerotic disorders, aspermiogenesis, autoimmune hemolysis, Boeck's disease, enteritis allergica, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, febris rheumatica, Hamman-Rich's disease, sensoneural hearing loss, ileitis regionalis, leucopenia, transverse myelitis, primary idiopathic myxedema, ophthalmia symphatica, polyradiculitis acuta, pyoderma gangrenosum, acquired spenic atrophy, vitiligo, toxic-shock syndrome, conditions involving infiltration of T cells, leukocyte-adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, diseases involving leukocyte diapedesis, multiple organ injury syndrome, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathies, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine failure, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid, frontal, maxillary, or sphenoid sinusitis, an eosinophil-related disorder such as eosinophilia, pulmonary infiltration eosinophilia, eosinophilia-myalgia syndrome, Loffler's syndrome, chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, granulomas containing eosinophils, seronegative spondyloarthritides, polyendocrine autoimmune disease, sclerosing cholangitis, sclera, episclera, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia telangiectasia syndrome, angiectasis, autoimmune disorders associated with collagen disease, rheumatism, allergic hypersensitivity disorders, glomerulonephritides, reperfusion injury, ischemic re-perfusion disorder, lymphomatous tracheobronchitis, inflammatory dermatoses, dermatoses with acute inflammatory components, and autoimmune uveoretinitis (AUR).

In some embodiments of any of the aspects, the T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy. In some embodiments of any of the aspects, the biologic, cell-therapy, or gene therapy is an adeno-associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.

In some embodiments of any of the aspects, the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129. In some embodiments of any of the aspects, the disease is vitiligo and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO: 2021. In some embodiments of any of the aspects, the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO: 2039. In some embodiments of any of the aspects, the disease is type 1 diabetes and the TCR recognition domain comprises one or more of SEQ ID NOs: 8-17; comprises HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or comprises HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129. In some embodiments of any of the aspects, the disease is vitiligo and the TCR recognition domain comprises SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or comprises HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or comprises HLA-A*0101 and SEQ ID NO: 2021. In some embodiments of any of the aspects, the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or comprises HLA-B*0702 and SEQ ID NO: 2038; or comprises HLA-B*0801 and SEQ ID NO: 2039.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 demonstrates that pMHC tetramer+FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion or manner. I-H2Kb: MHC class I tetramer. I-Ab−: Control tetramer. Figure discloses SEQ ID NOS 2752-2753, 2750 and 2754, respectively, in order of appearance.

FIG. 2 depicts expression of activation marker CD69 on OTi cells at different time points (24, 48, 72 hrs.). Binding of Jurkat+pMHC to OTi cells does not change Jurkat live count (right). Figure discloses SEQ ID NOS 2750, 2750 and 2754, respectively, in order of appearance.

FIG. 3 depicts the expression of CD69 on Jurkat CAR, e.g., CAL T cells in different time points (24, 48, 72 hrs.). Figure discloses SEQ ID NOS 2750 and 2755, respectively, in order of appearance.

FIG. 4 depicts the cytotoxicity of Primary human CD8 CAR, e.g., CAL T cells with different concentrations of tetramers (left). Minimal activation of target cells (CD69 on CAR, e.g., CAL T cells and target cells) after coculturing with OTi cells (FIG. 2 left). I-Ab: Ctrl/H2Kb: MHC-I. Figure discloses SEQ ID NOS 2750, 2754 and 2750, respectively, in order of appearance.

FIG. 5 demonstrates that the cytotoxicity of human CD8 CAR, e.g., CAL T is highly specific and was not seen with ctrl tetramer (left). No cytotoxicity was seen on killer CAR, e.g., CAL T cells or bystander CD4 T Cells after co-culturing with pMHC and splenocytes. Figure discloses SEQ ID NOS 2754, 2750, 2754, 2750, 2754 and 2750, respectively, in order of appearance.

FIG. 6A depicts a schematic of SUPRA CAR design applied to provide Universal CAL. This design separates the cell targeting molecule module from the killer cell. FIG. 6B depicts that SUPRA CARs can be designed with CD3ζ domain uncoupled from the costimulatory domains to provide Universal CAL as described herein.

FIGS. 7A-7D depict key features of the SUPRA CAR systems that can be applied to Universal CAL. FIG. 7A demonstrates that zipCAR activation is tunable through modulation of zipFv concentration, zipper affinity, scFv affinity, and zipCAR expression level in human primary CD4 T cells, as demonstrated by IFN-g production. FIG. 7B demonstrates that SUPRA CAR system, as applied to Universal CAL, can perform combination antigen detection to form AND gate logic in CD4 T cells. FIG. 7C demonstrates that xenograft animal tumor model shows tumor eradication (as demonstrated by luciferase photon flux given by the tumor cells) by the SUPRA CAR T cells. FIG. 7D demonstrates that SUPRA CARs as applied to Universal CAL can be used to control different cell types, such as CD4 and CD8 T cells, against two different antigens. CD69 expression (a T cell activation marker) is quantified with flow cytometry for CD4 and CD8 T cells. (From Cho, Collins, and Wong, Cell. 2018)

FIG. 8 depicts the timeline of double Hu-PBMC-HSCT-skin graft mouse model generation.

FIG. 9 depicts a summary of double hu-PBMC-HSCT-skin graft mouse model generation.

FIG. 10 depicts key features of pMHC multimer+CAR, e.g., CAL T cell system.

FIG. 11 depicts a table of experimental design. Figure discloses SEQ ID NOS 2750, 2756 and 2754, respectively, in order of appearance.

FIG. 12 demonstrates verification of FITC-conjugated tetramer mediated activation. Figure discloses SEQ ID NOS 2750 and 2751, respectively, in order of appearance.

FIG. 13 depicts a time course of FITC-conjugated tetramer mediated activation. Figure discloses SEQ ID NOS 2750, 2751, 2750, 2751, 2750, 2751, 2750 and 2751, respectively, in order of appearance.

FIG. 14 depicts a graph of Jurkat cell counts. Figure discloses SEQ ID NOS 2750 and 2754, respectively, in order of appearance.

FIG. 15 depicts tetramer staining. Figure discloses SEQ ID NOS 2750, 2754, 2751, 2756, 2754, 2750 and 2754, from left to right and top to bottom.

FIG. 16 depicts heatmaps of indicated staining levels. Figure discloses SEQ ID NOS 2751, 2757, 2754, 2750, 2754, 2751, 2754, 2757, 2750, 2754, 2751, 2754, 2757, 2750 and 2754, respectively, in order of appearance.

FIG. 17 depicts a table of experimental design.

FIGS. 18-20 depict graphs of cytotoxicity levels. FIG. 18 discloses SEQ ID NOS 2754, 2750, 2754 and 2750 from left to right. FIG. 19 discloses SEQ ID NOS 2754 and 2750, respectively, in order of appearance. FIG. 20 discloses SEQ ID NOS 2750, 2754, 2750 and 2754 from left to right.

FIG. 21 depicts the levels of CD69 on OTi CD8 T cells. Figure discloses SEQ ID NOS 2750, 2754, 2750 and 2754 from left to right.

FIG. 22 depicts schematics of two embodiments of the technology described herein.

FIG. 23 depicts a schematic of the FU-CAL embodiments of the technology described herein.

FIG. 24 depicts a schematic of the CAL-BITE embodiments of the technology described herein. The left panel depicts blinatumomab, which is described in more detail in Weiner et al. The Molecular Basis of Cancer 2015 683-694.e3.

FIG. 25 depicts a schematic of the CAL technology disarming autoreactive T cells.

FIGS. 26A-26B depict schematics of T cells design. MHC can be mouse or human. The MiHA can be ovalbumin (against OTi or OTii) or disparate antigens between donor and recipient.

FIG. 27 depicts a graph demonstrating that pMHC tetramer+FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion, while OTii specific tetramer does not.

FIG. 28A depicts expression of CD69 on OTi cells at different time points (24, 48, 72 hrs.).

FIG. 28B is a graph demonstrating that binding of Jurkat+pMHC to OTi cells does not change Jurkat live count.

FIG. 29 depicts a graph demonstrating that cytotoxicity of pMHC-CAR against 1E6 T cell clone exhibits a dose dependent association.

FIG. 30 depicts a graph demonstrating that CAR Jurkat T cells activation (as measured by CD69 surface marker) after exposure to target cells (OTi or OTii TCR expressing T cells) and the corresponding tetramer (OTi or OTii specific) (n=4).

FIG. 31 depicts graphs demonstrating that cytotoxicity of human CD8 CART is highly specific and was not seen with ctrl tetramer. No cytotoxicity was seen on killer CAR T cells or bystander CD4 T Cells after co-culturing with pMHC and splenocytes (n=4).

FIG. 32 depicts a graph demonstrating cytotoxicity of Primary CD8 CART cells against OTi TCR T cells with different concentration of tetramer.

DETAILED DESCRIPTION

Aspects of the invention described herein relate to chimeric antigen ligands. As used herein, “chimeric antigen ligand” or “CAL” refers to an artificially constructed molecule comprising a TCR recognition domain (e.g. an polypeptide comprising at least one MHC sequence as described herein) and at least one biomolecular interaction domain. The TCR recognition domain is selected to bind to specific populations of T cells that it is desirable to target and/or destroy, e.g., for therapeutic purposes in T-cell mediated diseases. In some embodiments, the population of targeted T cells is a population of polyclonal pathogenic T cells. The biomolecular interaction domain is selected to bind to a second cell, e.g., a NK cell, thereby colocalizing the targeted T cell and the second cell and promoting or increasing the inhibition and/or destruction of the targeted T cell. In various embodiments, the CAL is selected to have high affinity or avidity for the TCR, e.g., the TCR variable domain.

The CALs can be used herein with endogenous cells, e.g., in some embodiments, no engineered cells are administered to the subject. In other embodiments, the CALs can be used with engineered cells, e.g., engineered NK cells. In such cases, the engineered cells can comprise one or more CARs, e.g, a CAR comprising an extracellular domain with a biomolecular interaction domain that specifically binds with the biomolecular interaction domain of the CAL.

Accordingly, described herein are chimeric antigen receptors (CARs) in which the recognition and signaling portions of the CAR are separate polypeptides. The two separate polypeptides that make up a complete CAR are able to interact and form the complete CAR by way of protein interaction domains. This permits flexible, modular CAR-T therapy which is capable of complex logic computation, providing a more precise and effective approach to immunotherapy.

In one aspect of any of the embodiments is a CAL and/or chimeric antigen receptor (CAR) having multiple components, and/or a cell or composition comprising a multi-component CAL and/or CAR. Multi-component CALs/CARs are also referred to herein variously as SMART CAL/CAR or SUPRA.

As used herein, traditional “chimeric antigen receptor” or “CAR” refers to an artificially constructed hybrid polypeptide comprising an antigen-binding domain (e.g. an antigen-binding portion of an antibody (e.g. a scFV)) linked to a cell signaling and/or cell activation domain. In some embodiments the cell-signaling domain can be a T-cell signaling domain. In some embodiments, the cell activation domain can be a T-cell activation domain. CARs have the ability to redirect the specificity and reactivity of T cells and other immune cells (e.g., NK cells) toward a selected target in a non-MHC-restricted manner, exploiting the antigen-binding properties of monoclonal antibodies.

The non-MHC-restricted antigen recognition gives T-cells expressing CARs the ability to recognize an antigen independent of antigen processing, thus bypassing a major mechanism of tumor escape. Moreover, when expressed in T-cells, CARs advantageously do not dimerize with endogenous T-cell receptor (TCR) alpha and beta chains. Most commonly, the CAR's extracellular binding domain is composed of a single chain variable fragment (scFv) derived from fusing the variable heavy and light regions of a murine or humanized monoclonal antibody. Alternatively, scFvs may be used that are derived from Fabs (instead of from an antibody, e.g., obtained from Fab libraries), in various embodiments, this scFv is fused to a transmembrane domain and then to an intracellular signaling domain. “First-generation” CARs include those that solely provide CD3zeta signals upon antigen binding, “Second-generation” CARs include those that provide both costimulation (e.g. CD28 or CD 137) and activation (CD3Q). “Third-generation” CARs include those that provide multiple costimulation (e.g. CD28 and CD 137) and activation (CO3Q). In various embodiments, the CAR is selected to have high affinity or avidity for the antigen. Further discussion of CARs can be found, e.g., in Maus et al. Blood 2014 123:2624-35; Reardon et al. Neuro-Oncology 2014 16:1441-1458; Hoyos et al. Haematologica 2012 97:1622; Byrd et al. J Clin Oncol 2014 32:3039-47; Maher et al. Cancer Res 2009 69:4559-4562; and Tamada et al. Clin Cancer Res 2012 18:6436-6445; each of which is incorporated by reference herein in its entirety.

As used herein, “multi-component CAL” refers to a CAL comprising at least two separate polypeptides, neither of which polypeptides is capable of both ligand recognition and signaling activation on its own. As used herein, “multi-component CAR” refers to a CAR comprising at least two separate polypeptides, neither of which polypeptides is capable of both ligand recognition and signaling activation on its own. In some embodiments, the at least two separate polypeptides each comprise a protein interaction domain that permits interaction, e.g., binding of the separate polypeptides. In some embodiments, one of the at least two separate polypeptides is a transmembrane polypeptide having an intracellular T cell receptor (TCR) signaling domain and a second of the at least two separate polypeptides is an extracellular polypeptide having a ligand-binding domain. In some embodiments, a multi-component CAL and/or CAR can comprise two, three, four, five, six, seven, eight, nine, ten or more separate polypeptides.

Various aspects provided herein provide a composition comprising multiple components of a multi-component CAL and/or CAR.

In one aspect of the embodiments is a composition, e.g., a single molecule, comprising a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain. In one aspect of the embodiments is a composition, e.g., a single molecule comprising a TCR recognition domain; and a first-type biomolecular (e.g., protein) interaction domain. Further provided herein is a multi-component CAL and/or CAR comprising a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain. In some embodiments, the composition, e.g, single molecule, comprising a TCR recognition domain; and a first-type biomolecular (e.g., protein) interaction domain does not comprise an antibody, antibody domain, or antibody reagent.

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other. Further provided herein is a multi-component CAL and/or CAR comprising a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other.

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other. Further provided herein is a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain. Further provided herein is a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain.

In various embodiments, the third-type protein interaction domain and first-type protein interaction domain have a higher affinity for each other than the second-type protein interaction domain and first-type protein interaction domain. Affinity can be measured by one skilled in the art using standard methods, for example, by measuring its equilibrium dissociation constant (KO.

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other. Further provided herein is a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

In various embodiments, the fourth-type protein interaction domain and fifth-type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first-type protein interaction domain. Affinity can be measured as described above.

In various embodiments, the first polypeptide further comprises a sixth-type protein interaction domain and the recognition polypeptide further comprises a seventh-type protein interaction domain which bind specifically to each other.

In some embodiments, the first polypeptide comprises the entire TCR recognition domain. In some embodiments, the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.

In some embodiments, a composition described herein can comprise multiple copies or instances of a TCR recognition domain(s), e.g. the TCR recognition domain can be a mulitmer, or oligomer. In some embodiments, a composition described herein can comprise multiple copies or instances of a first polypeptide as described herein.

In various embodiments, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain. In one embodiment, the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.

As used herein, “TCR recognition domain” refers to a domain or portion of a polypeptide that can target or bind specifically to a TCR, e.g., a TCR expressed on the surface of T cell. In some embodiments, the TCR recognition domain can be a TCR variable region (TCR-VR) recognition domain, i.e. is can target or bind specifically to the variable region of a TCR, e.g., a TCR expressed on the surface of a T cell. The TCR recognition domain sequence can be autologous, allogeneic, or xenogeneic to a given subject. In some embodiments, the TCR recognition domain sequence is a wild-type protein or sequence. In some embodiments, the TCR recognition domain sequence is a naturally-occurring variant, e.g., an allele of a wild-type protein or sequence. In some embodiments, the TCR recognition domain sequence is modified relative to a wild-type protein, e.g. chemically modified. In some embodiments, the TCR recognition domain sequence is a derivative and/or variant of a wild-type sequence. In some embodiments, the TCR recognition domain sequence can be a human, or non-human sequence.

A TCR recognition domain can comprise a MHC polypeptide, a MHC polypeptide sequence, and/or comprise a portion of a MHC sequence. The MHC and/or MHC sequence can be autologous, allogeneic, or xenogeneic to a given subject. In some embodiments, the MHC and/or MHC sequence is a wild-type protein or sequence. In some embodiments, the MHC and/or MHC sequence is a naturally-occurring variant, e.g., an allele of MHC. In some embodiments, the MHC and/or MHC sequence is modified relative to a wild-type protein, e.g. chemically modified. In some embodiments, the MHC and/or MHC sequence is a derivative and/or variant of a wild-type MHC sequence. In some embodiments, the MHC and/or MHC sequence can be or comprise a human, or non-human sequence.

In one embodiment, the TCR recognition domain comprises a MHC (Major Histocompatibility Complex), a MHC-peptide complex, or a MHC-peptide fusion. In some embodiments, the TCR recognition domain can comprise a featureless peptide MHC, or a MHC without peptides, or any other molecule that can target or bind specifically to the variable region of the TCR.

The MHC, which is also referred to as the human leukocyte antigen (HLA), is comprised of a set of genes that code for cell surface proteins essential for the acquired. e.g., adaptive immune system to recognize foreign molecules in vertebrates, which in turn determines histocompatibility. The MHC gene family is divided into three subgroups: MHC class I, MHC class II, and MHC class III. Class I MHC molecules have the β2 microglobulin subunit which can only be recognised by CD8 co-receptors. Class II MHC molecules have β1 and β2 subunits and can be recognized by CD4 co-receptors. In this way MHC molecules chaperone, which type of lymphocytes bind to the given antigen with high affinity, since different lymphocytes express different T-Cell Receptor (TCR) co-receptors. Components of the MHC are known in the art and can be readily identified by a skilled person. The MHC is further described in, e.g., Janeway C A Jr, Travers P, Walport M, et al, Immunobiology: The Immune System in Health and Disease, 5th edn (New York: Garland Science, 2001); Vigneron N, Stroobant V, Chapiro J, Ooms A, Degiovanni G, Morel S, et al. (April 2004). “An antigenic peptide produced by peptide splicing in the proteasome”. Science. 304 (5670): 587-90; and K. Murphy, “Antigen recognition by T cells,” in Janeway's Immunobiology, 8th, Ed., Garland Science, 2012, pp. 138-153; which are incorporated herein by reference in their entireties. A complete MHC class I complex comprises one MHC class I heavy chain, one peptide ligand sequence, and a beta 2 microglobulin. In some embodiments, a TCR recognition domain comprises one MHC class I heavy chain, one peptide ligand sequence, and a beta 2 microglobulin. A complete MHC class II complex comprises an MHC class II alpha chain, MHC class II beta chain, and one peptide ligand sequence. In some embodiments, a TCR recognition domain comprises an MHC class II alpha chain, MHC class II beta chain, and one peptide ligand sequence. These three components can either be assembled together as separate sequences (e.g., by intramolecular binding of multiple peptide molecules) or can be expressed as fusion proteins with intervening linker sequences (e.g., see Schmittnaegel et al., 2016; which is incorporated by reference herein in its entirety).

The MHC (Major Histocompatibility Complex), MHC-peptide complex, MHC-peptide fusion, featureless peptide MHC, can be selected on the basis of the disease or condition to be treated/prevented. Specific MHCs, peptides, and/or antigens that are associated with the diseases described herein are known in the art and an appropriate MHC, peptide, and/or antigen can be selected by one of ordinary skill in the art. For example, databases of suitable MHC, peptide, and/or antigen sequences are available on the world wide web at iedb.org; immunespace.org; immgen.org; import/org; peptideatlast.org/repository/; uniprot.org; ncbi.nlm.nih.gov/protein/; immunedata.org/index.php; immuneprofiling.org/hipc/; allergenonline.org/databasebrowe.shtml; and itntrialshare.org. Further examples are also provided in Smatti et al. 2019 Viruses 11:762; Beretta-Piccoli et al. 2019 J Autoimmu 94:1-6; and Cusick et al. 2012 Clinical Reviews in Allergy and Immunology; each of which is incorporated by reference herein in its entirety. The specific examples of TCR recognition domains provides herein are exemplary and non-limiting. One of skill in the art can identify relevant auto-antigenic pMHCs, allogeneic peptide MHCs, and autogenic peptide MHCs in addition to those described herein, e.g., from the art and/or from donor cells. Such identification is within the skill of the ordinary practitioner.

As an illustrative and non-limiting example, for compositions and methods relating to type 1 diabetes, the TCR recognition domain can comprise one or more of SEQ ID NOs: 8-17. In some embodiments, the TCR recognition domain can comprise SEQ ID NO: 8, 9, and one of 10-17. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to SEQ ID NO: 8, 9, and one of 10-17. In some embodiments, the TCR recognition domain can comprise sequences with at least 95% sequence identity to SEQ ID NO: 8, 9, and one of 10-17, and which retain the wild-type activity of SEQ ID NOs: 8, 9, and one of 10-17.

HLA-A*0201 (MHC class I heavy chain allele,

wildtype, human)

SEQ ID NO: 8

MAVMAPRTLVLLLSGALALTQTWAGSHSMRYFFTSVSRPGRGEPRFIAV

GYVDDTQFVRFDSDAASQRMEPRAPWIEQEGPEYWDGETRKVKAHSQTH

RVDLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRGYHQYAYDGKDY

TALKEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYL

ENGKETLQRTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDG

EDQTQDTELVETRPAGDGTFQKWAAVVVPSGQEQRYTCHVQHEGLPKPL

TLRWEPSSQPTIPIVGIIAGLVLFGAVITGAVVAAVMWRRKSSDRKGGS

YSQAASSDSAQGSDVSLTACKV

Beta-2 microglobulin (wildtype, human)

SEQ ID NO: 9

MSRSVALAVLALLSLSGLEGIQRTPKIQVYSRHPAENGKSNFLNCYVSG

FHQSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEY

ACRVNHVTLSQPKIVKWDRDM

Preproinsulin15-24 (wildtype, human)

SEQ ID NO: 10

ALWGPDPAAA

Preproinsulin15-24 altered peptide ligand #1

(synthetic, Cole et al., 2016)

SEQ ID NO: 11

AQWGPDPAAA

Preproinsulin15-24 altered peptide ligand #2

(synthetic, Cole et al., 2016)

SEQ ID NO: 12

RQWGPDPAAV

Preproinsulin15-24 altered peptide ligand #3

(wildtype, Clostridium asparagiforme, Cole

et al., 2016)

SEQ ID NO: 13

RQFGPDWIVA

Preproinsulin15-24 altered peptide ligand #4

(synthetic, Cole et al., 2016)

SEQ ID NO: 14

YQFGPDFPIA

Preproinsulin15-24 altered peptide ligand #5

(synthetic, Cole et al., 2016)

SEQ ID NO: 15

RQFGPDFPTI

Preproinsulin15-24 altered peptide ligand #6

(synthetic, Cole et al., 2016)

SEQ ID NO: 16

YLGGPDFPTI

Preproinsulin15-24 altered peptide ligand #7

(wildtype, Bacteroides fragilis, Cole et al.,

2016)

SEQ ID NO: 17

MVWGPDLYV

As a further illustrative example, for compositions and methods relating vitiligo, the TCR recognition domain can comprise one or more of SEQ ID NOs: 18-22. In some embodiments, the TCR recognition domain can comprise SEQ ID NO: 18, 19, and one of 20-22. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to SEQ ID NO: 18, 19, and one of 20-22. In some embodiments, the TCR recognition domain can comprise sequences with at least 95% sequence identity to SEQ ID NO: 18, 19, and one of 20-22, and which retain the wild-type activity of SEQ ID NOs: 18, 19, and one of 20-22.

HLA-A*0201 (MHC class I heavy chain allele,

wildtype, human, same as previous HLA-A2)

SEQ ID NO: 18

MAVMAPRTLVLLLSGALALTQTWAGSHSMRYFFTSVSRPGRGEPRFIAV

GYVDDTQFVRFDSDAASQRMEPRAPWIEQEGPEYWDGETRKVKAHSQTH

RVDLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRGYHQYAYDGKDY

TALKEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYL

ENGKETLQRTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDG

EDQTQDTELVETRPAGDGTFQKWAAVVVPSGQEQRYTCHVQHEGLPKPL

TLRWEPSSQPTIPIVGIIAGLVLFGAVITGAVVAAVMWRRKSSDRKGGS

YSQAASSDSAQGSDVSLTACKV

Beta-2 microglobulin (wildtype, human, same as

previous Beta-2 microglobulin)

SEQ ID NO: 19

MSRSVALAVLALLSLSGLEGIQRTPKIQVYSRHPAENGKSNFLNCYVSG

FHQSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEY

ACRVNHVTLSQPKIVKWDRDM

MART-126-35 (wildtype, human)

SEQ ID NO: 20

ELAGIGILTV

Tyrosinase368-376 (wildtype, human)

SEQ ID NO: 21

YMDGTMSQV

gp100209-217 (wildtype, human)

SEQ ID NO: 22

ITDQVPFSV

As further illustrative and non-limiting examples, the following pairs of MHC and antigens provided in Table 5 are known in the art. In some embodiments, the TCR recognition domain can comprise one or more of the following indicated MHC/peptide pairs, e.g., the TCR recognition domain can comprise one of the indicated MHC alleles and the indicated corresponding peptide. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of the following indicated MHC/peptide pairs, e.g., one of the indicated MHC alleles and the indicated corresponding peptide. In some embodiments, the TCR recognition domain can comprise sequences with at least 95% sequence identity to one of the following indicated MHC/peptide pairs, e.g., one of the indicated MHC alleles and the indicated corresponding peptide, wherein those sequences retain the wild-type activity of the MHC allele and the corresponding peptide.

TABLE 5
Each line of the following Table provides a MHC
allele (whose sequence is available in publically-
accessible databases, e.g., NCBI), and an antigen
sequence. The antigen source and/or the relevant
disease are also indicated on some lines. In some
examples provided in Table 5, the peptides are
antigen mimics and their origin is indicated.

HLA-A*0201 VMNILLQYVV (SEQ ID NO: 2013) GAD65

Diabetes

HLA-A*0201 VMNILLQYVV (SEQ ID NO: 2013) GAD65

Diabetes

HLA-A*0201 YAYDGKDYIA (SEQ ID NO: 2014) HLA-A2

Diabetes

HLA-A*0201 MVWESGCTV (SEQ ID NO: 2015) IA-2

Diabetes

HLA-A*0201 VIVMLTPLV (SEQ ID NO: 2016) IA-2

Diabetes

HLA-A*0201 YTCPLCRAPV (SEQ ID NO: 2017) SAA

Autoimmune

HLA-A*0201 YMDGTMSQV (SEQ ID NO: 2018) Tyrosinase

Vitiligo

HLA-A*0301 YMVPFIPLYR (SEQ ID NO: 2019) Tyrosinase

Vitiligo

HLA-A*2402 AFLPWHRLF (SEQ ID NO: 2020) Tyrosinase

Vitiligo

HLA-A*0101 SSDYVIPIGTY (SEQ ID NO: 2021) Tyrosinase

Vitiligo

HLA-A*02:01 VLHDDLLEA (SEQ ID NO: 2022) HA-1 137-

145 Minor Histocompatibility Antigen

HLA-A*02:01 RTLDKVLEV (SEQ ID NO: 2023) miHAg HA-8

Minor Histocompatibility Antigen

HLA-A*02:01 FIDSYICQV (SEQ ID NO: 2024) miHAg H-Y

(human SMCY) 311-319 Minor Histocompatibility

Antigen

DRB1*04:01 GAGSLQPLALEGSLQKRG A (SEQ ID NO: 2025)

Proinsulin 73-90

DRB1*04:01 IAFTSEHSHFSLK A (SEQ ID NO: 2026) GAD65

274-286

DRB1*15:01 DENPVVHFFKNIVTPRTPP (SEQ ID NO: 2027)

Myelin basic protein 83-101

DRB1*04:01 GIVEQCCTSICSLYQ A (SEQ ID NO: 2028)

Proinsulin 90-104

DRB1*04:01 NFIRMVISNPAAT A (SEQ ID NO: 2029) GAD65

555-567

DRB1*04:01 DVMNILLQYVVKSFDRSTKV (SEQ ID NO: 2030)

GAD65113-132

HLA-A*0201 ALWGPDPAAA (SEQ ID NO: 2031) Insulin

Diabetes

HLA-A*0201 HLVEALYLV (SEQ ID NO: 2032) Insulin

Diabetes

HLA-A*0201 KLQVFLIVL (SEQ ID NO: 2033) IAPP

Diabetes

HLA-A*0101 IVDCLTEMY (SEQ ID NO: 2034) USP9Y Graft

vs Host

HLA-A*0201 VLHDDLLEA (SEQ ID NO: 2035) HA-1 Graft

vs Host

HLA-A*0201 RTLDKVLEV (SEQ ID NO: 2036) HA-8 Graft

vs Host

HLA-A*0201 FIDSYICQV (SEQ ID NO: 2037) H-Y Graft

vs Host

HLA-B*0702 SPSVDKARAEL (SEQ ID NO: 2038) SMCY

Graft vs Host

HLA-B*0801 LPHNHTDL (SEQ ID NO: 2039) TPR-protein

Graft vs Host Antigen Mimicry

HLA-A*0201 FLDKGTYTL (SEQ ID NO: 2040) BALF4 EBV

HLA-A*0201 GLCTLVAML (SEQ ID NO: 2041) BMLF1 EBV

HLA-A*0201 TLDYKPLSV (SEQ ID NO: 2042) BMRF1 EBV

HLA-A*0201 YVLDHLIVV (SEQ ID NO: 2043) BRLF1 EBV

HLA-A*0201 LLDFVRFMGV (SEQ ID NO: 2044) EBNA 3B EBV

HLA-A*0201 YLLEMLWRL (SEQ ID NO: 2045) LMP-1 EBV

HLA-A*0201 YLQQNWWTL (SEQ ID NO: 2046) LMP-1 EBV

HLA-A*0201 CLGGLLTMV (SEQ ID NO: 2047) LMP-2A EBV

HLA-A*0201 FLYALALLL (SEQ ID NO: 2048) LMP-2A EBV

HLA-A*0301 RLRAEAQVK (SEQ ID NO: 2049) EMNA 3A EBV

HLA-A*1101 AVFDRKSDAK (SEQ ID NO: 2050) EBNA 3B EBV

HLA-A*1101 IVTDFSVIK (SEQ ID NO: 2051) EBNA 3B EBV

HLA-A*2402 DYCNVLNKEF (SEQ ID NO: 2052) BRLF1 EBV

HLA-A*2402 TYGPVFMCL (SEQ ID NO: 2053) LMP-2 EBV

HLA-A*2402 IACPIVMRY (SEQ ID NO: 2054) BRLF1 EBV

HLA-A*6801 IVTDFSVIK (SEQ ID NO: 2051) EBNA 3B EBV

HLA-B*0702 RPQGGSRPEFVKL (SEQ ID NO: 2055) BMRF1

EBF

HLA-B*0702 RPPIFIRRL (SEQ ID NO: 2056) EBNA 3A EBV

HLA-B*0702 QPRAPIRPI (SEQ ID NO: 2057) EBNA 6 EBV

HLA-B*0801 RAKFKQLL (SEQ ID NO: 2058) BZLF1 EBV

HLA-B*0801 FLRGRAYGL (SEQ ID NO: 2059) EBNA 3A EBV

HLA-B*3501 EPLPQGQLTAY (SEQ ID NO: 2060) BZLF1 EBV

HLA-B*3501 EPLSQSQITAY (SEQ ID NO: 2061) BZLF1 EBV

HLA-B*3501 HPVAEADYFEY (SEQ ID NO: 2062) EBNA 1 EBV

HLA-B*3501 HPVGDADYFEY (SEQ ID NO: 2063) EBNA 1 EBV

HLA-B*3501 HPVGEADYFEY (SEQ ID NO: 2064) EBNA 1 EBV

HLA-B*3501 HPVGQADYFEY (SEQ ID NO: 2065) EBNA 1 EBV

HLA-B*3501 YPLHEQHGM (SEQ ID NO: 2066) EBNA 3A EBV

HLA-A*0101 VTEHDTLLY (SEQ ID NO: 2067) UL44 CMV

HLA-A*0201 VLEETSVML (SEQ ID NO: 2068) IE-1 CMV

HLA-A*0201 NLVPMVATV (SEQ ID NO: 2069) pp65 CMV

HLA-A*0301 KLGGALQAK (SEQ ID NO: 2070) IE-1 CMV

HLA-A*2301 QYDPVAALF (SEQ ID NO: 2071) pp65 CMV

HLA-A*2402 AYAQKIFKI (SEQ ID NO: 2072) IE-1 CMV

HLA-A*2402 QYDPVAALF (SEQ ID NO: 2071) pp65 CMV

HLA-A*2402 VYALPLKML (SEQ ID NO: 2073) pp65 CMV

HLA-B*0702 RPHERNGFTVL (SEQ ID NO: 2074) pp65 CMV

HLA-B*0702 TPRVTGGGAM (SEQ ID NO: 2075) pp65 CMV

HLA-B*0801 ELKRKMIYM (SEQ ID NO: 2076) IE-1 CMV

HLA-B*0801 ELNRKMIYM (SEQ ID NO: 2077) IE-1 CMV

HLA-B*0801 ELRRKMMYM (SEQ ID NO: 2078) IE-1 CMV

HLA-B*0801 QIKVRVDMV (SEQ ID NO: 2079) IE-1 CMV

HLA-B*3501 IPSINVHHY (SEQ ID NO: 2080) pp65 CMV

HLA-B*3501 LPLNVGLPIIGVM (SEQ ID NO: 2081) UL138

CMV

A*01:01 SADNNNSEY (SEQ ID NO: 2082) AAV VP1 492-500

A*01:01 TDLGQNLLY (SEQ ID NO: 2083) Adenovirus 5

Hexon 886-894

A*01:01 EADPTGHSY (SEQ ID NO: 2084) MAGE-A1 161-169

A*01:01 EVDPIGHLY (SEQ ID NO: 2085) MAGE-A3 168-176

A*01:01 KSDICTDEY (SEQ ID NO: 2086) Tyrosinase

243-251 (244S)

A*01:01 KCDICTDEY (SEQ ID NO: 2087) Tyrosinase

243-251

A*01:01 QSLEIISRY (SEQ ID NO: 2088) Mcl-1 177-185

A*01:01 YVDFREYEYY (SEQ ID NO: 2089) FLT3 ITD

A*01:01 TLDTLTAFY (SEQ ID NO: 2090) Mesothelin

429-437

A*01:01 LTDDRLFTCY (SEQ ID NO: 2091) PLEKHM2

A*01:01 DSDPDSFQDY (SEQ ID NO: 2092) Tyr A1a

454-463

A*01:01 EADPIGHLY (SEQ ID NO: 2093) MAGEA3

A*01:01 EVDPASNTY (SEQ ID NO: 2094) MAGE-A4 169-177

A*01:01 HSTNGVTRIY (SEQ ID NO: 2095) PSMA

A*01:01 ILDTAGREEY (SEQ ID NO: 2096) N-ras 55-64

A*01:01 LVDVMPWLQY (SEQ ID NO: 2097) Cytochrome

P450 240-249

A*01:01 RSDSGQQARY (SEQ ID NO: 2098) AIM-2

A*01:01 VTEPGTAQY (SEQ ID NO: 2099) Minor antigen

HA-3T (Lbc oncogene 451-459)

A*01:01 VYDFFVWLHY (SEQ ID NO: 2100) TRP-2 181-190

A*01:01 YSEHPTFTSQY (SEQ ID NO: 2101) HCMV pp50

245-253

A*01:01 VTEHDTLLY (SEQ ID NO: 2102) HCMV pp50

245-253

A*01:01 FTSDYYQLY (SEQ ID NO: 2103) SARS-CoV-2

ORF3a 207-215 (confirmed epitope)

A*01:01 TTDPSFLGRY (SEQ ID NO: 2104) SARS-CoV-2

Replicase polyprotein 1ab 1637-1646

A*01:01 PTDNYITTY (SEQ ID NO: 2105) SARS-CoV-2

Replicase polyprotein 1ab 1621-1629

A*01:01 LLDTASALY (SEQ ID NO: 2106) HBV core 30-38

A*01:01 ATDALMTGY (SEQ ID NO: 2107) HCV NS3

1435-1443

A*01:01 ATDALMTGF (SEQ ID NO: 2108) HCV NS3

1436-1444

A*01:01 CTELKLSDY (SEQ ID NO: 2109) Influenza A

(PR8) NP 44-52

A*01:01 VSDGGPNLY (SEQ ID NO: 2110) Influenza A

PB1 591-599

A*01:01 IVDCLTEMY (SEQ ID NO: 2111) DRRFYY

(1521-1529))

A*0201 ALCNTDSPL (SEQ ID NO: 2112) iLR1

A*0201 ALKDVEERV (SEQ ID NO: 2113) MAGE-C2 336-344

A*0201 LLAARAIVAI (SEQ ID NO: 2114) iLR1 59-68

A*0201 RLWQELSDI (SEQ ID NO: 2115) circadian clock

protein PASD1 691-700

A*0201 LLFGLALIEV (SEQ ID NO: 2116) MAGE-C2 191-200

A*0201 FLDPRPLTV (SEQ ID NO: 2117) CYP190

A*0201 STLCQVEPV (SEQ ID NO: 2118) MPP11

A*0201 VLQMKEEDV (SEQ ID NO: 2119) iLR1

A*0201 AIQDLCLAV (SEQ ID NO: 2120) NPM1

A*0201 QLLIKAVNL (SEQ ID NO: 2121) MPP11

A*0201 AIQDLCVAV (SEQ ID NO: 2122) NPM1

A*0201 ALTPVVVTL (SEQ ID NO: 2123) cyclin-dependent

kinase 4 170-178

A*02:01 KLQVFLIVL (SEQ ID NO: 2124) T1D Diabetes

human prepro islet amyloid polypeptide pplAPP 513

A*02:01 VMNILLQYV (SEQ ID NO: 2125) GAD65 114-123

A*02:01 SLSRFSWGA (SEQ ID NO: 2126) Myelin basic

protein 110-118

A*02:01 HLVEALYLV (SEQ ID NO: 2127) Insulin B

chain 10-18

A*02:01 LNIDLLWSV (SEQ ID NO: 2128) T1D Diabetes

IGRP 228-236

A*02:01 VLFGLGFAI (SEQ ID NO: 2129) T1D Diabetes

IGRP 265-273

A*02:01 ALWGPDPAAA (SEQ ID NO: 2130) Proinsulin

precursor 15-24

A*02:01 MVWESGCTV (SEQ ID NO: 2131) IA-2 797-805

A*02:01 YTCPLCRAPV (SEQ ID NO: 2132) SSA SS-56

55-64

A*02:01 VIVMLTPLV (SEQ ID NO: 2133) IA-2 805-813

A*02:01 AITEVECFL (SEQ ID NO: 2134) VP1 44-52

A*02:01 FLHCIVFNV (SEQ ID NO: 2135) large T

antigen 410-418

A*02:01 LLMWEAVTV (SEQ ID NO: 2136) VP1 108-116

A*02:01 CLLPKMDSV (SEQ ID NO: 2137) large T

antigen 398-406

A*02:01 FLWGPRALV (SEQ ID NO: 2138) MAGEA3 271-279

A*02:01 IMDQVPFSV (SEQ ID NO: 2139) gp100 (pmel17)

209-217

A*02:01 YLEPGPVTV (SEQ ID NO: 2140) gp100 (pmel)

280-288 (288V)

A*02:01 YLSGADLNL (SEQ ID NO: 2141)

Carcinoembryonic antigen (CEA)-derived peptide

CAP1-6D

A*02:01 SLLMWITQC (SEQ ID NO: 2142) NY-ESO-1

157-165 (9C)

A*02:01 KTWGQYWQV (SEQ ID NO: 2143) gp100 (pmel17)

154-162

A*02:01 YLEPGPVTA (SEQ ID NO: 2144) gp100

A*02:01 YMDGTMSQV (SEQ ID NO: 2145) Tyrosinase

369-377 (371D)

A*02:01 YLSGANLNL (SEQ ID NO: 2146) Carcinogenic

Embryonic Antigen (CEA) 571-579

A*02:01 ELAGIGILTV (SEQ ID NO: 2147) MelanA/MART

26-35

A*02:01 ILAKFLHWL (SEQ ID NO: 2148) Telomerase

540-548

A*02:01 ALQPGTALL (SEQ ID NO: 2149) Prostate Stem

Cell Antigen (PSCA) 14-22

A*02:01 VISNDVCAQV (SEQ ID NO: 2150)Prostate

Specific Antigen-1 (PSA-1) 154-163

A*02:01 RLVDDFLLV (SEQ ID NO: 2151) Telomerase

Reverse Transcriptase 865-873

A*02:01 GVLVGVALI (SEQ ID NO: 2152) Carcinogenic

Embryonic Antigen (CEA) 694-702

A*02:01 VLYRYGSFSV (SEQ ID NO: 2153) gp100

(pmel17) 476-485

A*02:01 PLFQVPEPV (SEQ ID NO: 2154) Alpha-

fetoprotein isoform 1 137-145

A*02:01 FMNKFIYEI (SEQ ID NO: 2155) Human alfa

fetoprotein 158-166

A*02:01 GLSPNLNRFL (SEQ ID NO: 2156) Alpha-

fetoprotein isoform 2 167-176

A*02:01 KVLEYVIKV (SEQ ID NO: 2157) MAGEA1 278-286

A*02:01 LLGRNSFEV (SEQ ID NO: 2158) p53 264-272

A*02:01 LLLLTVLTV (SEQ ID NO: 2159) MUC-1 12-20

A*02:01 ILHNGAYSL (SEQ ID NO: 2160) HER-2/neu

435-443

A*02:01 RLLQETELV (SEQ ID NO: 2161) HER-2/neu

689-697

A*02:01 KIFGSLAFL (SEQ ID NO: 2162) HER-2/neu

369-377

A*02:01 LLLLDVAPL (SEQ ID NO: 2163) HSP1A 459-467

A*02:01 LLDVAPLSL (SEQ ID NO: 2164) HSP1A 461-469

A*02:01 HLYQGCQVV (SEQ ID NO: 2165) Receptor

tyrosine-protein kinase erbB-2 48-56

A*02:01 HLSTAFARV (SEQ ID NO: 2166) G250 (renal

cell carcinoma) 217-225

A*02:01 VLQELNVTV (SEQ ID NO: 2167) Leukocyte

Proteinase-3 (Wegener's autoantigen) 169-177

A*02:01 KVAELVHFL (SEQ ID NO: 2168) MAGEA3 112-120

A*02:01 VLAGVGFFI (SEQ ID NO: 2169) EPHA2 550-558

A*02:01 FLYTLLREV (SEQ ID NO: 2170) STEAP 86-94

A*02:01 ILLWQPIPV (SEQ ID NO: 2171) Prostatic Acid

Phosphatase-3 (PAP-3) 135-143

A*02:01 RLQEERTCKV (SEQ ID NO: 2172) BIR

A*02:01 QLCPICRAPV (SEQ ID NO: 2173) Livin/ML-

IAP280 175-184

A*02:01 VLGEAWRDQV (SEQ ID NO: 2174) TRAP 45-54

A*02:01 LLLTVLTVV (SEQ ID NO: 2175) Tumor Mucin

Antigen 13-21

A*02:01 GLYDGMEHL (SEQ ID NO: 2176) MAGEA-10

254-262

A*02:01 SLLMWITQV (SEQ ID NO: 2177) NY-ESO-1

157-165

A*02:01 LMLGEFLKL (SEQ ID NO: 2178) Survivin 96-104

A*02:01 YLFFYRKSV (SEQ ID NO: 2179) mTERT 572-580

A*02:01 ELTLGEFLKL (SEQ ID NO: 2180) survivin

95-104

A*02:01 FLTPKKLQCV (SEQ ID NO: 2181) Prostate

Specific Antigen-1 (PSA-1) 141-150

A*02:01 KLQCVDLHV (SEQ ID NO: 2182) Prostate

Specific Antigen 146-154

A*02:01 TLAPATEPA (SEQ ID NO: 2183) Mucin 79-87

A*02:01 YLQVNSLQTV (SEQ ID NO: 2184) Telomerase

Reverse Transcriptase (hTRT) 988-997

A*02:01 SLGEQQYSV (SEQ ID NO: 2185) WT1 187-195

A*02:01 SLEENIVIL (SEQ ID NO: 2186) RHAMM 275-283

A*02:01 YMNGTMSQV (SEQ ID NO: 2187) Tyrosinase

368-376

A*02:01 ILSLELMKL (SEQ ID NO: 2188) Receptor for

hyaluronic acid-mediatedmotility (RHAMM) 165-173

A*02:01 PLFDFSWLSL (SEQ ID NO: 2189) Bcl-2 208-217

A*02:01 LLGATCMFV (SEQ ID NO: 2190) CyclinD 101-109

A*02:01 ALYVDSLFFL (SEQ ID NO: 2191) PRAME PRA

300-309

A*02:01 GLMEEMSAL (SEQ ID NO: 2192) Human Mena

protein (overexpressed in breast cancer)

A*02:01 TMNGSKSPV (SEQ ID NO: 2193) hMena 502-510

A*02:01 GVYDGREHTV (SEQ ID NO: 2194) MAGE-A4

230-239

A*02:01 YLNDHLEPWI (SEQ ID NO: 2195) Bcl-X 173-182

A*02:01 ALDVYNGLL (SEQ ID NO: 2196) Prostatic acid

phosphatase precursor (PAP) 299-307

A*02:01 ALFDIESKV (SEQ ID NO: 2197) PSM P2

(prostate)

A*02:01 SLAMLDLLHV (SEQ ID NO: 2198) Mutant

anaplastic lymphoma kinase 1220-1229

A*02:01 YLNTVQPTCV (SEQ ID NO: 2199) EGF-R

1138-1147

A*02:01 KLFGTSGQKT (SEQ ID NO: 2200) EGF-R-479

350-359

A*02:01 RMPEAAPPV (SEQ ID NO: 2201) p53 65-73

A*02:01 PLTSIISAV (SEQ ID NO: 2202) Receptor

tyrosine-protein kinase erbB-2 728-736

A*02:01 VLAGGFFLL (SEQ ID NO: 2203) PSMA 27-38

A*02:01 LLHETDSAV (SEQ ID NO: 2204) PSMA/PSM-P1

4-12

A*02:01 VMAGVGSPYV (SEQ ID NO: 2205) Receptor

tyrosine-protein kinase erbB-2 819-828

A*02:01 VLPLTVAEV (SEQ ID NO: 2206) Mesothelin

530-538

A*02:01 SLLFLLFSL (SEQ ID NO: 2207) Mesothelin

20-28

A*02:01 QLFEELQEL (SEQ ID NO: 2208) Heme oxygenase-

1 212-220

A*02:01 VLDGLDVLL (SEQ ID NO: 2209) PRAME 100-108

A*02:01 RLASFYDWPL (SEQ ID NO: 2210) BIR7 90-99

A*02:01 LIAHNQVRQV (SEQ ID NO: 2211) HER-2/neu

(85-94)

A*02:01 ILHDGAYSL (SEQ ID NO: 2212) HER-2 434-443

A*02:01 FVGEFFTDV (SEQ ID NO: 2213) GPC3 144-152

(overexpressed in hepatocellular carcinoma)

A*02:01 LLLIWFRPV (SEQ ID NO: 2214) BKV Ltag

579-587

A*02:01 KLQDASAEV (SEQ ID NO: 2215) HM1.24-aa

126-134

A*02:01 SLYSFPEPEA (SEQ ID NO: 2216) PRAME

A*02:01 SLLQHLIGL (SEQ ID NO: 2217) PRAME 425-433

A*02:01 VIFDFLHCI (SEQ ID NO: 2218) BKV Ltag

406-414

A*02:01 VLDFAPPGA (SEQ ID NO: 2219) WT1

A*02:01 TLPGYPPHV (SEQ ID NO: 2220) PAX-5 311-319

A*02:01 YMEHNNVYTV (SEQ ID NO: 2221) Fibromodulin

250-259

A*02:01 YLQHNEIQEV (SEQ ID NO: 2222) Fibromodulin

206-215

A*02:01 SLVDVMPWL (SEQ ID NO: 2223) Cytochrome

p450 1B1 239-248

A*02:01 RLMNDMTAV (SEQ ID NO: 2224) HSP105 128-136

A*02:01 RLARLALVL (SEQ ID NO: 2225) Trophoblast

glycoprotein 17-25

A*02:01 FLTGNQLAV (SEQ ID NO: 2226) 5T4 97-105

A*02:01 LLLAGLFSL (SEQ ID NO: 2227) Fibromodulin

7-15

A*02:01 FLGYLILGV (SEQ ID NO: 2228) Prostatic Acid

Phosphatase-3 (PAP-3)

A*02:01 SLFLGILSV (SEQ ID NO: 2229) CD20 188-196

(B cell malignancies)

A*02:01 AVLPLLELV (SEQ ID NO: 2230) MCL-1 139-147

A*02:01 SLSEKTVLL (SEQ ID NO: 2231) CD59

glycoprotein precursor 106-114

A*02:01 YMCSFLFNL (SEQ ID NO: 2232) Ewing Tumor

EZH2 666-674

A*02:01 YLISGDSPV (SEQ ID NO: 2233) CD33 65-73

(1Y2L)

A*02:01 KASEKIFYV (SEQ ID NO: 2234) SSX2 41-49

A*02:01 FLAKLNNTV (SEQ ID NO: 2235) HCA587 317-325

A*02:01 GLAPPQHLIRV (SEQ ID NO: 2236) p53 187-197

A*02:01 VIMPCSWWV (SEQ ID NO: 2237) Chondromodulin-

I 319-327

A*02:01 KVVEFLAML (SEQ ID NO: 2238) MAGE-C1

1083-1091

A*02:01 LTLGEFLKL (SEQ ID NO: 2239) Survivin-3A

96-104

A*02:01 ALPFGFILV (SEQ ID NO: 2240) IL13R 345-353

A*02:01 TLADFDPRV (SEQ ID NO: 2241) EphA2

A*02:01 ALMEQQHYV (SEQ ID NO: 2242) ITGB8 662-670

A*02:01 CLTSTVQLV (SEQ ID NO: 2243) HER-2/neu

789-797

A*02:01 GLLGASVLGL (SEQ ID NO: 2244) Telomerase

Reverse Transcriptase (hTRT) 674-683

A*02:01 QLLDGFMITL (SEQ ID NO: 2245) PASD1 39-48

A*02:01 YLVGNVCIL (SEQ ID NO: 2246) PASD1 168-176

A*02:01 ALLTSRLRFI (SEQ ID NO: 2247) Telomerase

Reverse Transcriptase (hTRT) 615-624

A*02:01 RLSSCVPVA (SEQ ID NO: 2248) TGF beta

receptor type-2 131-139

A*02:01 FLYDDNQRV (SEQ ID NO: 2249) Topoisomerase

II-alpha-b 828-836

A*02:01 YLIELIDRV (SEQ ID NO: 2250) TACE 250-258

A*02:01 FLAEDALNTV (SEQ ID NO: 2251) Epithelial

Discoidin Domain Receptor 1 (EDDR1) 867-876

A*02:01 GLMKYIGEV (SEQ ID NO: 2252) TRPM8 187-195

A*02:01 AILALLPAL (SEQ ID NO: 2253) Prostate Stem

Cell Antigen (PSCA) 105-133

A*02:01 GLQHWVPEL (SEQ ID NO: 2254) BA46

(Lactadherin) 97-106

A*02:01 GVRGRVEEI (SEQ ID NO: 2255) BCR-ABL

A*02:01 ITDQVPFSV (SEQ ID NO: 2256) gp100 (pmel)

209-217

A*02:01 KLCPVQLWV (SEQ ID NO: 2257) p53 139-147

A*02:01 KVAEELVHFL (SEQ ID NO: 2258) MAGEA3 112-120

(alternative version)

A*02:01 SLPPPGTRV (SEQ ID NO: 2259) p53 149-157

A*02:01 YLGSYGFRL (SEQ ID NO: 2260) p53 103-111

A*02:01 YLQLVFGIEV (SEQ ID NO: 2261) MAGEA2 157-166

A*02:01 TLQDIVYKL (SEQ ID NO: 2262) BMI1 74-82

A*02:01 YAIDLPVSV (SEQ ID NO: 2263) L-dopachrome

tautomerase 488-496

A*02:01 AMVGAVLTA (SEQ ID NO: 2264) Tyrosinase

482-190

A*02:01 ATVGIMIGV (SEQ ID NO: 2265) CEACAM5 687-695

A*02:01 YVDPVITSI (SEQ ID NO: 2266) Hepatocyte

growth factor receptor 673-681

A*02:01 GVLLWEIFSL (SEQ ID NO: 2267) VEGFR1 28-37

A*02:01 LMAQEALAFL (SEQ ID NO: 2268) CAMEL 2-11

A*02:01 RVA(PHOSPHO-S)PTSGV (SEQ ID NO: 2269)

Insulin receptor substrate-2 1097-1105

A*02:01 RVASPTSGV (SEQ ID NO: 2270) IRS-2 1097-1105

A*02:01 ALNVYNGLL (SEQ ID NO: 2271) ACPP 299-307

A*02:01 ALSPVPPVV (SEQ ID NO: 2272) Bcl-2 85-93

A*02:01 ALVCYGPGI (SEQ ID NO: 2273) FAP alpha

463-471

A*02:01 ALWPWLLMAT (SEQ ID NO: 2274) RNF43 11-20

A*02:01 ALYLMELTM (SEQ ID NO: 2275) CB9L2

A*02:01 CLPSPSTPV (SEQ ID NO: 2276) BMI1 271-279

A*02:01 ELSDSLGPV (SEQ ID NO: 2277) PASD1 695-703

A*02:01 FLFLRNFSL (SEQ ID NO: 2278) TARP(V28L)

27-35

A*02:01 FLPSPLFFFL (SEQ ID NO: 2279) TARP(P5L) 5-13

A*02:01 GLFKCGIAV (SEQ ID NO: 2280) FAP 639-647

A*02:01 GLIQLVEGV (SEQ ID NO: 2281) TRAG-3 4-12

A*02:01 ILGVLTSLV (SEQ ID NO: 2282) DLK1 309-317

A*02:01 LLVPTCVFLV (SEQ ID NO: 2283) 691-700

A*02:01 MLAVFLPIV (SEQ ID NO: 2284) STEAP 292-300

(293L)

A*02:01 NLFETPVEA (SEQ ID NO: 2285) 194-202

A*02:01 QLGEQCWTV (SEQ ID NO: 2286) PSCA 44-51

(51A)

A*02:01 RLAEYQAYI (SEQ ID NO: 2287) SART3 309-317

A*02:01 SIDWFMVTV (SEQ ID NO: 2288) p31-39

A*02:01 SILLRDAGLV (SEQ ID NO: 2289) TRAG-3 57-66

A*02:01 SLFEPPPPG (SEQ ID NO: 2290) PSMA 85-93

A*02:01 SQADALKYV (SEQ ID NO: 2291) EZH2 729-737

A*02:01 WLSLKTLLSL (SEQ ID NO: 2292) Bcl-2 214-223

A*02:01 YLNRHLHTWI (SEQ ID NO: 2293) BCL-2 180-189

A*02:01 YLQWIEFSI (SEQ ID NO: 2294) Prominin1

744-752

A*02:01 YLYQWLGAPV (SEQ ID NO: 2295) Osteocalcin

51-60

A*02:01 KLMSSNSTDL (SEQ ID NO: 2296) HSP105 234-243

A*02:01 RLQGISPKI (SEQ ID NO: 2297) SSX2 103-111

A*02:01 AILALLPALL (SEQ ID NO: 2298) PSCA

A*02:01 ALIHHNTHL (SEQ ID NO: 2299) HER2 466-474

A*02:01 CMHLLLEAV (SEQ ID NO: 2300) MG50 624-632

A*02:01 FLIIWQNTM (SEQ ID NO: 2301) FSP26

A*02:01 FLPWHRLFLL (SEQ ID NO: 2302) Tyrosinase

207-216

A*02:01 FVWLHYYSV (SEQ ID NO: 2303) TRP2 185-193(L)

A*02:01 GLFGDIYLA (SEQ ID NO: 2304) CSNK1A1 26-34

A*02:01 GLFGDIYLAI (SEQ ID NO: 2305) CSNK1A1 26-35

A*02:01 ILLRDAGLV (SEQ ID NO: 2306) TRAG-3L 58-66

A*02:01 ILLVVVLGV (SEQ ID NO: 2307) Receptor

tyrosine-protein kinase erbB-2 707-715

A*02:01 ILNAMIAKI (SEQ ID NO: 2308) HAUS3 154-162

A*02:01 KASEYLQLV (SEQ ID NO: 2309) MAGEA2 153-161

A*02:01 KIWEELSVL (SEQ ID NO: 2310) MAGEA3 220-228

A*02:01 KLIDRTE(S)L (SEQ ID NO: 2311) LSP1 325-333

A*02:01 KLTGDENFTI (SEQ ID NO: 2312) Tyrosinase

precursor 224-233

A*02:01 LLCYSCKAQV (SEQ ID NO: 2313) PSCA 17-26

A*02:01 LLLEAVPAV (SEQ ID NO: 2314) MG50 69-77

A*02:01 LLNQLQVNL (SEQ ID NO: 2315) Mucin2 467-475

A*02:01 LLRDAGLVKM (SEQ ID NO: 2316) TRAP 59-68

A*02:01 LLRRYNVAKV (SEQ ID NO: 2317) SOX11 266-275

A*02:01 LLSHGAVIEV (SEQ ID NO: 2318) Ankyrin NYBR1

158-167

A*02:01 LVFGIELMEV (SEQ ID NO: 2319) MAGEA3 160-169

A*02:01 LVFGIEVVEV (SEQ ID NO: 2320) MAGEA12

160-169

A*02:01 MLWGWREHV (SEQ ID NO: 2321) Mucin2 645-653

A*02:01 PLQPEQLQV (SEQ ID NO: 2322) Receptor

tyrosine-protein kinase erbB-2 437-445

A*02:01 QLMAFNHLI (SEQ ID NO: 2323) PAX3/FKHR

135-143

A*02:01 QLMPYGCLL (SEQ ID NO: 2324) Receptor

tyrosine-protein kinase erbB-2 845-853

A*02:01 RLGPTLMCL (SEQ ID NO: 2325) MG50 1244-1252

A*02:01 RLTRFLSRV (SEQ ID NO: 2326) CyclinD 228-236

A*02:01 RTF(S)PTYGL (SEQ ID NO: 2327) Desmuslin

426-434

A*02:01 SILLRDAGL (SEQ ID NO: 2328) TRAP 57-65

A*02:01 SLADEAEVYL (SEQ ID NO: 2329) GAS7

Neoepitope

A*02:01 SLDDYNHLV (SEQ ID NO: 2330) L-dopachrome

tautomerase 288-296

A*02:01 SLYKFSPFPL (SEQ ID NO: 2331) O-linked N-

acetylglucosamine transferase FSP06

A*02:01 SMTR(S)PPRV (SEQ ID NO: 2332) SFRS2B

241-249

A*02:01 TLEEITGYL (SEQ ID NO: 2333) Receptor

tyrosine-protein kinase erbB-2 448-456

A*02:01 TLHCDCEIL (SEQ ID NO: 2334) MG50 210-218

A*02:01 VLEPPGARDV (SEQ ID NO: 2335) BIR 7 230-239

A*02:01 VLLALLMAGL (SEQ ID NO: 2336) Prostate stem

cell antigen 4-13

A*02:01 VLSVNVPDV (SEQ ID NO: 2337) MG50 625-633

A*02:01 VLVKSPNHV (SEQ ID NO: 2338) Receptor

tyrosine-protein kinase erbB-4 890-898

A*02:01 VMIG(S)PKKV (SEQ ID NO: 2339) Tensin3

1558-1566

A*02:01 VVLGVVFGI (SEQ ID NO: 2340) Receptor

tyrosine-protein kinase erbB-2 743-751

A*02:01 WLPKILGEV (SEQ ID NO: 2341) MG50 1051-1059

A*02:01 WLQYFPNPV (SEQ ID NO: 2342) Cytochrome P450

246-254

A*02:01 YLLDLSTNHL (SEQ ID NO: 2343) Fibromodulin

7-15

A*02:01 YLWWVNNQSL (SEQ ID NO: 2344) CEA 176-185

A*02:01 ALGGHPLLGV (SEQ ID NO: 2345) Dickkopf-

related protein 1 20-29

A*02:01 ALLAGLVSL (SEQ ID NO: 2346) FGFR4 676-684

A*02:01 ALLTYMIAHI (SEQ ID NO: 2347) Thymidylate

synthase 231-240

A*02:01 ALMDKSLHV (SEQ ID NO: 2348) MART-1 56-64

A*02:01 ALPPPLMLL (SEQ ID NO: 2349) Heparanase 8-16

A*02:01 ALSVMGVYV (SEQ ID NO: 2350) MAGEA9 223-231

A*02:01 ALVEFEDVL (SEQ ID NO: 2351) hnRNP L 140-148

A*02:01 ALWPWLLMA (SEQ ID NO: 2352) RNF43 11-19

A*02:01 AMLGTHTMEV (SEQ ID NO: 2353) Melanocyte-

specific secreted glycoprotein 184-193

A*02:01 AVIGALLAV (SEQ ID NO: 2354) Melanocyte-

specific secreted glycoprotein 20-28

A*02:01 CLYGNVEKV (SEQ ID NO: 2355) hnRNP L 404-412

A*02:01 DLIFGLNAL (SEQ ID NO: 2356) Heparanase

185-193

A*02:01 ELFQDLSQL (SEQ ID NO: 2357) ETV5 54-53

A*02:01 FAWERVRGL (SEQ ID NO: 2358) Cyclin-

dependent kinase inhibitor 1 97-105

A*02:01 FIASNGVKLV (SEQ ID NO: 2359) ACTN4 118-127

(K5N)

A*02:01 FLALIICNA (SEQ ID NO: 2360) Tubulin beta 4

283-291

A*02:01 FLDEFMEGV (SEQ ID NO: 2361) Malic enzyme

224-232

A*02:01 RMFPNAPYL (SEQ ID NO: 2362) WT-1 126-134

(Wilms tumor)

A*02:01 RLNMFTPYI (SEQ ID NO: 2363) Chlamydia

trachomatis MOMP 258-266

A*02:01 NMFTPYIGV (SEQ ID NO: 2364) MOMP precursor

283-291

A*02:01 NLVPMVATV (SEQ ID NO: 2365) HCMV pp65

495-504

A*02:01 VLEETSVML (SEQ ID NO: 2366) HCMV IE1

316-324 (UL123)

A*02:01 VLAELVKQI (SEQ ID NO: 2367) HCMV IE1 81-89

A*02:01 MLNIPSINV (SEQ ID NO: 2368) pp65 120-128

A*02:01 LLLDRLNQL (SEQ ID NO: 2369) SARS-CoV

Nucleocapsid protein 223-231 (conserved inSARS-

CoV-2)

A*02:01 FIAGLIAIV (SEQ ID NO: 2370) SARS-CoV-2

Spike glycoprotein 1220-1228 (confirmed epitope)

A*02:01 ALNTLVKQL (SEQ ID NO: 2371) SARS-CoV Spike

glycoprotein precursor 940-948 (conserved in SARS-

CoV-2)

A*02:01 LITGRLQSL (SEQ ID NO: 2372) SARS-CoV-2

Spike glycoprotein 996-1004 (confirmed epitope)

A*02:01 NLNESLIDL (SEQ ID NO: 2373) SARS-CoV

Spike glycoprotein precursor 1174-1182 (conserved

in SARS-Cov-2)

A*02:01 VLNDILSRL (SEQ ID NO: 2374) SARS-CoV Spike

glycoprotein precursor 958-966 (conserved in SARS-

Cov-2)

A*02:01 YLQPRTFLL (SEQ ID NO: 2375) SARS-Cov-2

Spike glycoprotein 269-277 (confirmed epitope)

A*02:01 LLYDANYFL (SEQ ID NO: 2376) SARS-CoV-2

ORF3a 139-147 (confirmed epitope)

A*02:01 RLQSLQTYV (SEQ ID NO: 2377) SARS-CoV-2

Spike glycoprotein 1000-1008 (confirmed

subdominant epitope)

A*02:01 KLWAQCVQL (SEQ ID NO: 2378) SARS-CoV-2

ORF1ab 3886-3894 (confirmed epitope)

A*02:01 TLYAVATTI (SEQ ID NO: 2379) Dengue NS4b

40-48

A*02:01 KLAEAIFKL (SEQ ID NO: 2380) Dengue NS5

563-571

A*02:01 ILIRTGLLVI (SEQ ID NO: 2381) Dengue NS2b

97-106

A*02:01 AIKRGLRTL (SEQ ID NO: 2382) Dengue NS3

112-120

A*02:01 LLLGLMILL (SEQ ID NO: 2383) Dengue NS4a

56-64

A*02:01 VLLLVTHYA (SEQ ID NO: 2384) Dengue NS4b

111-119

A*02:01 GLCTLVAML (SEQ ID NO: 2385) EBV BMLF-1

259-267

A*02:01 CLGGLLTMV (SEQ ID NO: 2386) EBV LMP-2

426-434

A*02:01 YLLEMLWRL (SEQ ID NO: 2387) EBV LMP-1

125-133

A*02:01 YLQQNWWTL (SEQ ID NO: 2388) EBV LMP1

159-167

A*02:01 YVLDHLIVV (SEQ ID NO: 2389) EBV BRLF1

109-117

A*02:01 FLYALALLL (SEQ ID NO: 2390) EBV LMP-2

356-364

A*02:01 TLDYKPLSV (SEQ ID NO: 2391) EBV BMRF1

208-216

A*02:01 LLDFVRFMGV (SEQ ID NO: 2392) EBV EBNA-3C

284-293

A*02:01 FLDKGTYTL (SEQ ID NO: 2393) EBV BALF-4

276-284

A*02:01 FLPSDFFPSV (SEQ ID NO: 2394) HBV core

antigen 18-27

A*02:01 FLLTRILTI (SEQ ID NO: 2395) HBV envelope

183-191

A*02:01 GLSPTVWLSV (SEQ ID NO: 2396) HBV surface

antigen 185-194

A*02:01 WLSLLVPFV (SEQ ID NO: 2397) HBV surface

antigen 172-181

A*02:01 FLLSLGIHL (SEQ ID NO: 2398) HBV polymerase

573-581

A*02:01 FLPSDFFPSI (SEQ ID NO: 2399) HBV core

18-27 (subtype ADR4)

A*02:01 VLHKRTLGL (SEQ ID NO: 2400) HBV X 92-100

A*02:01 GLSRYVARL (SEQ ID NO: 2401) HBV Pol 455-463

A*02:01 YMDDVVLGA (SEQ ID NO: 2402) HBV Polymerase

548-556

A*02:01 KLHLYSHPI (SEQ ID NO: 2403) HBV Pol 502-510

A*02:01 ELMTLATWV (SEQ ID NO: 2404) HBV core

protein 64-72

A*02:01 DLMGYIPAV (SEQ ID NO: 2405) HCV core

132-140

A*02:01 CINGVCWTV (SEQ ID NO: 2406) HCV NS3

1073-1081

A*02:01 YLLPRRGPRL (SEQ ID NO: 2407) HCV core 35-44

A*02:01 VLSDFKTWL (SEQ ID NO: 2408) HCV NS5a

1987-1995

A*02:01 ALYDVVTKL (SEQ ID NO: 2409) HCV NS5b

2594-2602

A*02:01 KLVALGINAV (SEQ ID NO: 2410) HCV NS3

1406-1415

A*02:01 LLFNILGGWV (SEQ ID NO: 2411) HCV NS4b

1807-1816

A*02:01 KLSGLGINAV (SEQ ID NO: 2412) HCV NS3

1406-1415

A*02:01 DLMGYIPLV (SEQ ID NO: 2413) HCV core

132-140

A*02:01 CVNGVCWTV (SEQ ID NO: 2414) HCV NS3

1073-1081

A*02:01 GLQDCTMLV (SEQ ID NO: 2415) HCV NS5B

2727-2735

A*02:01 SLYNTVATL (SEQ ID NO: 2416) HIV-1 gag p17

76-84

A*02:01 ILKEPVHGV (SEQ ID NO: 2417) HIV-1 RT

476-484

A*02:01 TLNAWVKVV (SEQ ID NO: 2418) HIV-1 gag p24

19-27

A*02:01 KLTPLCVTL (SEQ ID NO: 2419) HIV-1 env

gp120 90-98

A*02:01 GLADQLIHL (SEQ ID NO: 2420) HIV-1 vif

101-109

A*02:01 LTFGWCFKL (SEQ ID NO: 2421) HIV-1 nef

137-145

A*02:01 FLGKIWPS (SEQ ID NO: 2422) Gag 433-440

A*02:01 ALVEMGHHA (SEQ ID NO: 2423) HIV Vpu 66-74

A*02:01 RTLNAWVKV (SEQ ID NO: 2424) HIV gag 150-158

A*02:01 NVWATHACV (SEQ ID NO: 2425) HIV env gp 67-7

A*02:01 SLLNATAIAV (SEQ ID NO: 2426) HIV env

816-825

A*02:01 SLFNTVATL (SEQ ID NO: 2427) HIV gag 77-85

A*02:01 SLVKHHMYI (SEQ ID NO: 2428) HIV vif 23-31

A*02:01 VIYHYVDDL (SEQ ID NO: 2429) HIV pol

A*02:01 YMLDLQPETT (SEQ ID NO: 2430) HPV 16 E7 11-20

A*02:01 KLPQLCTEL (SEQ ID NO: 2431) HPV 16 E6 18-26

A*02:01 YMLDLQPET (SEQ ID NO: 2432) HPV 16 E7 11-19

A*02:01 MLDLQPETT (SEQ ID NO: 2433) HPV 16 E7 12-20

A*02:01 VLMIKALEL (SEQ ID NO: 2434) Non muscle

Myosin-9 741-749

A*02:01 QLFNHTMFI (SEQ ID NO: 2435) Non-muscle

Myosin 478-486

A*02:01 QMARLAWEA (SEQ ID NO: 2436) 1116-1124

A*02:01 LLFGYPVYV (SEQ ID NO: 2437) Human T-cell

lymphotropic virus-1 (HTLV-1) tax 11-19

A*02:01 AVLDGLLSL (SEQ ID NO: 2438) HTLV bZIP

factor 42-50

A*02:01 GLLSLEEEL (SEQ ID NO: 2439) bZIP factor

26-34

A*02:01 GILGFVFTL (SEQ ID NO: 2440) Influenza A

MP 58-66

A*02:01 ILGFVFTLTV (SEQ ID NO: 2441) Influenza A

MP 59-68

A*02:01 KLGEFYNQMM (SEQ ID NO: 2442) Flu BNP

85-94 (Influenza B)

A*02:01 SITEVECFL (SEQ ID NO: 2443) VP1 36-44

A*02:01 ILMWEAVTL (SEQ ID NO: 2444) VP1 100-108

A*02:01 ALPHIIDEV (SEQ ID NO: 2445) LCMV envelope

gp 10-18

A*02:01 YLVSIFLHL (SEQ ID NO: 2446) LCMV envelope

gp 447-455

A*02:01 SLNQTVHSL (SEQ ID NO: 2447) NP 69-77

A*02:01 YLNKIQNSL (SEQ ID NO: 2448) Plasmodium

falciparum CSP 334-342

A*02:01 FIDSYICQV (SEQ ID NO: 2449) miHAg H-Y

(human SMCY) 311-319

A*02:01 YIGEVLVSV (SEQ ID NO: 2450) HA-2

A*02:01 VLHDDLLEA (SEQ ID NO: 2451) Minor

Histocompatibility Antigen HA-1 137-145

A*02:01 RTLDKVLEV (SEQ ID NO: 2452) miHAg HA-8

A*02:01 NEGATIVE (SEQ ID NO: 2453) Negative Control

A*02:01 TMFPHIIVDV (SEQ ID NO: 2454) Norovirus VP1

139-148

A*02:01 LLDVPTAAV (SEQ ID NO: 2455) Interferon

gamma inducible protein (GILT) 30 27-35

A*02:01 RILGAVAKV (SEQ ID NO: 2456) Vinculin

822-830

A*02:01 LMWYELSKI (SEQ ID NO: 2457) KSHVF-8

gB.492-500

A*02:01 ILEDIVLTL (SEQ ID NO: 2458) Streptococcus

pyogenes Cas9 615-623

A*02:01 KMLKEMGEV (SEQ ID NO: 2459) RSV NP 137-145

A*02:01 KLIANNTRV (SEQ ID NO: 2460) Mycobacterium

bovis antigen 85-A 200-208

A*02:01 GLPVEYLQV (SEQ ID NO: 2461) Mycobacterium

bovis antigen 85-A 6-14

A*02:01 GILTVSVAV (SEQ ID NO: 2462) 16 kDa

A*02:01 AMASTEGNV (SEQ ID NO: 2463) ESAT-6

A*02:01 VLTDGNPPEV (SEQ ID NO: 2464) 19 kDa

A*02:01 KVDDTFYYV (SEQ ID NO: 2465) Vaccinia virus

Host range protein 2 74-82

A*02:01 ILDDNLYKV (SEQ ID NO: 2466) Vaccinia virus

Copenhagen Protein G5 18-26

A*02:01 ALWALPHAA (SEQ ID NO: 2467) IE62 593-601

A*02:01 RLDDDGNFQL (SEQ ID NO: 2468) West Nile

Virus NY-99 polyprotein precursor (1452-1461)

A*02:01 ATWAENIQV (SEQ ID NO: 2469) West Nile

virus NY-99 polyprotein precursor 3390-3398

A*02:01 YTMDGEYRL (SEQ ID NO: 2470) West Nile

virus NY-99 polyprotein precursor 2023-2031

A*02:01 SVGGVFTSV (SEQ ID NO: 2471) WNV envelope

gp 430-438

A*02:01 SLFGQRIEV (SEQ ID NO: 2472) WNV

nonstructural protein 4B 15-23

A*02:01 LLWNGPMAV (SEQ ID NO: 2473) NS4B 214-222

A*03:01 KQSSKALQR (SEQ ID NO: 2474) bcr-abl 210 kD fusion

protein 21-29

A*03:01 ALLAVGATK (SEQ ID NO: 2475) gp100 (pmel17) 17-25

A*03:01 ATGFKQSSK (SEQ ID NO: 2476) bcr-abl 210 kD fusion

protein 259-269

A*03:01 RISTFKNWPK (SEQ ID NO: 2477) Survivin-3A 18-27 (27K)

A*03:01 RLGLQVRKNK (SEQ ID NO: 2478) RhoC 176-185 (177L)

A*03:01 RLLFFAPTR (SEQ ID NO: 2479) Mcl-1 95-103

A*03:01 QVLKKIAQK (SEQ ID NO: 2480) HMOX1 145-153

A*03:01 RIAAWMATY (SEQ ID NO: 2481) 165-173

A*03:01 KLGGALQAK (SEQ ID NO: 2482) HCMV IE1 184-192

A*03:01 KTFPPTEPK (SEQ ID NO: 2483) SARS-CoV-2 Nucleocapsid

protein 362-370 (confirmed epitope)

A*03:01 ELERAADVK (SEQ ID NO: 2484) Dengue NS2b 52-60

A*03:01 RVSTVQQLTK (SEQ ID NO: 2485) Dengue C 22-31

A*03:01 RIEPSWADVK (SEQ ID NO: 2486) Dengue NS3 64-74

A*03:01 RVIDPRRCMK (SEQ ID NO: 2487) Dengue NS3 422-431

A*03:01 KITAEWLWK (SEQ ID NO: 2488) Dengue NS5 375-383

A*03:01 RLRAEAQVK (SEQ ID NO: 2489) EBV EBNA 3A 603-611

A*03:01 RVRAYTYSK (SEQ ID NO: 2490) EBV BRLF1

A*03:01 RVCEKMALY (SEQ ID NO: 2491) HCV NS5B 2588-2596

A*03:01 QVPLRPMTYK (SEQ ID NO: 2492) HIV-1 nef 73-82

A*03:01 RLRPGGKKK (SEQ ID NO: 2493) HIV-1 gag p17 19-27

A*03:01 AIFQSSMTK (SEQ ID NO: 2494) HIV pol 325-333

A*03:01 KLCLRFLSK (SEQ ID NO: 2495) HPV 33 E6 64-72

A*03:01 ILRGSVAHK (SEQ ID NO: 2496) Influenza A (PR8) NP

265-274

A*11:01 KTFPPTEPK (SEQ ID NO: 2483) SARS-CoV-2 Nucleocapsid

protein 362-370 (confirmed epitope)

A*11:01 GTSGSPIINR (SEQ ID NO: 2497) Dengue NS3 serotype 3&4

133-142

A*11:01 GTSGSPIIDK (SEQ ID NO: 2498) Dengue NS3 133-142

A*11:01 GTSGSPIVNR (SEQ ID NO: 2499) NS3 serotype 1 133-142

A*11:01 GTSGSPIVDR (SEQ ID NO: 2500) Dengue NS3 serotype 2

133-142

A*11:01 GTSGSPIADK (SEQ ID NO: 2501) Dengue NS3 133-142

A*11:01 RVSTVQQLTK (SEQ ID NO: 2485) Dengue C 22-31

A*11:01 RIEPSWADVK (SEQ ID NO: 2486) Dengue NS3 64-74

A*11:01 RVIDPRRCMK (SEQ ID NO: 2487) Dengue NS3 422-431

A*11:01 KITAEWLWK (SEQ ID NO: 2488) Dengue NS5 375-383

A*11:01 IVTDFSVIK (SEQ ID NO: 2502) EBV EBNA-4 416-424

A*11:01 SSCSSCPLSK (SEQ ID NO: 2503) EBV LMP-2 340-349

A*11:01 ATIGTAMYK (SEQ ID NO: 2504) EBV BRLF1 134-142

A*11:01 AVFDRKSDAK (SEQ ID NO: 2505) EBNA3B 399-408

A*11:01 YVNVNMGLK (SEQ ID NO: 2506) HBV core antigen 88-96

A*11:01 YVNTNMGLK (SEQ ID NO: 2507) HBV core 88-96

A*11:01 STLPETTVVRR (SEQ ID NO: 2508) HBV core 141-151

A*11:01 AVDLSHFLK (SEQ ID NO: 2509) HIV nef 84-92

A*11:01 ACQGVGGPGHK (SEQ ID NO: 2510) HIV gag p24

A*11:01 NTLEQTVKK (SEQ ID NO: 2511) HPV 33 E6 86-94

A*11:01 SIIPSGPLK (SEQ ID NO: 2512) Influenza A MP 13-21

A*11:01 RMVLASTTAK (SEQ ID NO: 2513) Influenza A MP1 178-187

A*11:01 KSMREEYRK (SEQ ID NO: 2514) Influenza A MP2 70-78

A*24:02 TYFSLNNKF (SEQ ID NO: 2515) Adenovirus 5 Hexon 37-45

A*24:02 TYACFVSNL (SEQ ID NO: 2516) Carcinogenic Embryonic

Antigen (CEA) 652-660

A*24:02 AFLPWHRLF (SEQ ID NO: 2517) Tyrosinase 188-196

A*24:02 IMPKAGLLI (SEQ ID NO: 2518) MAGE-A3

A*24:02 VYFFLPDHL (SEQ ID NO: 2519) gp100-intron 4 (170-178)

A*24:02 EYLQLVFGI (SEQ ID NO: 2520) MAGEA2 156-164

A*24:02 TYLPTNASL (SEQ ID NO: 2521) HER-2/neu 63-71

A*24:02 VYGFVRACL (SEQ ID NO: 2522) Telomerase reverse

transcriptase (hTRT) 461-469

A*24:02 TFPDLESEF (SEQ ID NO: 2523) MAGEA3 97-105

A*24:02 DYLQYVLQI (SEQ ID NO: 2524) MiHA ACC1 15-23

A*24:02 RYCNLEGPPI (SEQ ID NO: 2525) Lymphocyte antigen 6

complex locus K (LY6K) 177-186

A*24:02 AYACNTSTL (SEQ ID NO: 2526) Survivin 80-88

A*24:02 CYASGWGSI (SEQ ID NO: 2527) Prostate Specific

Antigen-1 153-161

A*24:02 DYLNEWGSRF (SEQ ID NO: 2528) CDH3 807-816

A*24:02 EYCPGGNLF (SEQ ID NO: 2529) MELK 87-95 (93N)

A*24:02 EYYELFVNI (SEQ ID NO: 2530) DEP DC1 294-302

A*24:02 GYCTQIGIF (SEQ ID NO: 2531) HENMT1 221-229

A*24:02 IYTWIEDHF (SEQ ID NO: 2532) FOXM1 262-270

A*24:02 NYQPVWLCL (SEQ ID NO: 2533) RNF43 721-729 (722Y)

A*24:02 RYNAQCQETI (SEQ ID NO: 2534) Midkine 110-119

A*24:02 EYRALQLHL (SEQ ID NO: 2535) CA9 219-227

A*24:02 SYRNEIAYL (SEQ ID NO: 2536) TTK protein kinase 551-559

A*24:02 VYLRVRPLL (SEQ ID NO: 2537) KIF20A 67-75

A*24:02 VYYNWQYLL (SEQ ID NO: 2538) IL13r 146-154

A*24:02 VYALPLKML (SEQ ID NO: 2539) HCMV pp65 113-121

A*24:02 QYDPVAALF (SEQ ID NO: 2540) HCMV pp65 341-349

A*24:02 AYAQKIFKI (SEQ ID NO: 2541) CMV IE-1 248-256

A*24:02 QYSDRRWCF (SEQ ID NO: 2542) Dengue NS3 557-565

(Singapore/S275/1990)

A*24:02 TYGPVFMCL (SEQ ID NO: 2543) EBV LMP-2 419-427

A*24:02 PYLFWLAAI (SEQ ID NO: 2544) EBV LMP2 131-139

A*24:02 TYGPVFMSL (SEQ ID NO: 2545) EBV LMP2 419-427

A*24:02 EYLVSFGVW (SEQ ID NO: 2546) HBV core 117-125

A*24:02 KYTSFPWLL (SEQ ID NO: 2547) HBV polymerase 756-764

A*24:02 FFPSIRDLL (SEQ ID NO: 2548) HBV core protein 23-31

A*24:02 AYSQQTRGL (SEQ ID NO: 2549) HCV NS3 1031-1039

A*24:02 RYPLTFGWCY (SEQ ID NO: 2550) HIV-1 Nef 134-143

A*24:02 RYLKDQQLL (SEQ ID NO: 2551) HIV-1 gag gp41 67-75

A*24:02 RYLRDQQLL (SEQ ID NO: 2552) HIV env

A*24:02 RYPLTFGWCF (SEQ ID NO: 2553) HIV nef 143-152

A*24:02 RYPLTFGW (SEQ ID NO: 2554) HIV nef

A*24:02 VYDFAFRDL (SEQ ID NO: 2555) HPV16 E6

A*24:02 SFHSLHLLF (SEQ ID NO: 2556) HTLV Tax 301-309

A*29:02 KEKYIDQEEL (SEQ ID NO: 2557) HSP90 alpha 280-288

(Pathologic Conditions)

A*29:02 LYNTVATLY (SEQ ID NO: 2558) HIV gag 79-86

A*29:02 SFDPIPIHY (SEQ ID NO: 2559) HIV env 216-224

A*29:02 SFNCRGEFFY (SEQ ID NO: 2560) HIV env 382-391

A*68:01 TVSGNILTIR (SEQ ID NO: 2561) NY-ESO-1 127-136

B*07:02 VPQYGYLTL (SEQ ID NO: 2562) AAV2 372-380

B*07:02 KPYSGTAYNSL (SEQ ID NO: 2563) Adenovirus Hexon 114-124

B*07:02 KPYSGTAYNAL (SEQ ID NO: 2564) Adenovirus Hexon 114-124

B*07:02 LPLMRKAYL (SEQ ID NO: 2565) LT antigen 27-35

B*07:02 LPWHRLFLL (SEQ ID NO: 2566) Tyrosinase 208-216

B*07:02 EPR(PHOSPHO-S)PSHSM (SEQ ID NO: 2567) Insulin receptor

substrate 2

B*07:02 TPNQRQNVC (SEQ ID NO: 2568) P2X5

B*07:02 APRGVRMAV (SEQ ID NO: 2569) LAGE-1 46-54

B*07:02 LPVSPRLQL (SEQ ID NO: 2570) CEACAM 185-193

B*07:02 TPRVTGGGAM (SEQ ID NO: 2571) HCMV pp65 417-426

B*07:02 RPHERNGFTVL (SEQ ID NO: 2572) HCMV pp65 265-275

B*07:02 SPRWYFYYL (SEQ ID NO: 2573) SARS-CoV-2 Nucleocapsid

protein 105-113 (confirmed epitope)

B*07:02 APTRVVAAEM (SEQ ID NO: 2574) Dengue NS3 serotype 2

222-231

B*07:02 RPPIFIRRL (SEQ ID NO: 2575) EBV EBNA-3A 247-255

B*07:02 RPQGGSRPEFVKL (SEQ ID NO: 2576) EBV BMRF1 116-128

B*07:02 QPRAPIRPI (SEQ ID NO: 2577) EBV EBNA-3C 881-889

B*07:02 LPSDFFPSV (SEQ ID NO: 2578) HBV core 19-27

B*07:02 GPRLGVRAT (SEQ ID NO: 2579) HCV core 41-49

B*07:02 DPRRRSRNL (SEQ ID NO: 2580) HCV core 111-119

B*07:02 IPRRIRQGL (SEQ ID NO: 2581) HIV-1 env gp120 848-856

B*07:02 TPGPGVRYPL (SEQ ID NO: 2582) HIV-1 nef 128-137

B*07:02 GPGHKARVL (SEQ ID NO: 2583) HIV gag p24 223-231

B*07:02 KPTLKEYVL (SEQ ID NO: 2584) HPV 33 E7 5-13

B*07:02 LPVSCPEDL (SEQ ID NO: 2585) bZIP factor 10-18

B*07:02 QPEWFRNVL (SEQ ID NO: 2586) Influenza A PB1 329-337

B*07:02 SPIVPSFDM (SEQ ID NO: 2587) Influenza A NP 473-481

B*07:02 SPSVDKARAEL (SEQ ID NO: 2588) MiHAg SMCY 1041-1051

B*08:01 GFKQSSKAL (SEQ ID NO: 2589) bcr-abl 210 kD fusion

protein 19-27

B*08:01 ELRRKMMYM (SEQ ID NO: 2590) IE1 199-207

B*08:01 ELKRKMIYM (SEQ ID NO: 2591) CMV IE-1 199-207 (R/L

Position 3, M/I Position 7)

B*08:01 RIKQKGIL (SEQ ID NO: 2592) Dengue NS3 25-32

B*08:01 LEKTKKDL (SEQ ID NO: 2593) Dengue NS4a 6-13

B*08:01 FLRGRAYGL (SEQ ID NO: 2594) EBV EBNA-3A 193-201

B*08:01 RAKFKQLL (SEQ ID NO: 2595) EBV BZLF-1 190-197

B*08:01 QAKWRLQTL (SEQ ID NO: 2596) EBV EBNA3A 158-166

B*08:01 GLKILQLL (SEQ ID NO: 2597) HBV external core Ag

B*08:01 HSKKKCDEL (SEQ ID NO: 2598) HCV NS3 1395-1403

B*08:01 FLKEKGGL (SEQ ID NO: 2599) HIV-1 nef 90-97

B*08:01 GEIYKRWII (SEQ ID NO: 2600) HIV-1 gag p24 261-269

B*08:01 EIYKRWII (SEQ ID NO: 2601) HIV p24 gag 128-135

B*08:01 YLKDQQLL (SEQ ID NO: 2602) Env 586-593

B*15:01 CLIPTAMAF (SEQ ID NO: 2603) Dengue C 107-115

B*15:01 RLRPGGKKKY (SEQ ID NO: 2604) HIV-1 p17 20-29

B*27:05 GRFGLATEK (SEQ ID NO: 2605) BRAF 594-601 (600E)

B*27:05 GRFGLATVK (SEQ ID NO: 2606) BRAF 594-601 (600V)

B*27:05 RMFPNAPYL (SEQ ID NO: 2362) WT-1 126-134 (Wilms tumor)

B*27:05 ARKLLLDNL (SEQ ID NO: 2607) PqqC-like protein 70-78

B*27:05 NRAKQVIKL (SEQ ID NO: 2608) Probable ATP-dependent Clp

protease ATP-binding subunit 7-15

B*27:05 QRNAPRITF (SEQ ID NO: 2609) SARS-CoV-2 Nucleocapsid

protein 9-17 (confirmed epitope)

B*27:05 ARMILMTHF (SEQ ID NO: 2610) HCV NS5B 2841-2849

B*27:05 KRWIILGLNK (SEQ ID NO: 2611) HIV-1 gag p24 265-274

B*27:05 KRWIIMGLNK (SEQ ID NO: 2612) HIV-1 Gag p24 263-272

B*27:05 GRAFVTIGK (SEQ ID NO: 2613) HIV-1 gp100 103-111

B*27:05 SRYWAIRTR (SEQ ID NO: 2614) Influenza A NP 383-391

B*35:01 IPYLDGTFY (SEQ ID NO: 2615) Adenovirus Hexon

B*35:01 MPFATPMEA (SEQ ID NO: 2616) NY-ESO-1 94-102

B*35:01 IPSINVHHY (SEQ ID NO: 2617) HCMV pp65 123-131

B*35:01 TPEGIIPTL (SEQ ID NO: 2618) Dengue NS3 500-508

B*35:01 YPLHEQHGM (SEQ ID NO: 2619) EBV EBNA-3A 458-466

B*35:01 EPLPQGQLTAY (SEQ ID NO: 2620) EBV BZLF-1 54-64

B*35:01 HPVAEADYFEY (SEQ ID NO: 2621) EBV EBNA-1 407-417(4A)

B*35:01 HPNIEEVAL (SEQ ID NO: 2622) HCV NS3 1359-1367

B*35:01 CPNSSIVY (SEQ ID NO: 2623) HCV E1 207-214

B*35:01 NPDIVIYQY (SEQ ID NO: 2624) HIV-1 RT 330-338

B*35:01 NPDIVIYQY (SEQ ID NO: 2625) HIV-1 HIV-1 RT 328-336

B*35:01 VPLDEDFRKY (SEQ ID NO: 2626) HIV-1 HIV-1 RT 273-282

B*40:01 MEVTPSGTWL (SEQ ID NO: 2627) SARS-CoV-2 Nucleocapsid

protein 322-330 (confirmed epitope)

B*40:01 GEARKTFVEL (SEQ ID NO: 2628) Dengue NS3 528-537

B*40:01 IEDPPFNSL (SEQ ID NO: 2629) EBV LMP2 200-208

B*40:01 REISVPAEIL (SEQ ID NO: 2630) HCV NS5a 2266-2275

B*40:01 KEKGGLEGL (SEQ ID NO: 2631) HIV-1 Nef 92-100

C*06:02 TRATKMQVI (SEQ ID NO: 2632) pp65 211-219

C*06:02 QIKVRVDM (SEQ ID NO: 2633) IE1 88-95

C*06:02 TRRFLPQIL (SEQ ID NO: 2634) NS3 205-213

C*07:02 CRVLCCYVL (SEQ ID NO: 2635) IE1 309-317

E*01:01 VMAPRTLIL (SEQ ID NO: 2636) HLA-C leader sequence

peptide

E*01:01 VMAPRTLVL (SEQ ID NO: 2637) HLA-A leader sequence

peptide

A*01:01 SADNNNSEY (SEQ ID NO: 2082) AAV VP1 492-500

A*01:01 TDLGQNLLY (SEQ ID NO: 2083) Adenovirus 5 Hexon 886-894

A*01:01 EADPTGHSY (SEQ ID NO: 2084) MAGE-A1 161-169

A*01:01 EVDPIGHLY (SEQ ID NO: 2085) MAGE-A3 168-176

A*01:01 KSDICTDEY (SEQ ID NO: 2086) Tyrosinase 243-251 (244S)

A*01:01 KCDICTDEY (SEQ ID NO: 2087) Tyrosinase 243-251

A*01:01 QSLEIISRY (SEQ ID NO: 2088) Mcl-1 177-185

A*01:01 YVDFREYEYY (SEQ ID NO: 2089) FLT3 ITD

A*01:01 TLDTLTAFY (SEQ ID NO: 2090) Mesothelin 429-437

A*01:01 LTDDRLFTCY (SEQ ID NO: 2091) PLEKHM2

A*01:01 DSDPDSFQDY (SEQ ID NO: 2092) Tyr A1a 454-463

A*01:01 EADPIGHLY (SEQ ID NO: 2093) MAGEA3

A*01:01 EVDPASNTY (SEQ ID NO: 2094) MAGE-A4 169-177

A*01:01 HSTNGVTRIY (SEQ ID NO: 2095) PSMA

A*01:01 ILDTAGREEY (SEQ ID NO: 2096) N-ras 55-64

A*01:01 LVDVMPWLQY (SEQ ID NO: 2097) Cytochrome P450 240-249

A*01:01 RSDSGQQARY (SEQ ID NO: 2098) AIM-2

A*01:01 VTEPGTAQY (SEQ ID NO: 2099) Minor antigen HA-3T (Lbc

oncogene 451-459)

A*01:01 VYDFFVWLHY (SEQ ID NO: 2100) TRP-2 181-190

A*01:01 YSEHPTFTSQY (SEQ ID NO: 2101) HCMV pp65 363-373

A*01:01 VTEHDTLLY (SEQ ID NO: 2102) HCMV pp50 245-253

A*01:01 FTSDYYQLY (SEQ ID NO: 2103) SARS-CoV-2 ORF3a 207-215

(confirmed epitope)

A*01:01 TTDPSFLGRY (SEQ ID NO: 2638) SARS-CoV-2 Replicase

polyprotein 1ab 1637-1646 (confirmed epitope)

A*01:01 PTDNYITTY (SEQ ID NO: 2639) SARS-CoV-2 Replicase

polyprotein 1ab 1621-1629 (confirmed epitope)

A*01:01 LLDTASALY (SEQ ID NO: 2106) HBV core 30-38

A*01:01 ATDALMTGY (SEQ ID NO: 2107) HCV NS3 1435-1443

A*01:01 ATDALMTGF (SEQ ID NO: 2108) HCV NS3 1436-1444

A*01:01 CTELKLSDY (SEQ ID NO: 2109) Influenza A (PR8) NP 44-52

A*01:01 VSDGGPNLY (SEQ ID NO: 2110) Influenza A PB1 591-599

A*01:01 IVDCLTEMY (SEQ ID NO: 2111) DRRFY (1521-1529))

A*0201 ALCNTDSPL (SEQ ID NO: 2112) iLR1

A*0201 ALKDVEERV (SEQ ID NO: 2113) MAGE-C2 336-344

A*0201 LLAARAIVAI (SEQ ID NO: 2114) iLR1 59-68

A*0201 RLWQELSDI (SEQ ID NO: 2115) circadian clock protein

PASD1 691-700

A*0201 LLFGLALIEV (SEQ ID NO: 2116) MAGE-C2 191-200

A*0201 FLDPRPLTV (SEQ ID NO: 2117) CYP190

A*0201 STLCQVEPV (SEQ ID NO: 2118) MPP11

A*0201 VLQMKEEDV (SEQ ID NO: 2119) ILR1

A*0201 AIQDLCLAV (SEQ ID NO: 2120) NPM1

A*0201 QLLIKAVNL (SEQ ID NO: 2121) MPP11

A*0201 AIQDLCVAV (SEQ ID NO: 2122) NPM1

A*0201 ALTPVVVTL (SEQ ID NO: 2123) cyclin-dependent kinase 4

170-178

A*02:01 KLQVFLIVL (SEQ ID NO: 2640) T1D Diabetes human prepro

islet amyloid polypeptide pplAPP 5-13

A*02:01 VMNILLQYV (SEQ ID NO: 2125) GAD65 114-123

A*02:01 SLSRFSWGA (SEQ ID NO: 2126) Myelin basic protein

110-118

A*02:01 HLVEALYLV (SEQ ID NO: 2127) Insulin B chain 10-18

A*02:01 LNIDLLWSV (SEQ ID NO: 2128) T1D Diabetes IGRP 228-236

A*02:01 VLFGLGFAI (SEQ ID NO: 2129) T1D Diabetes IGRP 265-273

A*02:01 ALWGPDPAAA (SEQ ID NO: 2130) Proinsulin precursor 15-24

A*02:01 MVWESGCTV (SEQ ID NO: 2131) IA-2 797-805

A*02:01 YTCPLCRAPV (SEQ ID NO: 2132) SSA SS-56 55-64

A*02:01 VIVMLTPLV (SEQ ID NO: 2133) IA-2 805-813

A*02:01 AITEVECFL (SEQ ID NO: 2134) VP1 44-52

A*02:01 FLHCIVFNV (SEQ ID NO: 2135) large T antigen 410-418

A*02:01 LLMWEAVTV (SEQ ID NO: 2136) VP1 108-116

A*02:01 CLLPKMDSV (SEQ ID NO: 2137) large T antigen 398-406

A*02:01 FLWGPRALV (SEQ ID NO: 2138) MAGEA3 271-279

A*02:01 IMDQVPFSV (SEQ ID NO: 2139) gp100 (pmel17) 209-217

A*02:01 YLEPGPVTV (SEQ ID NO: 2140) gp100 (pmel) 280-288 (288V)

A*02:01 YLSGADLNL (SEQ ID NO: 2141) Carcinoembryonic antigen

(CEA)-derived peptide CAP1-6D

A*02:01 SLLMWITQC (SEQ ID NO: 2142) NY-ESO-1 157-165 (9C)

A*02:01 KTWGQYWQV (SEQ ID NO: 2143) gp100 (pmel17) 154-162

A*02:01 YLEPGPVTA (SEQ ID NO: 2144) gp100

A*02:01 YMDGTMSQV (SEQ ID NO: 2145) Tyrosinase 369-377 (371D)

A*02:01 YLSGANLNL (SEQ ID NO: 2146) Carcinogenic Embryonic

Antigen (CEA) 571-579

A*02:01 ELAGIGILTV (SEQ ID NO: 2147) MelanA/MART 26-35

A*02:01 ILAKFLHWL (SEQ ID NO: 2148) Telomerase 540-548

A*02:01 ALQPGTALL (SEQ ID NO: 2149) Prostate Stem Cell Antigen

(PSCA) 14-22

A*02:01 VISNDVCAQV (SEQ ID NO: 2150)Prostate Specific

Antigen-1 (PSA-1) 154-163

A*02:01 RLVDDFLLV (SEQ ID NO: 2151) Telomerase Reverse

Transcriptase 865-873

A*02:01 GVLVGVALI (SEQ ID NO: 2152) Carcinogenic Embryonic

Antigen (CEA) 694-702

A*02:01 VLYRYGSFSV (SEQ ID NO: 2153) gp100 (pmel17) 476-485

A*02:01 PLFQVPEPV (SEQ ID NO: 2154) Alpha-fetoprotein isoform

1 137-145

A*02:01 FMNKFIYEI (SEQ ID NO: 2155) Human alfa fetoprotein

158-166

A*02:01 GLSPNLNRFL (SEQ ID NO: 2156) Alpha-fetoprotein isoform

2 167-176

A*02:01 KVLEYVIKV (SEQ ID NO: 2157) MAGEA1 278-286

A*02:01 LLGRNSFEV (SEQ ID NO: 2158) p53 264-272

A*02:01 LLLLTVLTV (SEQ ID NO: 2159) MUC-1 12-20

A*02:01 ILHNGAYSL (SEQ ID NO: 2160) HER-2/neu 435-443

A*02:01 RLLQETELV (SEQ ID NO: 2161) HER-2/neu 689-697

A*02:01 KIFGSLAFL (SEQ ID NO: 2162) HER-2/neu 369-377

A*02:01 LLLLDVAPL (SEQ ID NO: 2163) HSP1A 459-467

A*02:01 LLDVAPLSL (SEQ ID NO: 2164) HSP1A 461-469

A*02:01 HLYQGCQVV (SEQ ID NO: 2165) Receptor tyrosine-protein

kinase erbB-2 48-56

A*02:01 HLSTAFARV (SEQ ID NO: 2166) G250 (renal cell

carcinoma) 217-225

A*02:01 VLQELNVTV (SEQ ID NO: 2167) Leukocyte Proteinase-3

(Wegener's autoantigen) 169-177

A*02:01 KVAELVHFL (SEQ ID NO: 2168) MAGEA3 112-120

A*02:01 VLAGVGFFI (SEQ ID NO: 2169) EPHA2 550-558

A*02:01 FLYTLLREV (SEQ ID NO: 2170) STEAP 86-94

A*02:01 ILLWQPIPV (SEQ ID NO: 2171) Prostatic Acid

Phosphatase-3 (PAP-3) 135-143

A*02:01 RLQEERTCKV (SEQ ID NO: 2172) BIR

A*02:01 QLCPICRAPV (SEQ ID NO: 2173) Livin/ML-IAP280 175-184

A*02:01 VLGEAWRDQV (SEQ ID NO: 2174) TRAP 45-54

A*02:01 LLLTVLTVV (SEQ ID NO: 2175) Tumor Mucin Antigen 13-21

A*02:01 GLYDGMEHL (SEQ ID NO: 2176) MAGEA-10 254-262

A*02:01 SLLMWITQV (SEQ ID NO: 2177) NY-ESO-1 157-165

A*02:01 LMLGEFLKL (SEQ ID NO: 2178) Survivin 96-104

A*02:01 YLFFYRKSV (SEQ ID NO: 2179) mTERT 572-580

A*02:01 ELTLGEFLKL (SEQ ID NO: 2180) survivin 95-104

A*02:01 FLTPKKLQCV (SEQ ID NO: 2181) Prostate Specific

Antigen-1 (PSA-1) 141-150

A*02:01 KLQCVDLHV (SEQ ID NO: 2182) Prostate Specific Antigen

146-154

A*02:01 TLAPATEPA (SEQ ID NO: 2183) Mucin 79-87

A*02:01 YLQVNSLQTV (SEQ ID NO: 2184) Telomerase Reverse

Transcriptase (hTRT) 988-997

A*02:01 SLGEQQYSV (SEQ ID NO: 2185) WT1 187-195

A*02:01 SLEENIVIL (SEQ ID NO: 2186) RHAMM 275-283

A*02:01 YMNGTMSQV (SEQ ID NO: 2187) Tyrosinase 368-376

A*02:01 ILSLELMKL (SEQ ID NO: 2188) Receptor for hyaluronic

acid-mediated motility (RHAMM) 165-173

A*02:01 PLFDFSWLSL (SEQ ID NO: 2189) Bcl-2 208-217

A*02:01 LLGATCMFV (SEQ ID NO: 2190) CyclinD 101-109

A*02:01 ALYVDSLFFL (SEQ ID NO: 2191) PRAME PRA 300-309

A*02:01 GLMEEMSAL (SEQ ID NO: 2192) Human Mena protein

(overexpressed in breast cancer)

A*02:01 TMNGSKSPV (SEQ ID NO: 2193) hMena 502-510

A*02:01 GVYDGREHTV (SEQ ID NO: 2194) MAGE-A4 230-239

A*02:01 YLNDHLEPWI (SEQ ID NO: 2195) Bcl-X 173-182

A*02:01 ALDVYNGLL (SEQ ID NO: 2196) Prostatic acid phosphatase

precursor (PAP) 299-307

A*02:01 ALFDIESKV (SEQ ID NO: 2197) PSM P2 (prostate)

A*02:01 SLAMLDLLHV (SEQ ID NO: 2198) Mutant anaplastic

lymphoma kinase 1220-1229

A*02:01 YLNTVQPTCV (SEQ ID NO: 2199) EGF-R 1138-1147

A*02:01 KLFGTSGQKT (SEQ ID NO: 2200) EGF-R-479 350-359

A*02:01 RMPEAAPPV (SEQ ID NO: 2201) p53 65-73

A*02:01 PLTSIISAV (SEQ ID NO: 2202) Receptor tyrosine-protein

kinase erbB-2 728-736

A*02:01 VLAGGFFLL (SEQ ID NO: 2203) PSMA 27-38

A*02:01 LLHETDSAV (SEQ ID NO: 2204) PSMA/PSM-P1 4-12

A*02:01 VMAGVGSPYV (SEQ ID NO: 2205) Receptor tyrosine-protein

kinase erbB-2 819-828

A*02:01 VLPLTVAEV (SEQ ID NO: 2206) Mesothelin 530-538

A*02:01 SLLFLLFSL (SEQ ID NO: 2207) Mesothelin 20-28

A*02:01 QLFEELQEL (SEQ ID NO: 2208) Heme oxygenase-1 212-220

A*02:01 VLDGLDVLL (SEQ ID NO: 2209) PRAME 100-108

A*02:01 RLASFYDWPL (SEQ ID NO: 2210) BIR7 90-99

A*02:01 LIAHNQVRQV (SEQ ID NO: 2211) HER-2/neu (85-94)

A*02:01 ILHDGAYSL (SEQ ID NO: 2212) HER-2 434-443

A*02:01 FVGEFFTDV (SEQ ID NO: 2213) GPC3 144-152

(overexpressed in hepatocellular carcinoma)

A*02:01 LLLIWFRPV (SEQ ID NO: 2214) BKV Ltag 579-587

A*02:01 KLQDASAEV (SEQ ID NO: 2215) HM1.24-aa 126-134

A*02:01 SLYSFPEPEA (SEQ ID NO: 2216) PRAME

A*02:01 SLLQHLIGL (SEQ ID NO: 2217) PRAME 425-433

A*02:01 VIFDFLHCI (SEQ ID NO: 2218) BKV Ltag 406-414

A*02:01 VLDFAPPGA (SEQ ID NO: 2219) WT1

A*02:01 TLPGYPPHV (SEQ ID NO: 2220) PAX-5 311-319

A*02:01 YMEHNNVYTV (SEQ ID NO: 2221) Fibromodulin 250-259

A*02:01 YLQHNEIQEV (SEQ ID NO: 2222) Fibromodulin 206-215

A*02:01 SLVDVMPWL (SEQ ID NO: 2223) Cytochrome p450 1B1 239-248

A*02:01 RLMNDMTAV (SEQ ID NO: 2224) HSP105 128-136

A*02:01 RLARLALVL (SEQ ID NO: 2225) Trophoblast glycoprotein

17-25

A*02:01 FLTGNQLAV (SEQ ID NO: 2226) 5T4 97-105

A*02:01 LLLAGLFSL (SEQ ID NO: 2227) Fibromodulin 7-15

A*02:01 FLGYLILGV (SEQ ID NO: 2228) Prostatic Acid Phosphatase-

3 (PAP-3)

A*02:01 SLFLGILSV (SEQ ID NO: 2229) CD20 188-196 (B cell

malignancies)

A*02:01 AVLPLLELV (SEQ ID NO: 2230) MCL-1 139-147

A*02:01 SLSEKTVLL (SEQ ID NO: 2231) CD59 glycoprotein precursor

106-114

A*02:01 YMCSFLFNL (SEQ ID NO: 2232) Ewing Tumor EZH2 666-674

A*02:01 YLISGDSPV (SEQ ID NO: 2233) CD33 65-73 (1Y2L)

A*02:01 KASEKIFYV (SEQ ID NO: 2234) SSX2 41-49

A*02:01 FLAKLNNTV (SEQ ID NO: 2235) HCA587 317-325

A*02:01 GLAPPQHLIRV (SEQ ID NO: 2236) p53 187-197

A*02:01 VIMPCSWWV (SEQ ID NO: 2237) Chondromodulin-I 319-327

A*02:01 KVVEFLAML (SEQ ID NO: 2238) MAGE-C1 1083-1091

A*02:01 LTLGEFLKL (SEQ ID NO: 2239) Survivin-3A 96-104

A*02:01 ALPFGFILV (SEQ ID NO: 2240) IL13R 345-353

A*02:01 TLADFDPRV (SEQ ID NO: 2241) EphA2

A*02:01 ALMEQQHYV (SEQ ID NO: 2242) ITGB8 662-670

A*02:01 CLTSTVQLV (SEQ ID NO: 2243) HER-2/neu 789-797

A*02:01 GLLGASVLGL (SEQ ID NO: 2244) Telomerase Reverse

Transcriptase (hTRT) 674-683

A*02:01 QLLDGFMITL (SEQ ID NO: 2245) PASD1 39-48

A*02:01 YLVGNVCIL (SEQ ID NO: 2246) PASD1 168-176

A*02:01 ALLTSRLRFI (SEQ ID NO: 2247) Telomerase Reverse

Transcriptase (hTRT) 615-624

A*02:01 RLSSCVPVA (SEQ ID NO: 2248) TGF beta receptor type-2

131-139

A*02:01 FLYDDNQRV (SEQ ID NO: 2249) Topoisomerase II-alpha-b

828-836

A*02:01 YLIELIDRV (SEQ ID NO: 2250) TACE 250-258

A*02:01 FLAEDALNTV (SEQ ID NO: 2251) Epithelial Discoidin

Domain Receptor 1 (EDDR1) 867-876

A*02:01 GLMKYIGEV (SEQ ID NO: 2252) TRPM8 187-195

A*02:01 AILALLPAL (SEQ ID NO: 2253) Prostate Stem Cell Antigen

(PSCA) 105-133

A*02:01 GLQHWVPEL (SEQ ID NO: 2254) BA46 (Lactadherin) 97-106

A*02:01 GVRGRVEEI (SEQ ID NO: 2255) BCR-ABL

A*02:01 ITDQVPFSV (SEQ ID NO: 2256) gp100 (pmel) 209-217

A*02:01 KLCPVQLWV (SEQ ID NO: 2257) p53 139-147

A*02:01 KVAEELVHFL (SEQ ID NO: 2258) MAGEA3 112-120

(alternative version)

A*02:01 SLPPPGTRV (SEQ ID NO: 2259) p53 149-157

A*02:01 YLGSYGFRL (SEQ ID NO: 2260) p53 103-111

A*02:01 YLQLVFGIEV (SEQ ID NO: 2261) MAGEA2 157-166

A*02:01 TLQDIVYKL (SEQ ID NO: 2262) BMI1 74-82

A*02:01 YAIDLPVSV (SEQ ID NO: 2263) L-dopachrome tautomerase

488-496

A*02:01 AMVGAVLTA (SEQ ID NO: 2264) Tyrosinase 482-190

A*02:01 ATVGIMIGV (SEQ ID NO: 2265) CEACAM5 687-695

A*02:01 YVDPVITSI (SEQ ID NO: 2266) Hepatocyte growth factor

receptor 673-681

A*02:01 GVLLWEIFSL (SEQ ID NO: 2267) VEGFR1 28-37

A*02:01 LMAQEALAFL (SEQ ID NO: 2268)CAMEL 2-11

A*02:01 RVA(PHOSPHO-S)PTSGV (SEQ ID NO: 2269) Insulin receptor

substrate-2 1097-1105

A*02:01 RVASPTSGV (SEQ ID NO: 2270) IRS-2 1097-1105

A*02:01 ALNVYNGLL (SEQ ID NO: 2271) ACPP 299-307

A*02:01 ALSPVPPVV (SEQ ID NO: 2272) Bcl-2 85-93

A*02:01 ALVCYGPGI (SEQ ID NO: 2273) FAP alpha 463-471

A*02:01 ALWPWLLMAT (SEQ ID NO: 2274) RNF43 11-20

A*02:01 ALYLMELTM (SEQ ID NO: 2275) CB9L2

A*02:01 CLPSPSTPV (SEQ ID NO: 2276) BMI1 271-279

A*02:01 ELSDSLGPV (SEQ ID NO: 2277) PASD1 695-703

A*02:01 FLFLRNFSL (SEQ ID NO: 2278) TARP(V28L) 27-35

A*02:01 FLPSPLFFFL (SEQ ID NO: 2279) TARP(P5L) 5-13

A*02:01 GLFKCGIAV (SEQ ID NO: 2280) FAP 639-647

A*02:01 GLIQLVEGV (SEQ ID NO: 2281) TRAG-3 4-12

A*02:01 ILGVLTSLV (SEQ ID NO: 2282) DLK1 309-317

A*02:01 LLVPTCVFLV (SEQ ID NO: 2283) 691-700

A*02:01 MLAVFLPIV (SEQ ID NO: 2284) STEAP 292-300 (293L)

A*02:01 NLFETPVEA (SEQ ID NO: 2285) 194-202

A*02:01 QLGEQCWTV (SEQ ID NO: 2286) PSCA 44-51 (51A)

A*02:01 RLAEYQAYI (SEQ ID NO: 2287) SART3 309-317

A*02:01 SIDWFMVTV (SEQ ID NO: 2288) p31-39

A*02:01 SILLRDAGLV (SEQ ID NO: 2289) TRAG-3 57-66

A*02:01 SLFEPPPPG (SEQ ID NO: 2290) PSMA 85-93

A*02:01 SQADALKYV (SEQ ID NO: 2291) EZH2 729-737

A*02:01 WLSLKTLLSL (SEQ ID NO: 2292) Bcl-2 214-223

A*02:01 YLNRHLHTWI (SEQ ID NO: 2293) BCL-2 180-189

A*02:01 YLQWIEFSI (SEQ ID NO: 2294) Prominin1 744-752

A*02:01 YLYQWLGAPV (SEQ ID NO: 2295) Osteocalcin 51-60

A*02:01 KLMSSNSTDL (SEQ ID NO: 2296) HSP105 234-243

A*02:01 RLQGISPKI (SEQ ID NO: 2297) SSX2 103-111

A*02:01 AILALLPALL (SEQ ID NO: 2298) PSCA

A*02:01 ALIHHNTHL (SEQ ID NO: 2299) HER2 466-474

A*02:01 CMHLLLEAV (SEQ ID NO: 2300) MG50 624-632

A*02:01 FLIIWQNTM (SEQ ID NO: 2301) FSP26

A*02:01 FLPWHRLFLL (SEQ ID NO: 2302) Tyrosinase 207-216

A*02:01 FVWLHYYSV (SEQ ID NO: 2303) TRP2 185-193(L)

A*02:01 GLFGDIYLA (SEQ ID NO: 2304) CSNK1A1 26-34

A*02:01 GLFGDIYLAI (SEQ ID NO: 2305) CSNK1A1 26-35

A*02:01 ILLRDAGLV (SEQ ID NO: 2306) TRAG-3L 58-66

A*02:01 ILLVVVLGV (SEQ ID NO: 2307) Receptor tyrosine-protein

kinase erbB-2

A*02:01 ILNAMIAKI (SEQ ID NO: 2308) HAUS3 154-162

A*02:01 KASEYLQLV (SEQ ID NO: 2309) MAGEA2 153-161

A*02:01 KIWEELSVL (SEQ ID NO: 2310) MAGEA3 220-228

A*02:01 KLIDRTE(S)L (SEQ ID NO: 2311) LSP1 325-333

A*02:01 KLTGDENFTI (SEQ ID NO: 2312) Tyrosinase precursor

224-233

A*02:01 LLCYSCKAQV (SEQ ID NO: 2313) PSCA 17-26

A*02:01 LLLEAVPAV (SEQ ID NO: 2314) MG50 69-77

A*02:01 LLNQLQVNL (SEQ ID NO: 2315) Mucin2 467-475

A*02:01 LLRDAGLVKM (SEQ ID NO: 2316) TRAP 59-68

A*02:01 LLRRYNVAKV (SEQ ID NO: 2317) SOX11 266-275

A*02:01 LLSHGAVIEV (SEQ ID NO: 2318) Ankyrin NYBR1 158-167

A*02:01 LVFGIELMEV (SEQ ID NO: 2319) MAGEA3 160-169

A*02:01 LVFGIEVVEV (SEQ ID NO: 2320) MAGEA12 160-169

A*02:01 MLWGWREHV (SEQ ID NO: 2321) Mucin2 645-653

A*02:01 PLQPEQLQV (SEQ ID NO: 2322) Receptor tyrosine-protein

kinase erbB-2 437-445

A*02:01 QLMAFNHLI (SEQ ID NO: 2323) PAX3/FKHR 135-143

A*02:01 QLMPYGCLL (SEQ ID NO: 2324) Receptor tyrosine-protein

kinase erbB-2 845-853

A*02:01 RLGPTLMCL (SEQ ID NO: 2325) MG50 1244-1252

A*02:01 RLTRFLSRV (SEQ ID NO: 2326) CyclinD 228-236

A*02:01 RTF(S)PTYGL (SEQ ID NO: 2327) Desmuslin 426-434

A*02:01 SILLRDAGL (SEQ ID NO: 2328) TRAP 57-65

A*02:01 SLADEAEVYL (SEQ ID NO: 2329) GAS7 Neoepitope

A*02:01 SLDDYNHLV (SEQ ID NO: 2330) L-dopachrome tautomerase

288-296

A*02:01 SLYKFSPFPL (SEQ ID NO: 2331) O-linked

N-acetylglucosamine transferase FSP06

A*02:01 SMTR(S)PPRV (SEQ ID NO: 2332) SFRS2B 241-249

A*02:01 TLEEITGYL (SEQ ID NO: 2333) Receptor tyrosine-protein

kinase erbB-2 448-456

A*02:01 TLHCDCEIL (SEQ ID NO: 2334) MG50 210-218

A*02:01 VLEPPGARDV (SEQ ID NO: 2335) BIR 7 230-239

A*02:01 VLLALLMAGL (SEQ ID NO: 2336) Prostate stem cell

antigen 4-13

A*02:01 VLSVNVPDV (SEQ ID NO: 2337) MG50 625-633

A*02:01 VLVKSPNHV (SEQ ID NO: 2338) Receptor tyrosine-protein

kinase erbB-4 890-898

A*02:01 VMIG(S)PKKV (SEQ ID NO: 2339) Tensin3 1558-1566

A*02:01 VVLGVVFGI (SEQ ID NO: 2340) Receptor tyrosine-protein

kinase erbB-2 743-751

A*02:01 WLPKILGEV (SEQ ID NO: 2341) MG50 1051-1059

A*02:01 WLQYFPNPV (SEQ ID NO: 2342) Cytochrome P450 246-254

A*02:01 YLLDLSTNHL (SEQ ID NO: 2343) Fibromodulin 7-15

A*02:01 YLWWVNNQSL (SEQ ID NO: 2344) CEA 176-185

A*02:01 ALGGHPLLGV (SEQ ID NO: 2345) Dickkopf-related protein

1 20-29

A*02:01 ALLAGLVSL (SEQ ID NO: 2346) FGFR4 676-684

A*02:01 ALLTYMIAHI (SEQ ID NO: 2347) Thymidylate synthase

231-240

A*02:01 ALMDKSLHV (SEQ ID NO: 2348) MART-1 56-64

A*02:01 ALPPPLMLL (SEQ ID NO: 2349) Heparanase 8-16

A*02:01 ALSVMGVYV (SEQ ID NO: 2350) MAGEA9 223-231

A*02:01 ALVEFEDVL (SEQ ID NO: 2351) hnRNP L 140-148

A*02:01 ALWPWLLMA (SEQ ID NO: 2352) RNF43 11-19

A*02:01 AMLGTHTMEV (SEQ ID NO: 2353) Melanocyte-specific

secreted glycoprotein 184-193

A*02:01 AVIGALLAV (SEQ ID NO: 2354) Melanocyte-specific

secreted glycoprotein 20-28

A*02:01 CLYGNVEKV (SEQ ID NO: 2355) hnRNP L 404-412

A*02:01 DLIFGLNAL (SEQ ID NO: 2356) Heparanase 185-193

A*02:01 ELFQDLSQL (SEQ ID NO: 2357) ETV5 54-53

A*02:01 FAWERVRGL (SEQ ID NO: 2358) Cyclin-dependent kinase

inhibitor 1 97-105

A*02:01 FIASNGVKLV (SEQ ID NO: 2359) ACTN4 118-127 (K5N)

A*02:01 FLALIICNA (SEQ ID NO: 2360) Tubulin beta 4 283-291

A*02:01 FLDEFMEGV (SEQ ID NO: 2361) Malic enzyme 224-232

A*02:01 RMFPNAPYL (SEQ ID NO: 2362) WT-1 126-134 (Wilms tumor)

A*02:01 RLNMFTPYI (SEQ ID NO: 2363) Chlamydia trachomatis MOMP

258-266

A*02:01 NMFTPYIGV (SEQ ID NO: 2364) MOMP precursor 283-291

A*02:01 NLVPMVATV (SEQ ID NO: 2365) HCMV pp65 495-504

A*02:01 VLEETSVML (SEQ ID NO: 2366) HCMV IE1 316-324 (UL123)

A*02:01 VLAELVKQI (SEQ ID NO: 2367) HCMV IE1 81-89

A*02:01 MLNIPSINV (SEQ ID NO: 2368) pp65 120-128

A*02:01 LLLDRLNQL (SEQ ID NO: 2369) SARS-CoV Nucleocapsid

protein 223-231 (conserved in SARS-CoV-2)

A*02:01 FIAGLIAIV (SEQ ID NO: 2370) SARS-CoV-2 Spike

glycoprotein 1220-1228 (confirmed epitope)

A*02:01 ALNTLVKQL (SEQ ID NO: 2371) SARS-CoV Spike

glycoprotein precursor 940-948 (conserved in SARS-CoV-2)

A*02:01 LITGRLQSL (SEQ ID NO: 2372) SARS-CoV-2 Spike

glycoprotein 996-1004 (confirmed epitope)

A*02:01 NLNESLIDL (SEQ ID NO: 2373) SARS-CoV Spike

glycoprotein precursor 1174-1182 (conserved in SARS-Cov-2)

A*02:01 VLNDILSRL (SEQ ID NO: 2374) SARS-CoV Spike

glycoprotein precursor 958-966 (conserved in SARS-Cov-2)

A*02:01 YLQPRTFLL (SEQ ID NO: 2375) SARS-Cov-2 Spike

glycoprotein 269-277 (confirmed epitope)

A*02:01 LLYDANYFL (SEQ ID NO: 2376) SARS-CoV-2 ORF3a 139-147

(confirmed epitope)

A*02:01 RLQSLQTYV (SEQ ID NO: 2377) SARS-CoV-2 Spike

glycoprotein 1000-1008 (confirmed subdominant epitope)

A*02:01 KLWAQCVQL (SEQ ID NO: 2378) SARS-CoV-2 ORF1ab

3886-3894 (confirmed epitope)

A*02:01 TLYAVATTI (SEQ ID NO: 2379) Dengue NS4b 40-48

A*02:01 KLAEAIFKL (SEQ ID NO: 2380) Dengue NS5 563-571

A*02:01 ILIRTGLLVI (SEQ ID NO: 2381) Dengue NS2b 97-106

A*02:01 AIKRGLRTL (SEQ ID NO: 2382) Dengue NS3 112-120

A*02:01 LLLGLMILL (SEQ ID NO: 2383) Dengue NS4a 56-64

A*02:01 VLLLVTHYA (SEQ ID NO: 2384) Dengue NS4b 111-119

A*02:01 GLCTLVAML (SEQ ID NO: 2385) EBV BMLF-1 259-267

A*02:01 CLGGLLTMV (SEQ ID NO: 2386) EBV LMP-2 426-434

A*02:01 YLLEMLWRL (SEQ ID NO: 2387) EBV LMP-1 125-133

A*02:01 YLQQNWWTL (SEQ ID NO: 2388) EBV LMP1 159-167

A*02:01 YVLDHLIVV (SEQ ID NO: 2389) EBV BRLF1 109-117

A*02:01 FLYALALLL (SEQ ID NO: 2390) EBV LMP-2 356-364

A*02:01 TLDYKPLSV (SEQ ID NO: 2391) EBV BMRF1 208-216

A*02:01 LLDFVRFMGV (SEQ ID NO: 2392) EBV EBNA-3C 284-293

A*02:01 FLDKGTYTL (SEQ ID NO: 2393) EBV BALF-4 276-284

A*02:01 FLPSDFFPSV (SEQ ID NO: 2394) HBV core antigen 18-27

A*02:01 FLLTRILTI (SEQ ID NO: 2395) HBV envelope 183-191

A*02:01 GLSPTVWLSV (SEQ ID NO: 2396) HBV surface antigen

185-194

A*02:01 WLSLLVPFV (SEQ ID NO: 2397) HBV surface antigen 172-181

A*02:01 FLLSLGIHL (SEQ ID NO: 2398) HBV polymerase 573-581

A*02:01 FLPSDFFPSI (SEQ ID NO: 2399) HBV core 18-27 (subtype

ADR4)

A*02:01 VLHKRTLGL (SEQ ID NO: 2400) HBV X 92-100

A*02:01 GLSRYVARL (SEQ ID NO: 2401) HBV Pol 455-463

A*02:01 YMDDVVLGA (SEQ ID NO: 2402) HBV Polymerase 548-556

A*02:01 KLHLYSHPI (SEQ ID NO: 2403) HBV Pol 502-510

A*02:01 ELMTLATWV (SEQ ID NO: 2404) HBV core protein 64-72

A*02:01 DLMGYIPAV (SEQ ID NO: 2405) HCV core 132-140

A*02:01 CINGVCWTV (SEQ ID NO: 2406) HCV NS3 1073-1081

A*02:01 YLLPRRGPRL (SEQ ID NO: 2407) HCV core 35-44

A*02:01 VLSDFKTWL (SEQ ID NO: 2408) HCV NS5a 1987-1995

A*02:01 ALYDVVTKL (SEQ ID NO: 2409) HCV NS5b 2594-2602

A*02:01 KLVALGINAV (SEQ ID NO: 2410) HCV NS3 1406-1415

A*02:01 LLFNILGGWV (SEQ ID NO: 2411) HCV NS4b 1807-1816

A*02:01 KLSGLGINAV (SEQ ID NO: 2412) HCV NS3 1406-1415

A*02:01 DLMGYIPLV (SEQ ID NO: 2413) HCV core 132-140

A*02:01 CVNGVCWTV (SEQ ID NO: 2414) HCV NS3 1073-1081

A*02:01 GLQDCTMLV (SEQ ID NO: 2415) HCV NS5B 2727-2735

A*02:01 SLYNTVATL (SEQ ID NO: 2416) HIV-1 gag p17 76-84

A*02:01 ILKEPVHGV (SEQ ID NO: 2417) HIV-1 RT 476-484

A*02:01 TLNAWVKVV (SEQ ID NO: 2418) HIV-1 gag p24 19-27

A*02:01 KLTPLCVTL (SEQ ID NO: 2419) HIV-1 env gp120 90-98

A*02:01 GLADQLIHL (SEQ ID NO: 2420) HIV-1 vif 101-109

A*02:01 LTFGWCFKL (SEQ ID NO: 2421) HIV-1 nef 137-145

A*02:01 FLGKIWPS (SEQ ID NO: 2422) Gag 433-440

A*02:01 ALVEMGHHA (SEQ ID NO: 2423) HIV Vpu 66-74

A*02:01 RTLNAWVKV (SEQ ID NO: 2424) HIV gag 150-158

A*02:01 NVWATHACV (SEQ ID NO: 2425) HIV env gp 67-7

A*02:01 SLLNATAIAV (SEQ ID NO: 2426) HIV env 816-825

A*02:01 SLENTVATL (SEQ ID NO: 2427) HIV gag 77-85

A*02:01 SLVKHHMYI (SEQ ID NO: 2428) HIV vif 23-31

A*02:01 VIYHYVDDL (SEQ ID NO: 2429) HIV pol

A*02:01 YMLDLQPETT (SEQ ID NO: 2430) HPV 16 E7 11-20

A*02:01 KLPQLCTEL (SEQ ID NO: 2431) HPV 16 E6 18-26

A*02:01 YMLDLQPET (SEQ ID NO: 2432) HPV 16 E7 11-19

A*02:01 MLDLQPETT (SEQ ID NO: 2433) HPV 16 E7 12-20

A*02:01 VLMIKALEL (SEQ ID NO: 2434) Non muscle Myosin-9 741-749

A*02:01 QLFNHTMFI (SEQ ID NO: 2435) Non-muscle Myosin 478-486

A*02:01 QMARLAWEA (SEQ ID NO: 2436) 1116-1124

A*02:01 LLFGYPVYV (SEQ ID NO: 2437) Human T-cell lymphotropic

virus-1 (HTLV-1) tax

A*02:01 AVLDGLLSL (SEQ ID NO: 2438) HTLV bZIP factor 42-50

A*02:01 GLLSLEEEL (SEQ ID NO: 2439) bZIP factor 26-34

A*02:01 GILGFVFTL (SEQ ID NO: 2440) Influenza A MP 58-66

A*02:01 ILGFVFTLTV (SEQ ID NO: 2441) Influenza A MP 59-68

A*02:01 KLGEFYNQMM (SEQ ID NO: 2442) Flu BNP 85-94 (Influenza

B)

A*02:01 SITEVECFL (SEQ ID NO: 2443) VP1 36-44

A*02:01 ILMWEAVTL (SEQ ID NO: 2444) VP1 100-108

A*02:01 ALPHIIDEV (SEQ ID NO: 2445) LCMV envelope gp 10-18

A*02:01 YLVSIFLHL (SEQ ID NO: 2446) LCMV envelope gp 447-455

A*02:01 SLNQTVHSL (SEQ ID NO: 2447) NP 69-77

A*02:01 YLNKIQNSL (SEQ ID NO: 2448) Plasmodium falciparum CSP

334-342

A*02:01 FIDSYICQV (SEQ ID NO: 2449) miHAg H-Y (human SMCY)

311-319

A*02:01 YIGEVLVSV (SEQ ID NO: 2450) HA-2

A*02:01 VLHDDLLEA (SEQ ID NO: 2451) Minor Histocompatibility

Antigen HA-1 137-145

A*02:01 RTLDKVLEV (SEQ ID NO: 2452) miHAg HA-8

A*02:01 NEGATIVE (SEQ ID NO: 2453) Negative Control

A*02:01 TMFPHIIVDV (SEQ ID NO: 2454) Norovirus VP1 139-148

A*02:01 LLDVPTAAV (SEQ ID NO: 2455) Interferon gamma inducible

protein (GILT) 30 27-35

A*02:01 RILGAVAKV (SEQ ID NO: 2456) Vinculin 822-830

A*02:01 LMWYELSKI (SEQ ID NO: 2457) KSHVF-8 gB.492-500

A*02:01 ILEDIVLTL (SEQ ID NO: 2458) Streptococcus pyogenes

Cas9 615-623

A*02:01 KMLKEMGEV (SEQ ID NO: 2459) RSV NP 137-145

A*02:01 KLIANNTRV (SEQ ID NO: 2460) Mycobacterium bovis

antigen 85-A 200-208

A*02:01 GLPVEYLQV (SEQ ID NO: 2461) Mycobacterium bovis

antigen 85-A 6-14

A*02:01 GILTVSVAV (SEQ ID NO: 2462) 16 kDa

A*02:01 AMASTEGNV (SEQ ID NO: 2463) ESAT-6

A*02:01 VLTDGNPPEV (SEQ ID NO: 2464) 19 kDa

A*02:01 KVDDTFYYV (SEQ ID NO: 2465) Vaccinia virus Host range

protein 2 74-82

A*02:01 ILDDNLYKV (SEQ ID NO: 2466) Vaccinia virus Copenhagen

Protein G5 18-26

A*02:01 ALWALPHAA (SEQ ID NO: 2467) IE62 593-601

A*02:01 RLDDDGNFQL (SEQ ID NO: 2468) West Nile Virus NY-99

polyprotein precursor (1452-1461)

A*02:01 ATWAENIQV (SEQ ID NO: 2469) West Nile virus NY-99

polyprotein precursor 3390-3398

A*02:01 YTMDGEYRL (SEQ ID NO: 2470) West Nile virus NY-99

polyprotein precursor 2023-2031

A*02:01 SVGGVFTSV (SEQ ID NO: 2471) WNV envelope gp 430-438

A*02:01 SLFGQRIEV (SEQ ID NO: 2472) WNV nonstructural protein

4B 15-23

A*02:01 LLWNGPMAV (SEQ ID NO: 2473) NS4B 214-222

A*03:01 KQSSKALQR (SEQ ID NO: 2474) bcr-abl 210 kD fusion

protein 21-29

A*03:01 ALLAVGATK (SEQ ID NO: 2475) gp100 (pmel17) 17-25

A*03:01 ATGFKQSSK (SEQ ID NO: 2476) bcr-abl 210 kD fusion

protein 259-269

A*03:01 RISTFKNWPK (SEQ ID NO: 2477) Survivin-3A 18-27 (27K)

A*03:01 RLGLQVRKNK (SEQ ID NO: 2478) RhoC 176-185 (177L)

A*03:01 RLLFFAPTR (SEQ ID NO: 2479) Mcl-1 95-103

A*03:01 QVLKKIAQK (SEQ ID NO: 2480) HMOX1 145-153

A*03:01 RIAAWMATY (SEQ ID NO: 2481) 165-173

A*03:01 KLGGALQAK (SEQ ID NO: 2482) HCMV IE1 184-192

A*03:01 KTFPPTEPK (SEQ ID NO: 2483) SARS-CoV-2 Nucleocapsid

protein 362-370 (confirmed epitope)

A*03:01 ELERAADVK (SEQ ID NO: 2484) Dengue NS2b 52-60

A*03:01 RVSTVQQLTK (SEQ ID NO: 2485) Dengue C 22-31

A*03:01 RIEPSWADVK (SEQ ID NO: 2486) Dengue NS3 64-74

A*03:01 RVIDPRRCMK (SEQ ID NO: 2487) Dengue NS3 422-431

A*03:01 KITAEWLWK (SEQ ID NO: 2488) Dengue NS5 375-383

A*03:01 RLRAEAQVK (SEQ ID NO: 2489) EBV EBNA 3A 603-611

A*03:01 RVRAYTYSK (SEQ ID NO: 2490) EBV BRLF1

A*03:01 RVCEKMALY (SEQ ID NO: 2491) HCV NS5B 2588-2596

A*03:01 QVPLRPMTYK (SEQ ID NO: 2492) HIV-1 nef 73-82

A*03:01 RLRPGGKKK (SEQ ID NO: 2493) HIV-1 gag p17 19-27

A*03:01 AIFQSSMTK (SEQ ID NO: 2494) HIV pol 325-333

A*03:01 KLCLRFLSK (SEQ ID NO: 2495) HPV 33 E6 64-72

A*03:01 ILRGSVAHK (SEQ ID NO: 2496) Influenza A (PR8) NP

265-274

A*11:01 KTFPPTEPK (SEQ ID NO: 2483) SARS-CoV-2 Nucleocapsid

protein 362-370 (confirmed epitope)

A*11:01 GTSGSPIINR (SEQ ID NO: 2497) Dengue NS3 serotype 3&4

133-142

A*11:01 GTSGSPIIDK (SEQ ID NO: 2498) Dengue NS3 133-142

A*11:01 GTSGSPIVNR (SEQ ID NO: 2499) NS3 serotype 1 133-142

A*11:01 GTSGSPIVDR (SEQ ID NO: 2500) Dengue NS3 serotype 2

133-142

A*11:01 GTSGSPIADK (SEQ ID NO: 2501) Dengue NS3 133-142

A*11:01 RVSTVQQLTK (SEQ ID NO: 2485) Dengue C 22-31

A*11:01 RIEPSWADVK (SEQ ID NO: 2486) Dengue NS3 64-74

A*11:01 RVIDPRRCMK (SEQ ID NO: 2487) Dengue NS3 422-431

A*11:01 KITAEWLWK (SEQ ID NO: 2488) Dengue NS5 375-383

A*11:01 IVTDFSVIK (SEQ ID NO: 2502) EBV EBNA-4 416-424

A*11:01 SSCSSCPLSK (SEQ ID NO: 2503) EBV LMP-2 340-349

A*11:01 ATIGTAMYK (SEQ ID NO: 2504) EBV BRLF1 134-142

A*11:01 AVFDRKSDAK (SEQ ID NO: 2505) EBNA3B 399-408

A*11:01 YVNVNMGLK (SEQ ID NO: 2506) HBV core antigen 88-96

A*11:01 YVNTNMGLK (SEQ ID NO: 2507) HBV core 88-96

A*11:01 STLPETTVVRR (SEQ ID NO: 2508) HBV core 141-151

A*11:01 AVDLSHFLK (SEQ ID NO: 2509) HIV nef 84-92

A*11:01 ACQGVGGPGHK (SEQ ID NO: 2510) HIV gag p24

A*11:01 NTLEQTVKK (SEQ ID NO: 2511) HPV 33 E6 86-94

A*11:01 SIIPSGPLK (SEQ ID NO: 2512) Influenza A MP 13-21

A*11:01 RMVLASTTAK (SEQ ID NO: 2513) Influenza A MP1 178-187

A*11:01 KSMREEYRK (SEQ ID NO: 2514) Influenza A MP2 70-78

A*24:02 TYFSLNNKF (SEQ ID NO: 2515) Adenovirus 5 Hexon 37-45

A*24:02 TYACFVSNL (SEQ ID NO: 2516) Carcinogenic Embryonic

Antigen (CEA) 652-660

A*24:02 AFLPWHRLF (SEQ ID NO: 2517) Tyrosinase 188-196

A*24:02 IMPKAGLLI (SEQ ID NO: 2518) MAGE-A3

A*24:02 VYFFLPDHL (SEQ ID NO: 2519) gp100-intron 4 (170-178)

A*24:02 EYLQLVFGI (SEQ ID NO: 2520) MAGEA2 156-164

A*24:02 TYLPTNASL (SEQ ID NO: 2521) HER-2/neu 63-71

A*24:02 VYGFVRACL (SEQ ID NO: 2522) Telomerase reverse

transcriptase (hTRT) 461-469

A*24:02 TFPDLESEF (SEQ ID NO: 2523) MAGEA3 97-105

A*24:02 DYLQYVLQI (SEQ ID NO: 2524) MiHA ACC1 15-23

A*24:02 RYCNLEGPPI (SEQ ID NO: 2525) Lymphocyte antigen 6

complex locus K (LY6K) 177-186

A*24:02 AYACNTSTL (SEQ ID NO: 2526) Survivin 80-88

A*24:02 CYASGWGSI (SEQ ID NO: 2527) Prostate Specific

Antigen-1 153-161

A*24:02 DYLNEWGSRF (SEQ ID NO: 2528) CDH3 807-816

A*24:02 EYCPGGNLF (SEQ ID NO: 2529) MELK 87-95 (93N)

A*24:02 EYYELFVNI (SEQ ID NO: 2530) DEP DC1 294-302

A*24:02 GYCTQIGIF (SEQ ID NO: 2531) HENMT1 221-229

A*24:02 IYTWIEDHF (SEQ ID NO: 2532) FOXM1 262-270

A*24:02 NYQPVWLCL (SEQ ID NO: 2533) RNF43 721-729 (722Y)

A*24:02 RYNAQCQETI (SEQ ID NO: 2534) Midkine 110-119

A*24:02 EYRALQLHL (SEQ ID NO: 2535) CA9 219-227

A*24:02 SYRNEIAYL (SEQ ID NO: 2536) TTK protein kinase 551-559

A*24:02 VYLRVRPLL (SEQ ID NO: 2537) KIF20A 67-75

A*24:02 VYYNWQYLL (SEQ ID NO: 2538) IL13r 146-154

A*24:02 VYALPLKML (SEQ ID NO: 2539) HCMV pp65 113-121

A*24:02 QYDPVAALF (SEQ ID NO: 2540) HCMV pp65 341-349

A*24:02 AYAQKIFKI (SEQ ID NO: 2541) CMV IE-1 248-256

A*24:02 QYSDRRWCF (SEQ ID NO: 2542) Dengue NS3 557-565

(Singapore/S275/1990)

A*24:02 TYGPVFMCL (SEQ ID NO: 2543) EBV LMP-2 419-427

A*24:02 PYLFWLAAI (SEQ ID NO: 2544) EBV LMP2 131-139

A*24:02 TYGPVFMSL (SEQ ID NO: 2545) EBV LMP2 419-427

A*24:02 EYLVSFGVW (SEQ ID NO: 2546) HBV core 117-125

A*24:02 KYTSFPWLL (SEQ ID NO: 2547) HBV polymerase 756-764

A*24:02 FFPSIRDLL (SEQ ID NO: 2548) HBV core protein 23-31

A*24:02 AYSQQTRGL (SEQ ID NO: 2549) HCV NS3 1031-1039

A*24:02 RYPLTFGWCY (SEQ ID NO: 2550) HIV-1 Nef 134-143

A*24:02 RYLKDQQLL (SEQ ID NO: 2551) HIV-1 gag gp41 67-75

A*24:02 RYLRDQQLL (SEQ ID NO: 2552) HIV env

A*24:02 RYPLTFGWCF (SEQ ID NO: 2553) HIV nef 143-152

A*24:02 RYPLTFGW (SEQ ID NO: 2554) HIV nef

A*24:02 VYDFAFRDL (SEQ ID NO: 2555) HPV16 E6

A*24:02 SFHSLHLLF (SEQ ID NO: 2556) HTLV Tax 301-309

A*29:02 KEKYIDQEEL (SEQ ID NO: 2557) HSP90 alpha 280-288

(Pathologic Conditions)

A*29:02 LYNTVATLY (SEQ ID NO: 2558) HIV gag 79-86

A*29:02 SFDPIPIHY (SEQ ID NO: 2559) HIV env 216-224

A*29:02 SFNCRGEFFY (SEQ ID NO: 2560) HIV env 382-391

A*68:01 TVSGNILTIR (SEQ ID NO: 2561) NY-ESO-1 127-136

B*07:02 VPQYGYLTL (SEQ ID NO: 2562) AAV2 372-380

B*07:02 KPYSGTAYNSL (SEQ ID NO: 2563) Adenovirus Hexon 114-124

B*07:02 KPYSGTAYNAL (SEQ ID NO: 2564) Adenovirus Hexon 114-124

B*07:02 LPLMRKAYL (SEQ ID NO: 2565) LT antigen 27-35

B*07:02 LPWHRLFLL (SEQ ID NO: 2566) Tyrosinase 208-216

B*07:02 EPR(PHOSPHO-S)PSHSM (SEQ ID NO: 2567) Insulin receptor

substrate 2

B*07:02 TPNQRQNVC (SEQ ID NO: 2568) P2X5

B*07:02 APRGVRMAV (SEQ ID NO: 2569) LAGE-1 46-54

B*07:02 LPVSPRLQL (SEQ ID NO: 2570) CEACAM 185-193

B*07:02 TPRVTGGGAM (SEQ ID NO: 2571) HCMV pp65 417-426

B*07:02 RPHERNGFTVL (SEQ ID NO: 2572) HCMV pp65 265-275

B*07:02 SPRWYFYYL (SEQ ID NO: 2573) SARS-CoV-2 Nucleocapsid

protein 105-113 (confirmed epitope)

B*07:02 APTRVVAAEM (SEQ ID NO: 2574) Dengue NS3 serotype 2

222-231

B*07:02 RPPIFIRRL (SEQ ID NO: 2575) EBV EBNA-3A 247-255

B*07:02 RPQGGSRPEFVKL (SEQ ID NO: 2576) EBV BMRF1 116-128

B*07:02 QPRAPIRPI (SEQ ID NO: 2577) EBV EBNA-3C 881-889

B*07:02 LPSDFFPSV (SEQ ID NO: 2578) HBV core 19-27

B*07:02 GPRLGVRAT (SEQ ID NO: 2579) HCV core 41-49

B*07:02 DPRRRSRNL (SEQ ID NO: 2580) HCV core 111-119

B*07:02 IPRRIRQGL (SEQ ID NO: 2581) HIV-1 env gp120 848-856

B*07:02 TPGPGVRYPL (SEQ ID NO: 2582) HIV-1 nef 128-137

B*07:02 GPGHKARVL (SEQ ID NO: 2583) HIV gag p24 223-231

B*07:02 KPTLKEYVL (SEQ ID NO: 2584) HPV 33 E7 5-13

B*07:02 LPVSCPEDL (SEQ ID NO: 2585) bZIP factor 10-18

B*07:02 QPEWFRNVL (SEQ ID NO: 2586) Influenza A PB1 329-337

B*07:02 SPIVPSFDM (SEQ ID NO: 2587) Influenza A NP 473-481

B*07:02 SPSVDKARAEL (SEQ ID NO: 2588) MiHAg SMCY 1041-1051

B*08:01 GFKQSSKAL (SEQ ID NO: 2589) bcr-abl 210 kD fusion

protein 19-27

B*08:01 ELRRKMMYM (SEQ ID NO: 2590) IE1 199-207

B*08:01 ELKRKMIYM (SEQ ID NO: 2591) CMV IE-1 199-207 (R/L

Position 3, M/I Position 7)

B*08:01 RIKQKGIL (SEQ ID NO: 2592) Dengue NS3 25-32

B*08:01 LEKTKKDL (SEQ ID NO: 2593) Dengue NS4a 6-13

B*08:01 FLRGRAYGL (SEQ ID NO: 2594) EBV EBNA-3A 193-201

B*08:01 RAKFKQLL (SEQ ID NO: 2595) EBV BZLF-1 190-197

B*08:01 QAKWRLQTL (SEQ ID NO: 2596) EBV EBNA3A 158-166

B*08:01 GLKILQLL (SEQ ID NO: 2597) HBV external core Ag

B*08:01 HSKKKCDEL (SEQ ID NO: 2598) HCV NS3 1395-1403

B*08:01 FLKEKGGL (SEQ ID NO: 2599) HIV-1 nef 90-97

B*08:01 GEIYKRWII (SEQ ID NO: 2600) HIV-1 gag p24 261-269

B*08:01 EIYKRWII (SEQ ID NO: 2601) HIV p24 gag 128-135

B*08:01 YLKDQQLL (SEQ ID NO: 2602) Env 586-593

B*15:01 CLIPTAMAF (SEQ ID NO: 2603) Dengue C 107-115

B*15:01 RLRPGGKKKY (SEQ ID NO: 2604) HIV-1 p17 20-29

B*27:05 GREGLATEK (SEQ ID NO: 2605) BRAF 594-601 (600E)

B*27:05 GRFGLATVK (SEQ ID NO: 2606) BRAF 594-601 (600V)

B*27:05 RMFPNAPYL (SEQ ID NO: 2362) WT-1 126-134 (Wilms tumor)

B*27:05 ARKLLLDNL (SEQ ID NO: 2607) PqqC-like protein 70-78

B*27:05 NRAKQVIKL (SEQ ID NO: 2608) Probable ATP-dependent Clp

protease ATP-binding subunit 7-15

B*27:05 QRNAPRITF (SEQ ID NO: 2609) SARS-CoV-2 Nucleocapsid

protein 9-17 (confirmed epitope)

B*27:05 ARMILMTHF (SEQ ID NO: 2610) HCV NS5B 2841-2849

B*27:05 KRWIILGLNK (SEQ ID NO: 2611) HIV-1 gag p24 265-274

B*27:05 KRWIIMGLNK (SEQ ID NO: 2612) HIV-1 Gag p24 263-272

B*27:05 GRAFVTIGK (SEQ ID NO: 2613) HIV-1 gp100 103-111

B*27:05 SRYWAIRTR (SEQ ID NO: 2614) Influenza A NP 383-391

B*35:01 IPYLDGTFY (SEQ ID NO: 2615) Adenovirus Hexon

B*35:01 MPFATPMEA (SEQ ID NO: 2616) NY-ESO-1 94-102

B*35:01 IPSINVHHY (SEQ ID NO: 2617) HCMV pp65 123-131

B*35:01 TPEGIIPTL (SEQ ID NO: 2618) Dengue NS3 500-508

B*35:01 YPLHEQHGM (SEQ ID NO: 2619) EBV EBNA-3A 458-466

B*35:01 EPLPQGQLTAY (SEQ ID NO: 2620) EBV BZLF-1 54-64

B*35:01 HPVAEADYFEY (SEQ ID NO: 2621) EBV EBNA-1 407-417(4A)

B*35:01 HPNIEEVAL (SEQ ID NO: 2622) HCV NS3 1359-1367

B*35:01 CPNSSIVY (SEQ ID NO: 2623) HCV E1 207-214

B*35:01 NPDIVIYQY (SEQ ID NO: 2624) HIV-1 RT 330-338

B*35:01 NPDIVIYQY (SEQ ID NO: 2625) HIV-1 HIV-1 RT 328-336

B*35:01 VPLDEDFRKY (SEQ ID NO: 2626) HIV-1 HIV-1 RT 273-282

B*40:01 MEVTPSGTWL (SEQ ID NO: 2627) SARS-CoV-2 Nucleocapsid

protein 322-330 (confirmed epitope)

B*40:01 GEARKTFVEL (SEQ ID NO: 2628) Dengue NS3 528-537

B*40:01 IEDPPENSL (SEQ ID NO: 2629) EBV LMP2 200-208

B*40:01 REISVPAEIL (SEQ ID NO: 2630) HCV NS5a 2266-2275

B*40:01 KEKGGLEGL (SEQ ID NO: 2631) HIV-1 Nef 92-100

C*06:02 TRATKMQVI (SEQ ID NO: 2632) pp65 211-219

C*06:02 QIKVRVDM (SEQ ID NO: 2633) IE1 88-95

C*06:02 TRRFLPQIL (SEQ ID NO: 2634) NS3 205-213

C*07:02 CRVLCCYVL (SEQ ID NO: 2635) IE1 309-317

E*01:01 VMAPRTLIL (SEQ ID NO: 2636) HLA-C leader sequence

peptide

E*01:01 VMAPRTLVL (SEQ ID NO: 2637) HLA-A leader sequence

peptide

DRB1*01:01 DSVTPMILKAQKGGNL (SEQ ID NO: 2641) Dog dander Can

f 1 33-48

DRB1*01:01 KCIEWEKAQHGA (SEQ ID NO: 2642) Mugwort pollen Art

v 1 25-36

DRB1*01:01 LPVVLENARILKNCVDAK (SEQ ID NO: 2643) Cat dander

Fel d 1 53-70

DRB1*01:01 LRQMRTVTPIRMQGG (SEQ ID NO: 2644) House dust mite

Der p1 96-110

DRB1*01:01 VAAAPQVKYAVFEAALTKAI (SEQ ID NO: 2645) Timothy

grass Phl p 5 p26

DRB1*01:01 YESYKFIPALEAAVK (SEQ ID NO: 2646) Grass pollen

allergen Phl p 5 196-210

DRB1*01:01 ETLLRAVESYLLAHS (SEQ ID NO: 2647) Birch pollen

allergen Bet v 1 142-156

DRB1*01:01 ACYEFLWGPRALVETS (SEQ ID NO: 2648) MAGE-A3 267-282

DRB1*01:01 LKEFTVSGNILTIRL (SEQ ID NO: 2649) NY-ESO-1 123-137

DRB1*01:01 LLEFYLAMPFATPME (SEQ ID NO: 2650) NY-ESO-1 87-101

DRB1*01:01 KVPIKWMALESILRRRF (SEQ ID NO: 2651) HER2 883-899

DRB1*01:01 LPLKMLNIPSINVH (SEQ ID NO: 2652) CMV pp65 116-129

DRB1*01:01 GAALQIPFAMQMAYRF (SEQ ID NO: 2653) SARS-CoV S

protein (873-888) (conserved in SARS-CoV-2)

DRB1*01:01 MAYRFNGIGVTQNVLY (SEQ ID NO: 2654) SARS-CoV S

protein (884-899) (conserved in SARS-CoV-2)

DRB1*01:01 QLIRAAEIRASANLAATK (SEQ ID NO: 2655) SARS-CoV S

protein (993-1010) (conserved in SARS-CoV-2)

DRB1*01:01 TVFYNIPPMPL (SEQ ID NO: 2656) EBV EBNA2 280-290

DRB1*01:01 TSLYNLRRGTALA (SEQ ID NO: 2657) EBV EBNA1 515-527

DRB1*01:01 QAGFFLLTRILTIPQS (SEQ ID NO: 2658) HBV envelope

179-194

DRB1*01:01 PPAYRPPNAPILSTL (SEQ ID NO: 2659) HBV core 158-172

DRB1*01:01 LCWGELMTLATWVGVN (SEQ ID NO: 2660) HBV core 60-75

DRB1*01:01 TLLFNILGGWVAA (SEQ ID NO: 2661) HCV polyprotein

1806?1818, NS4b

DRB1*01:01 DYVDRFYKTLRAE (SEQ ID NO: 2662) HIV-1 gag 295-307

DRB1*01:01 KRWIILGLNKIVRMYSPTSI (SEQ ID NO: 2663) HIV-1 gag

263-282

DRB1*01:01 FRDYVDRFYKTLRAEQASQE (SEQ ID NO: 2664) HIV-1 gag

293-312

DRB1*01:01 SGPLKAEIAQRLEDV (SEQ ID NO: 2665) Influenza A MP

17-31

DRB1*01:01 PKYVKQNTLKLAT (SEQ ID NO: 2666) Influenza A HA

307-319

DRB1*01:01 EYLNKIQNSLSTEWSPCSVT (SEQ ID NO: 2667) CSP 326-345

DRB1*01:01 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*01:01 QEIYMQHTYPISA (SEQ ID NO: 2669) TT 257-269

DRB1*01:01 GEEYLILSARDVLAV (SEQ ID NO: 2670) Mtb 10 kDa

chaperonin GroES

DRB1*03:01 HTYTIDWTKDAVTWS (SEQ ID NO: 2671) Aspergillus

fumigatus Crf1/p41 171-185

DRB1*03:01 VYYLTRDPTTPLARAA (SEQ ID NO: 2672) HCV polyprotein

2800-2815, NS5b

DRB1*03:01 PIVQLQGDSNCLKCFR (SEQ ID NO: 2673) HPV E2 285-300

DRB1*03:01 MEAIAKRLDACQDQLLELYE (SEQ ID NO: 2674) HPV E2 1-20

DRB1*03:01 RQIFGDYKTTIC (SEQ ID NO: 2675) PLP 98-109

DRB1*03:01 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*03:01 MAKTIAYDEEARR (SEQ ID NO: 2676) Mtb groEL 1-13

DRB1*03:01 KTIAYDEEARR (SEQ ID NO: 2677) Mtb groEL 3-13

DRB1*04:01 APYHFDLSGHAFG (SEQ ID NO: 2678) Rye grass Lol p1

124-136

DRB1*04:01 ELEKYQQLNSERGVPN (SEQ ID NO: 2679) Cow dander Bos

d 2 143-158

DRB1*04:01 ETLLRAVESYLLAHS (SEQ ID NO: 2680) Birch pollen

allergen Bet v 1 141-155

DRB1*04:01 VAAAPQVKYAVFEAALTKAI (SEQ ID NO: 2645) Timothy

grass Phl p 5 p26

DRB1*04:01 FHTYTIDWTKDAVTW (SEQ ID NO: 2681) Aspergillus

fumigatus Crf1/p41 170-184

DRB1*04:01 NFIRMVISNPAAT (SEQ ID NO: 2682) GAD65 555-567

DRB1*04:01 IAFTSEHSHFSLK (SEQ ID NO: 2683) GAD65 274-286

DRB1*04:01 GAGSLQPLALEGSLQKRG (SEQ ID NO: 2684) Proinsulin

73-90

DRB1*04:01 GIVEQCCTSICSLYQ (SEQ ID NO: 2685) Proinsulin 90-104

DRB1*04:01 ATEG[cit]V[cit]VNSAYQDK (SEQ ID NO: 2686) Cit

aggregan 89-103

DRB1*04:01 ATIKAEFV[cit]AETPYM (SEQ ID NO: 2687) Cit CILP

297?311

DRB1*04:01 DVMNILLQYVVKSFDRSTKV (SEQ ID NO: 2030) GAD65 113-132

DRB1*04:01 GKLYGI[cit]DV[cit]STRDR (SEQ ID NO: 2688) Cit CILP

982?996

DRB1*04:01 GVYAT[cit]SSAV[cit]L[cit]SSVPGVR (SEQ ID NO: 2689)

Cit vimentin aa 59-78

DRB1*04:01 IFDS[cit]GNPTVEVDLF (SEQ ID NO: 2690) Cit alpha-

enolase 11-25

DRB1*04:01 KGMAALPRLIAFTSEHSHFS (SEQ ID NO: 2691) GAD65 265?284

DRB1*04:01 QDFTN[cit]INKLKNS (SEQ ID NO: 2692) Cit fibrinogen-

alpha 79-91

DRB1*04:01 WNRQLYPEWTEAQRLD (SEQ ID NO: 2693) gp100 44-59

DRB1*04:01 ISPNSVFSQWRVVCDSLEDYD (SEQ ID NO: 2694) Tyrp1

277-297

DRB1*04:01 ESEFQAALSRKVAKL (SEQ ID NO: 2695) MAGE-A6 102-116

DRB1*04:01 LTQYFVQENYLEYRQVPG (SEQ ID NO: 2696) MAGE-A6 246-263

DRB1*04:01 YACFVSNLATGRNNS (SEQ ID NO: 2697) CEA 653-667

DRB1*04:01 NYTLRVDCTPLMYSL (SEQ ID NO: 2698) PSMA 459-473

DRB1*04:01 IYRRRLMKQDFSVPQLPHS (SEQ ID NO: 2699) gp100 615-633

DRB1*04:01 ALHIYMDGTMSQVQGSA (SEQ ID NO: 2700) tyrosinase

365-381

DRB1*04:01 RNGYRALMDKSLHVGTQCALTRR (SEQ ID NO: 2701) MART-1

51-73

DRB1*04:01 LKEFTVSGNILTIRL (SEQ ID NO: 2649) NY-ESO-1 123-137

DRB1*04:01 LLEFYLAMPFATPME (SEQ ID NO: 2650) NY-ESO-1 87-101

DRB1*04:01 KVPIKWMALESILRRRF (SEQ ID NO: 2651) HER2 883-899

DRB1*04:01 QALNTLVKQLSSNFGAI (SEQ ID NO: 2702) SARS-CoV S

protein (939-955) (conserved in SARS-CoV-2)

DRB1*04:01 QLIRAAEIRASANLAATK (SEQ ID NO: 2655) SARS-CoV S

protein (993-1010) (conserved in SARS-CoV-2)

DRB1*04:01 PYYVVDLSVRGM (SEQ ID NO: 2703) EBV BHRF1 122-133

DRB1*04:01 AEGLRALLARSHVER (SEQ ID NO: 2704) EBV EBNA1 482-496

DRB1*04:01 GQTYHLIVDTDSLGNPSLSV (SEQ ID NO: 2705) EBV EBNA2

11-30

DRB1*04:01 LSFLPSDFFPSVRDL (SEQ ID NO: 2706) HBV core 45-59

DRB1*04:01 GYKVLVLNPSVAATL (SEQ ID NO: 2707) HCV polyprotein

1248-1262, NS3

DRB1*04:01 SGIQYLAGLSTLPGNPAIASL (SEQ ID NO: 2708) HCV

polyprotein 1770-1790, NS4b

DRB1*04:01 AFSPEVIPMFSALSEGATPQ (SEQ ID NO: 2709) HIV-1 gag

163-182

DRB1*04:01 FWRGENGRKTRIAYERMCNILKGK (SEQ ID NO: 2710)

Influenza A NP 206-229

DRB1*04:01 GFVFTLTVPSER (SEQ ID NO: 2711) Influenza A MP 61-72

DRB1*04:01 PKYVKQNTLKLAT (SEQ ID NO: 2666) Influenza A HA

307-319

DRB1*04:01 EYLNKIQNSLSTEWSPCSVT (SEQ ID NO: 2667) CSP 326-345

DRB1*04:01 RTELLKDAIGEGK (SEQ ID NO: 2712) MOG 97-109

DRB1*04:01 TWTTCQSIAFPSKTSASIGS (SEQ ID NO: 2713) PLP 180-199

DRB1*04:01 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*04:01 SSPKPWIYATSNLAS (SEQ ID NO: 2714) Rituximab Light

chain 41-55

DRB1*04:02 PKYVKQNTLKLAT (SEQ ID NO: 2666) Influenza A HA

307-319

DRB1*04:02 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*04:05 LWWVNNQSLPVSP (SEQ ID NO: 2715) CEA 321-333ÿ

DRB1*04:05 PKYVKQNTLKLAT (SEQ ID NO: 2666) Influenza A HA

307-319

DRB1*04:05 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*07:01 DRVNFKYSFSVIE (SEQ ID NO: 2716) Alder pollen Aln g

1 76-88

DRB1*07:01 VAAAPQVKYAVFEAALTKAI (SEQ ID NO: 2645) Timothy

grass Phl p 5 p26

DRB1*07:01 VGLLKAVESYLLA (SEQ ID NO: 2717) Alder pollen Aln g

1 142-154

DRB1*07:01 YYSNVTATRLLSSTNS (SEQ ID NO: 2718) Pertussis toxin

subunit 2 129-144

DRB1*07:01 EPDVYYTSAFVFPTK (SEQ ID NO: 2719) CMV pp65 177-191

DRB1*07:01 PDDYSNTHSTRYVTV (SEQ ID NO: 2720) CMV gB 215-229

DRB1*07:01 VPGLYSPCRAFFNKEELL (SEQ ID NO: 2721) EBV MCP

1264-1281

DRB1*07:01 PGPLRESIVCYFMVFLQTHI (SEQ ID NO: 2722) EBV EBNA1

551-570

DRB1*07:01 VYGGSKTSLYNLRRGTALAI (SEQ ID NO: 2723) EBV EBNA1

509-528

DRB1*07:01 PRSPTVFYNIPPMPLPPSQL (SEQ ID NO: 2724) EBV EBNA2

276-295

DRB1*07:01 AYCLWMMLLISQAEAALELIT (SEQ ID NO: 2725) HCV NS2

732-753

DRB1*07:01 WPLLLLLLALPQRAYAQ (SEQ ID NO: 2726) HCV NS2 47-63

DRB1*07:01 SLTITSLLRRHNWITSCS (SEQ ID NO: 2727) HCV NS5a

1957-1975

DRB1*07:01 TTVRLRAYMNTPGLPVC (SEQ ID NO: 2728) HCV NS3

1535-1551

DRB1*07:01 FRDYVDRFYKTLRAEQASQE (SEQ ID NO: 2664) HIV-1 gag

293-312

DRB1*07:01 PKYVKQNTLKLAT (SEQ ID NO: 2666) Influenza A HA

307-319

DRB1*07:01 EYLNKIQNSLSTEWSPCSVT (SEQ ID NO: 2667) CSP 326-345

DRB1*07:01 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*07:01 WVKQTPGRGLEWIGA (SEQ ID NO: 2729) Rituximab Heavy

chain 36-50

DRB1*07:01 SSPKPWIYATSNLAS (SEQ ID NO: 2714) Rituximab Light

chain 41-55

DRB1*07:01 EWVAEIRSKSINSAT (SEQ ID NO: 2730) Infliximab Heavy

chain 46-60

DRB1*09:01 VAAAPQVKYAVFEAALTKAI (SEQ ID NO: 2645) Timothy

grass Phl p 5 p26

DRB1*09:01 LLEFYLAMPFATPME (SEQ ID NO: 2650) NY-ESO-1 87-101

DRB1*09:01 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*09:01 MTEQQWNFAGIEAAA (SEQ ID NO: 2731) Mtb ESAT6 1-15

DRB1*11:01 LEAAFNAEFNEIRRVLLEEN (SEQ ID NO: 2732) Peanut Ara

h 1 321-340

DRB1*11:01 TSRNNPFYFPSRRFSTRYGN (SEQ ID NO: 2733) Peanut Ara

h 1 169-188

DRB1*11:01 VVNKGTGNLELVAVRKEQQQ (SEQ ID NO: 2734) Peanut Ara

h 1 457-476

DRB1*11:01 VAAAPQVKYAVFEAALTKAI (SEQ ID NO: 2645) Timothy

grass Phl p 5 p26

DRB1*11:01 TSYVKVLHHMVKISG (SEQ ID NO: 2735) MAGE-A3 281-295

DRB1*11:01 ENIQRFLPNPAGVQLEDPEF (SEQ ID NO: 2736) Factor VIII

589?608

DRB1*11:01 FRDYVDRFYKTLRAEQASQE (SEQ ID NO: 2664) HIV-1 gag

293-312

DRB1*11:01 PKYVKQNTLKLAT (SEQ ID NO: 2666) Influenza A HA

307-319

DRB1*11:01 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*11:01 VSIDKFRIFCKALNPK (SEQ ID NO: 2737) TT 1084-1099

DRB1*11:01 FNNFTVSFWLRVPKVSASHLE (SEQ ID NO: 2738) TT 947-967

DRB1*11:01 KFIIKRYTPNNEIDSF (SEQ ID NO: 2739) TT 1174-1189

DRB1*11:01 QYIKANSKFIGITEL (SEQ ID NO: 2740) TT 830-844

DRB1*11:01 WVKQTPGRGLEWIGA (SEQ ID NO: 2729) Rituximab Heavy

chain 36-50

DRB1*15:01 VAAAPQVKYAVFEAALTKAI (SEQ ID NO: 2645) Timothy

grass Phl p 5 p26

DRB1*15:01 VGLLKAVESYLLA (SEQ ID NO: 2717) Alder pollen Aln

g 1 142-154

DRB1*15:01 AVNIVGYSNAQGVDY (SEQ ID NO: 2741) House dust mite

Der p1 251-265

DRB1*15:01 ETLLRAVESYLLAHS (SEQ ID NO: 2647) Birch pollen

allergen Bet v 1 142-156

DRB1*15:01 LRQMRTVTPIRMQGG (SEQ ID NO: 2644) House dust mite

Der p1 96-110

DRB1*15:01 DENPVVHFFKNIVTPRTPP (SEQ ID NO: 2027) Myelin basic

protein 83-101

DRB1*15:01 MSIYVYALPLKMLNI (SEQ ID NO: 2742) CMV pp65 109-123

DRB1*15:01 NFPYLVAYQATVCARA (SEQ ID NO: 2743) HCV polyprotein

1582?1597, NS3

DRB1*15:01 GINAVAYYRGLDVSV (SEQ ID NO: 2744) HCV polyprotein

1411?1425, NS3

DRB1*15:01 TTVRLRAYMNTPGLPVC (SEQ ID NO: 2728) HCV NS3

1535-1551

DRB1*15:01 PVSKMRMATPLLMQA (SEQ ID NO: 2668) CLIP 87-101

DRB1*15:01 SSPKPWIYATSNLAS (SEQ ID NO: 2714) Rituximab Light

chain 41-55

DRB1*15:01 EWVAEIRSKSINSAT (SEQ ID NO: 2730) Infliximab Heavy

chain 46-60

DRB1*15:01 MHVSFVMAYPEMLAA (SEQ ID NO: 2745) Mtb PE family

protein

DRB1*15:01 MSQIMYNYPAMMAHA (SEQ ID NO: 2746) Mtb ESAT-6 like

protein

As further illustrative and non-limiting examples, the following antigens provided in Table 6 are known in the art to be relevant to celiac disease. In some embodiments, the TCR recognition domain can comprise one or more of the peptides provided in Table 6. In some embodiments, the TCR recognition domain can comprise a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of the peptides of Table 6. In some embodiments, the TCR recognition domain can comprise a sequences with at least 95% sequence identity to one of peptides of Table 6, wherein that sequence retains the wild-type activity of the peptide of Table 6.

TABLE 6
The sequences are, in the order presented, SEQ ID NOs:1000-2012.

ID	Type	Description	Toxicity	Form	HLADQ	Refs	SeqLen	Sequence

1	alpha-	alpha-gliadin	Toxic	Native	Unknown	41	22	VPVPQLQPQNPSQ
	gliadin	CT-1 (p1-p22						QQPQEQVPL
		of B 3142)
2	alpha-	alpha-gliadin	Toxic	Native	Unknown	41	31	VQQQQFPGQQQP
	gliadin	peptide CT-2						FPPQQPYPQPQPFP
		(p23-p53 of B						SQQPY
		3142)
3	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	34	19	VRVPVPQLQPQNP
	gliadin	p14 (p1-p19)						SQQQPQ
4	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	34	18	QNPSQQQPQEQVP
	gliadin	p15 (p11-p28)						LVQQQ
5	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	39, 34	20	FPGQQQPFPPQQP
	gliadin	p209						YPQPQPF
7	alpha-	alpha-gliadin	Immunogenic,	Native	HLA-DR	10	12	PQPQPFPSQQPY
	gliadin	(p44-p55)	Toxic
8	alpha-	Epitope DQ2-	Immunogenic	Native	DQ2	62	20	YLQLQPFPQPQLPY
	gliadin	alpha-I/II/III						PQPQLP
9	alpha-	alpha2-gliadin	Immunogenic	Native	DQ2, DQ8	17	15	YLQLQPFPQPQLPY
	gliadin	1420 (p56-						P
		p70)
10	alpha-	alpha2-gliadin	Immunogenic	Native	DQ2, DQ8	2, 25	33	LQLQPFPQPQLPYP
	gliadin	33-mer (p57-			(DQ2/8)			QPQLPYPQPQLPYP
		p89)						QPQPF
11	alpha-	Deamidated	Immunogenic	Deamidated	DQ2.5	61	33	LQLQPFPQPELPYP
	gliadin	alpha2-gliadin						QPELPYPQPELPYP
		33-mer (p57-						QPQPF
		p89)
12	alpha-	alpha-2 gliadin	Toxic,	Native	DQ2	51, 63	20	LQLQPFPQPQLPYP
	gliadin	G8 (p56-p75)	Immunogenic					QPQLPY
13	alpha-	alpha-2 gliadin	Immunogenic	Deamidated	DQ2	51	20	LQLQPFPQPELPYP
	gliadin	G9 (p56-p75;						QPQLPY
		E65)
14	alpha-	alpha-9 gliadin	Immunogenic	Native	DQ2	51	13	LQLQPFPQPQLPY
	gliadin	G5 (p56-p68)
15	alpha-	alpha-9 gliadin	Immunogenic	Deamidated	DQ2	51	13	LQLQPFPQPELPY
	gliadin	G5 (p56-p68;
		E65)
16	alpha-	Wheat	Immunogenic	Native	DQ2	62, 86	16	QLQPFPQPQLPYPQ
	gliadin	peptide W02						PQ
17	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	9	15	QLQPFPQPQLPYPQ
	gliadin	(p57-p71)						P
18	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	9	15	QLQPFPQPQLPYTH
	gliadin	(p57-p71; T69						P
		and H70)
19	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	9	15	QLRPFPQPQLPYPQ
	gliadin	(p57-p71; R59)						P
20	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	9	15	QLQPFPHPQLPYPH
	gliadin	(p57-p71; H63						P
		and H70)
21	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	9	15	QLQPFPQAQLPYPQ
	gliadin	(p57-p71;						P
		A63)
22	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	8	14	QLQPFPQPQLPYPQ
	gliadin	(p57-p70)
23	alpha-	Glia-alpha9	Immunogenic	Deamidated	DQ2	8	14	QLQPFPQPELPYPQ
	gliadin	(p57-p70; E65)
24	alpha-	alpha-9 gliadin	Immunogenic	Native	DQ2	14, 23,	12	QLQPFPQPQLPY
	gliadin	(p57-p68);				2, 84
		alpha2/alpha9
		gliadin
25	alpha-	alpha-9 gliadin	Immunogenic	Deamidated	DQ2	43, 14,	12	QLQPFPQPELPY
	gliadin	(p57-p68;				8, 84
		E65); alpha-l
26	alpha-	alpha-9 gliadin	Immunogenic	Native	DQ2	8	12	QLQPFPEPQLPY
	gliadin	epitope
		homolog (p57-
		p68; E63
		(considered
		native form of
		synthetic))
27	alpha-	alpha-9 gliadin	Immunogenic	Deamidated	DQ2	8	12	QLQPFPEPELPY
	gliadin	epitope
		homolog (p57-
		p68; E63 and
		E65 (tTG-
		treated form))
28	alpha-	alpha-9 gliadin	Immunogenic	Native	DQ2	8	12	QLQPFPQQQLPY
	gliadin	epitope
		homolog (p57-
		p68; Q64
		(considered
		native form of
		synthetic))
29	alpha-	alpha-9 gliadin	Immunogenic	Deamidated	DQ2	8	12	QLQPFPEQQLPY
	gliadin	epitope
		homolog (p57-
		p68; Q64 and
		E63 (tTG-
		treated form))
30	alpha-	alpha-9 gliadin	Immunogenic	Deamidated	DQ2	8	12	QLQPFPQQELPY
	gliadin	epitope
		homolog (p57-
		p68; Q64 and
		E65 (tTG-
		treated form))
31	alpha-	alpha-9 gliadin	Immunogenic	Deamidated	DQ2	8	12	QLQPFPEQELPY
	gliadin	epitope
		homolog (p57-
		p68; Q64, E63
		and E65 (tTG-
		treated form))
32	alpha-	DQ2-Glia-	Immunogenic	Native	DQ2	59	15	LQPFPQPQLPYPQP
	gliadin	alpha1						Q
		epitope (p58-
		p72)
33	alpha-	DQ2-Glia-	Immunogenic	Deamidated	DQ2	59	15	LQPFPQPELPYPQP
	gliadin	alpha1						Q
		epitope (p58-
		p72; E65)
34	alpha-	DQ2-Glia-	Immunogenic	Native	DQ2	59	15	LQPFPQSQLPYPQP
	gliadin	alpha1						Q
		epitope (p58-
		p72; S64)
35	alpha-	DQ2-Glia-	Immunogenic	Deamidated	DQ2	59	15	LQPFPQSELPYPQP
	gliadin	alpha1						Q
		epitope (p58-
		p72; S64 and
		E65)
36	alpha-	DQ2-alpha-I	Immunogenic	Native	DQ2	47	11	LQPFPQPQLPY
	gliadin	epitope (p58-
		p68)
37	alpha-	DQ2-alpha-l	Immunogenic	Deamidated	DQ2	47	11	LQPFPQPELPY
	gliadin	epitope (p58-
		p68; E65
		considered
		Deamidated
		form)
38	alpha-	Glia-alpha2	Immunogenic	Native	DQ2	8	17	QPFPQPQLPYPQP
	gliadin	(p60-p76)						QLPY
39	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	8	17	QPFPQPELPYPQPQ
	gliadin	(p60-p76; E66)						LPY
40	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	8	17	QPFPQPQLPYPQPE
	gliadin	(p60-p76; E73)						LPY
41	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	8	17	QPFPQPELPYPQPE
	gliadin	(p60-p76; E66						LPY
		and E73)
42	alpha-	Glia-alpha2	Immunogenic	Native	DQ2	45	10	QPFPQPQLPY
	gliadin	(p60-p69)
43	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	45	10	QPFPQPELPY
	gliadin	(p60-p69; E66)
44	alpha-	alpha2-gliadin	Immunogenic	Native	DQ2, DQ8	17	15	PFPQPQLPYPQPQL
	gliadin	1421 (p61-						P
		p75)
45	alpha-	Glia-alpha2	Immunogenic	Native	DQ2	59	11	PFPQPQLPYPQ
	gliadin	(p61-p71)
46	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	59	11	PFPQPELPYPQ
	gliadin	(p61-p71; E66)
47	alpha-	Glia-alpha2	Immunogenic	Native	DQ2	59	11	PFPQPQLPYTH
	gliadin	(p61-p71; T70
		and H71)
48	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	59	11	PFPQPELPYTH
	gliadin	(p61-p71; T70,
		H71 and E66)
49	alpha-	Glia-alpha2	Immunogenic	Native	DQ2	59	11	PFPHPQLPYPQ
	gliadin	(p61-p71;
		H64)
50	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	59	11	PFPHPELPYPQ
	gliadin	(p61-p71; H64
		and E66)
53	alpha-9	DQ2.5_glia_	Immunogenio	Native	DQ2.5	14, 90,	9	PFPQPQLPY
	gliadin	alpha-la				17, 23
54	alpha-9	DQ2.5_glia_	Immunogenic	Deamidated	DQ2.5	14, 90,	9	PFPQPELPY
	gliadin	alpha-1a				17, 23
55	alpha-	alpha-2 gliadin	Immunogenic	Native	DQ2	14, 1,	14	PQPQLPYPQPQLPY
	gliadin	1206 (p62-				2
		p75)
56	alpha-	alpha-II/alpha-	Immunogenic	Deamidated	DQ2	14	14	PQPQLPYPQPELPY
	gliadin	III epitope
		(p62-p75; E72)
57	alpha-	alpha-2 gliadin	Immunogenic	Deamidated	DQ2	14, 25	14	PQPELPYPQPQLPY
	gliadin	(p62-p75; E65)
58	alpha-	alpha-2 gliadin	Immunogenic	Deamidated	DQ2	14	14	PQPELPYPQPELPY
	gliadin	(p62-p75; E65
		and E72)
59	alpha-	G4-9A gliadin	Immunogenic	Deamidated	DQ2	51	14	PQPELPYPAPQLPY
	gliadin	(p62-p75; E65
		and A70)
60	alpha-	G4-11A gliadin	Immunogenic	Deamidated	DQ2	51	14	PQPELPYPQPALPY
	gliadin	(p62-p75; E65
		and A72)
61	alpha-	G4-12A gliadin	Immunogenic	Deamidated	DQ2	51	14	PQPELPYPQPQAPY
	gliadin	(p62-p75; E65
		and A73)
62	alpha-	G4-13A gliadin	Immunogenic	Deamidated	DQ2	51	14	PQPELPYPQPQLAY
	gliadin	(p62-p75; E65
		and A74)
63	alpha-	G4-14A gliadin	Immunogenic	Deamidated	DQ2	51	14	PQPELPYPQPQLPA
	gliadin	(p62-p75; E65
		and A70)
64	alpha-	alpha-2 gliadin	Immunogenic	Native	DQ2	25, 47	12	PQPQLPYPQPQL
	gliadin	(p62-p73)
65	alpha-	alpha-2 gliadin	Immunogenic	Deamidated	DQ2	25, 47	12	PQPELPYPQPQL
	gliadin	(p62-p73; E65)
66	alpha-	alpha-II (p62-	Immunogenic	Native	DQ2	43	11	PQPQLPYPQPQ
	gliadin	p72)
67	alpha-	alpha-II (p62-	Immunogenic	Deamidated	DQ2	43	11	PQPELPYPQPE
	gliadin	p72; E65 and
		E72)
68	alpha-2	CAUTION	Immunogenic	Native	DQ2.5	14, 17,	9	PQPQLPYPQ
	gliadin	100% match				23
		to one fungal
		protein and
		many wheat
		proteins with
		multiple
		epitopes
		DQ2.5_glia_
		alpha2
69	alpha-2	DQ2.5_glia	Immunogenic	Deamidated	DQ2.5	14, 17,	9	PQPELPYPQ
	gliadin	alpha2				23
70	alpha-	alpha2-gliadin	Immunogenic	Native	DQ2, DQ8	17	15	QPQLPYPQPQLPYP
	gliadin	1423 (p71-						Q
		p85)
71	alpha-	a-gliadin (p62-	Immunogenic	Native	DQ2	62	20	PQLPYPQPQLPYPQ
	gliadin	p84)						PQLPYP
72	alpha-	alpha-III-	Immunogenic	Native	DQ2	25	12	PQLPYPQPQLPY
	gliadin	gliadin (p62-
		p79)
73	alpha-	alpha-III-	Immunogenic	Deamidated	DQ2	25	12	PQLPYPQPELPY
	gliadin	gliadin (p62-
		p79; E76)
74	alpha-	alpha-I	Immunogenic	Native	DQ2	25	12	LQLPFPQPQLPY
	gliadin	epitope
75	alpha-	alpha-I	Immunogenic	Deamidated	DQ2	25	12	LQLPFPQPELPY
	gliadin	epitope
		Deamidated
		form
76	alpha-	Glia-alpha2	Immunogenic	Native	DQ2	57	25	PQLPQFLQPQPYPQ
	gliadin	25-mer (p64-						PQLPYPQPQPF
		p89)
77	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	57	25	PELPQFLQPQPYPQ
	gliadin	25-mer (p64-						PQLPYPQPQPF
		p89; E65)
78	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	57	25	PQLPQFLQPQPYPQ
	gliadin	25-mer (p64-						PELPYPQPQPF
		p89; E79)
79	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	57	25	PELPQFLQPQPYPQ
	gliadin	25-mer (p64-						PELPYPQPQPF
		p89; E65 and
		E79)
30	alpha-	alpha2-gliadin	Immunogenic	Native	DQ2	17	15	QLPYPQPQLPYPQP
	gliadin	1422 (p66-						Q
		p80)
81	alpha-	alpha-III	Immunogenic	Native	DQ2	47	13	QLPYPQPQLPYPQ
	gliadin	epitope (p66-
		p78)
82	alpha-	alpha-III	Immunogenic	Deamidated	DQ2	47	13	QLPYPQPELPYPQ
	gliadin	epitope (p66-
		p78; E72)
83	alpha-	Wheat	Immunogenic	Native	DQ2	62	12	LPYPQPQLPYPQ
	gliadin	peptide W01
84	alpha-3	DQ2.5_glia_	Immunogenic	Native	DQ2.5	23	9	PYPQPQLPY
	gliadin	alpha 1b
85	alpha-3	DQ2.5_glia_	Immunogenic	Deamidated	DQ2.5	23	9	PYPQPELPY
	gliadin	alpha 1b
86	alpha-	Glia-alpha2	Immunogenic	Native	DQ2	82	18	QPQPYPQPQLPYP
	gliadin	18-mer (p71-						QPQPF
		p89)
87	alpha-	Glia-alpha2	Immunogenic	Deamidated	DQ2	82, 57	18	QPQPYPQPELPYPQ
	gliadin	18-mer (p71-						PQPF
		p89; E79)
88	alpha-	Wheat	Immunogenic	Native	DQ2	62, 82	20	PQLPYPQPQLPYPQ
	gliadin	peptide W18						PQPFRP
89	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	20	PQLPYPQPELPYPQ
	gliadin	peptide W18						PQPFRP
90	alpha-	Wheat	Immunogenic	Native	DQ2	62	16	YPQPQLPYPQPQPF
	gliadin	peptide W18						RP
91	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	16	YPQPELPYPQPQPF
	gliadin	peptide W18						RP
92	alpha-	alpha2-gliadin	Immunogenic	Native	DQ2	17	15	YPQPQLPYPQPQPF
	gliadin	1424 (p76-						R
		p90)
93	alpha-20	DQ2.5_glia_	Immunogenic	Native	DQ2.5	90, 8	9	FRPQQPYPQ
	gliadin	alpha 3
94	alpha-20	DQ2.5_glia_	Immunogenic	Deamidated	DQ2.5	90, 8	9	FRPEQPYPQ
	gliadin	alpha 3
95	alpha-	Gliadin (p198-	Immunogenic	Native	DQ8	3	25	QQPQQQYPSGQGS
	gliadin	p222)			(DQ2/8,			FQPSQQNPQAQG
					DQ1/8)
96	alpha-	alpha-2 gliadin	Immunogenic	Native	DQ8	6	24	QQPQQQYPSGQGS
	gliadin	(p219-p242)						FQPSQQNPQAQ
		AJ133612
97	alpha-	alpha-2 gliadin	Immunogenic	Deamidated	DQ8	6	24	QQPQQQYPSGEGS
	gliadin	(p219-p242;						FQPSQENPQAQ
		E229 and
		E237)
		AJ133612
98	alpha-	alpha-2 gliadin	Immunogenic	Deamidated	DQ8	6	24	QQPQQQYPSGEGS
	gliadin	(p219-p242;						FQPSQQNPQAQ
		E229)
		AJ133612
99	alpha-	alpha-2 gliadin	Immunogenic	Deamidated	DQ8	6	24	QQPQQQYPSGQGS
	gliadin	(p219-p242;						FQPSQENPQAQ
		E237)
		AJ133612
100	alpha-	alpha-gliadin	Immunogenic	Native	DQ8	54	20	QPQQQYPSGQGSF
	gliadin	(p220-p239)						QPSQQNP
		P18573
101	alpha-	Gda09 (p202-	Immunogenic	Native	DQ8	3, 4	18	QQYPSGQGSFQPS
	gliadin	p219) P18573			(DQ2/8,			QQNPQ
					DQ1/8)
102	alpha-	Gda09 (p203-	Immunogenic	Native	DQ8	5	18	QYPSGQGSFQPSQ
	gliadin	p220) P18573						QNPQA
		(alpha-gliadin
		(alpha-I))
103	alpha-	Gda09 (p203-	Immunogenic	Deamidated	DQ8	5	18	QYPSGEGSFQPSQE
	gliadin	p220; E216)						NPQA
		P18573
		(alpha-gliadin
		(alpha-I)
104	alpha-	alpha2-gliadin	Immunogenic	Native	DQ8	17	15	YPSGQGSFQPSQQ
	gliadin	1447 (p226-						NP
		p240)
105	alpha-	Gliadin (p205-	Immunogenic	Native	DQ8	3	18	PSGQGSFQPSQQN
	gliadir	p222)			(DQ2/8)			PQAQG
106	alpha-	Gliadin (p205-	Immunogenic	Native	DQ8	46, 3	12	PSGQGSFQPSQQ
	gliadin	p216);			(DQ2/8)
		alpha2-gliadin
107	alpha-	Gliadin (p205-	Immunogenic	Native	DQ8	46, 3	11	PSGQGSFQPSQ
	gliadin	p215);			(DQ2/8)
		alpha2-gliadin
108	alpha-	Gda09 (p206-	Immunogenic	Native	DQ8	46, 4	12	SGQGSFQPSQQN
	gliadin	p217) P18573			(DQ2/8)
109	alpha-	Gda09 (p206-	Immunogenic	Deamidated	DQ8	46	12	SGQGSFQPSEQN
	gliadin	p217; E215)			(DQ2/8)
		P18573
110	alpha-	Gda09 (p206-	Immunogenic	Deamidated	DQ8	46, 83,	12	SGEGSFQPSQQN
	gliadin	p217; E208)			(DQ2/8)	4
		P18573
111	alpha-	Gda09 (p206-	Immunogenic	Deamidated	DQ8	46, 4	12	SGQGSFQPSQEN
	gliadin	p217; E216)			(DQ2/8)
		P18573
112	alpha-	Gda09 (p206-	Immunogenic	Deamidated	DQ8	3, 4	12	SGEGSFQPSQEN
	gliadin	p217; E208			(DQ2/8)
		and E216)
		P18573
113	alpha-	alpha2-gliadin	Immunogenic	Native	DQ8	6	9	GQGSFQPSQ
	gliadin	(p228-p236)			(DQ2/8)
115	alpha-2	CAUTION	Immunogenic	Native	DQ8	6, 90,	9	QGSFQPSQQ
	gliadin	100% identity			(DQ2/8)	3
		match to Dicot
		plant protein
		and many
		wheat family
		DQ8_glia_
		alpha 1
		DQ8.5_glia_
		alpha 1
116	alpha-2	DQ8_glia_	Immunogenic	Deamidated	DQ8	90, 17,	9	EGSFQPSQQ
	gliadin	alpha 1			(DQ2/8)	83
		DQ8.5_glia_
		alpha 1
117	alpha-2	DQ8_glia_	Immunogenic	Deamidated	DQ8	90, 17	9	QGSFQPSQE
	gliadin	alpha 1			(DQ2/8)
		DQ8.5_glia_
		alpha 1
118	alpha-2	DQ8_glia_	Immunogenic	Deamidated	DQ8	90, 17	9	EGSFQPSQE
	gliadin	alpha 1			(DQ2/8)
		DQ8.5_glia_
		alpha 1
119	alpha-	alpha2-gliadin	Immunogenic	Native	DQ8	17	15	GSFQPSQQNPQAQ
	gliadin	1448 (p231-						GS
		p245)
120	alpha-	alpha2-gliadin	Immunogenic	Native	DQ8 and	17	15	QAQGSVQPQQLPQ
	gliadin	1450 (p241-			weak DQ2			FE
		p255)
121	alpha-	Wheat	Immunogenic	Native	DQ2	62	20	MQLQPFPQPQLPY
	gliadin	peptide W02						PQPQLPY
122	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	20	MQLQPFPQPELPYP
	gliadin	peptide W02						QPQLPY
123	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	16	QLQPFPQPELPYPQ
	gliadin	peptide W02						PQ
124	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	16	ELQPFPQPELPYPQ
	gliadin	peptide W02						PQ
125	alpha-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQPQLPYPQ
	gliadin	peptide W02
126	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	12	QPFPQPELPYPQ
	gliadin	peptide W02
127	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	PQPFPPQLPYPQPQ
	gliadin	peptide W01						LPYPQP
128	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	20	PQPFPPQLPYPQPE
	gliadin	peptide W01						LPYPQP
129	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	12	LPYPQPELPYPQ
	gliadin	peptide W01
130	alpha-	Wheat	Immunogenic	Native	DQ2	62	20	VAHAIIMHQQQQQ
	gliadin	peptide W34						QQEQKQQ
131	alpha-	Wheat	Immunogenic	Native	DQ2	62	16	VAHAIIMHQQQQQ
	gliadin	peptide W34						QQE
132	alpha-	Analog of	Toxic	Native	DQ2	50	19	AGQQQPFPPQQPY
	gliadin	alpha-gliadin						PQPQPF
		(p31-p49;
		A31)
133	alpha-	Analog of	Toxic	Native	DQ2	50	19	LGQQQAFPPQQPY
	gliadin	alpha-gliadin						PQPQPF
		(p31-p49;
		A36)
134	alpha-	alpha20-	Immunogenic	Native	DQ2	8	16	PQPFRPQQPYPQP
	gliadin	gliadin (p91-						QPQ
		p106)
135	alpha-	alpha20-	Immunogenic	Deamidated	DQ2	8	16	PQPFRPEQPYPQPQ
	gliadin	gliadin (p93-						PQ
		106; E97)
136	alpha-	Glia-alpha20-	Immunogenic	Native	DQ2	19	14	PFRPQQPYPQPQP
	gliadin	gliadin (p93-						Q
		p106)
137	alpha-	Glia-alpha20-	Immunogenic	Deamidated	DQ2	19	14	PFRPEQPYPQPQPQ
	gliadin	gliadin (p93-
		p106; E97)
138	alpha-	Glia-alpha20-	Immunogenic	Native	DQ2	19	11	PQQPYPQPQPQ
	gliadin	gliadin (p96-
		106) minimal
		epitope
139	alpha-	Glia-alpha20-	Immunogenic	Deamidated	DQ2	19	11	PEQPYPQPQPQ
	gliadin	gliadin (p96-
		106; E97)
		minimal
		epitope,
		synthetic
140	alpha-	alpha-	Immunogenic	Native	DQ8	64, 30	10	QLIPCMDVVL
	gliadin	gliadin(p123-
		p132)
141	alpha-	alpha-gliadin	Toxic	Native	DQ2 (A1	35	12	LGQGSFRPSQQN
	gliadin	(p206-p217)			B8 DR3
					DQ2 and A
					24 B8 DR3
					13 DQ2)
142	alpha-	Peptide XT (1-	Toxic	Native	Unknown	29	55	VRVPVPQLQPQNP
	gliadin	55)						SQQQPQEQVPLVQ
								QQQFLGQQQPFPP
								QQPYPQPQPFPSQ
								QPY
143	alpha-	Peptide XT	Toxic	Native	Unknown	29	30	VRVPVPQLQPQNP
	gliadin	(p1-p30)						SQQQPQEQVPLVQ
								QQQF
144	alpha-	alpha-gliadin B	Immunogenic,	Native	Unknown	40	53	VPVPQLQPQNPSQ
	gliadin	3142 (p3-p55)	Toxic					QQPQEQVPLVQQQ
								QFGGQQQPFPPQQ
								PYPQPQPFPSQQPY
146	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	34	20	QVPLVQQQQFLGQ
	gliadin	p19 (p21-p40)						QQPFPPQ
147	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	39	19	QFLGQQQPFPPQQ
	gliadin	p134			(alpha1*			PYPQPQ
					0501,
					ß1*0201)
148	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	39	18	FLGQQQPFPPQQP
	gliadin	p135			(alpha1*			YPQPQ
					0501,
					ß1*0201)
149	alpha-	Peptide XT	Immunogenic,	Native	Unknown	10, 29	25	LGQQQPFPPQQPY
	gliadin	(p31-p55)	Toxic					PQPQPFPSQQPY
150	alpha-	alpha-gliadin	Immunogenic,	Native	DQ2	39, 11,	19	LGQQQPFPPQQPY
	gliadin	(p31-p49)	Toxic		(alpha1*	49, 34		PQPQPF
					0501,
					ß1*0201)
151	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	10, 39,	17	LGQQQPFPPQQPY
	gliadin	p126			(alpha1*	13, 15,		PQPQ
					0501,	66, 79
					ß1*0201)
152	alpha-	alpha-gliadin	Immunogenic,	Native	HLA-DR	10, 13,	13	LGQQQPFPPQQPY
	gliadin	(p31-p43)	Toxic			15, 72,
						79
153	alpha-	alpha-gliadin	Immunogenic	Native	DQ8	54	20	AMCNVYIPPYCAM
	gliadin	CAB76960						APFGIFG
		(p253-p272)
154	alpha-	alpha-gliadin	Immunogenic	Native	Unknown	42	16	CPQPFPSQQPYLQL
	gliadin	(proline-rich						QG
		domain)
155	alpha-	alpha-gliadin	Immunogenic	Native	Unknown	42	18	CPQLQPQNPSQQQ
	gliadin	(p5-p22)						PQEQG
		(proline-rich
		domain)
156	alpha-	alpha-gliadin	Toxic	Native	DQ2	37	20	SQQPYLQLQPFPQP
	gliadin	(p51-p70)						QLPYSQ
157	alpha-	Wheat	Immunogenic	Native	DQ2	62	20	LQLQPFPQPQLPYS
	gliadin	peptide W08						QPQPFR
158	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	20	LQLQPFPQPELPYS
	gliadin	peptide W08						QPQPFR
159	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	9	15	QLQPFPQPQLPYSQ
	gliadin	(p57-p71; S69)						P
160	alpha-	DQ2-Glia-	Immunogenic	Native	DQ2	59	15	LQPFPQPQLPYSQP
	gliadin	alpha1						Q
		epitope (p58-
		p72; S69)
161	alpha-	DQ2-Glia-	Immunogenic	Deamidated	DQ2	59	15	LQPFPQPELPYSQP
	gliadin	alpha1						Q
		epitope (p58-
		p72; S69 and
		E65)
162	alpha-	DQ2-Glia-	Immunogenic	Native	DQ2	59	15	LQPFPQSQLPYSQP
	gliadin	alpha1						Q
		epitope (p58-
		p72; S64 and
		S69)
163	alpha-	DQ2-Glia-	Immunogenic	Deamidated	DQ2	59	15	LQPFPQSELPYSQP
	gliadin	alpha1						Q
		epitope (p58-
		p72; S64, S69
		and E65)
164	alpha-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQPQLPYSQ
	gliadin	peptide W08
165	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	12	QPFPQPELPYSQ
	gliadin	peptide W08
166	alpha-	Glia-alpha	Immunogenic	Native	DQ2	59	11	PFPQPQLPYSQ
	gliadin
167	alpha-	Glia-alpha in	Immunogenic	Deamidated	DQ2	59	11	PFPQPELPYSQ
	gliadin	Deamidated
		form
168	alpha-	alpha-gliadin	Toxic	Native	DQ2	11	19	QQYPLGQGSFRPS
	gliadin	(p202-p220)						QQNPQA
169	alpha-	alpha-gliadin	Immunogenic	Native	DQ8	54	20	VYIPPYCTIAPFG
	gliadin	CAB76961						IFGTNYR
		(p251-p270)
170	alpha-	Wheat	Immunogenic	Native	DQ2	62	20	LQLQPFPQPQLPYL
	gliadin	peptide W13						QPQPFR
171	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	20	LQLQPFPQPELPYL
	gliadin	peptide W13						QPQPFR
172	alpha-	Glia-alpha9	Immunogenic	Native	DQ2	9	15	QLQPFPQPQLPYLQ
	gliadin	(p57-p71; L69)						P
173	alpha-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQPQLPYLQ
	gliadin	peptide W13
174	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	12	QPFPQPELPYLQ
	gliadin	peptide W13
175	alpha-	alpha-gliadin	Immunogenic	Native	Unknown	60	17	PPYCTIVPFGIF
	gliadin	4037						GTNYR
176	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	62	20	LQLQPFPQPQLPYP
	gliadin							QPQPFR
177	alpha-	alpha-Glia	Immunogenic	Native	DQ2	57	17	QLQPFPQPQLPYPQ
	gliadin	(p57-p73)						PQP
178	alpha-	alpha-Glia	Immunogenic	Deamidated	DQ2	57	17	QLQPFPQPELPYPQ
	gliadin	(p57-p73;						PQP
		E65)
179	alpha-	alpha-Glia	Immunogenic	Native	DQ2	57	17	QLQPFPQPTLPYPQ
	gliadin	(p57-p73; T65						PQS
		and S73)
180	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	28	17	QLQPFPQPQLPYPQ
	gliadin	(p57-p73; S73)						PQS
181	alpha-	alpha-gliadin	Immunogenic	Deamidated	DQ2	28	17	QLQPFPQPELPYPQ
	gliadin	(p57-p73; S73						PQS
		and E65)
182	alpha-	Wheat	Immunogenic	Native	DQ2	62	20	LQPFPQPQPFLPQL
	gliadin	peptide W09						PYPQPQ
183	alpha-	alpha-Glia	Immunogenic	Native	DQ2	57	17	PQPQPFLPQLPYPQ
	gliadin	AG11 (p78-						PQS
		p95)
184	alpha-	alpha-Glia	Immunogenic	Deamidated	DQ2	57	17	PQPQPFLPELPYPQ
	gliadin	AG11 (p78-						PQS
		p95; E86)
185	alpha-	Wheat	Immunogenic	Native	DQ2	62	14	QPQPFLPQLPYPQP
	gliadin	peptide W09
186	alpha-	Wheat	Immunogenic	Deamidated	DQ2	62	14	EPQPFLPELPYPQP
	gliadin	peptide W09
187	alpha-	Wheat	Immunogenic	Native	DQ2	62	12	PQPFLPQLPYPQ
	gliadin	peptide W09
188	alpha-	alpha-gliadin	Immunogenic	Native	DQ2	34	20	FPGQQQQFPPQQP
	gliadin	p211						YPQPQPF
189	alpha-	alpha-Glia	Immunogenic	Native	DQ2	57	17	PQPQPFPPQLPYPQ
	gliadin	AG12 (p82-						PQS
		p98
190	alpha-	alpha-Glia	Immunogenic	Deamidated	DQ2	57	17	PQPQPFPPELPYPQ
	gliadin	AG12 (p82-						PQS
		p98; E90)
191	omega-	Gliadin	Immunogenic	Native	DQ8	54	20	PFTQPQQPTPIQPQ
	gliadin	AAG17702						QPFPQQ
		(p80-p99)
192	omega-	Wheat	Immunogenic	Native	DQ2	62	11	PFTQPQQPTPI
	gliadin	peptide W27
193	omega-	Gliadin	Immunogenic	Native	DQ8	54	20	TPIQPQQPFPQQP
	gliadin	AAG17702						QQPQQPF
		(p88-p107)
194	omega-	Wheat	Immunogenic	Native	DQ2	62	11	TPIQPQQPFPQ
	gliadin	peptide W25
195	omega-	Wheat	Immunogenic	Native	DQ2	62	12	PQQPFPQQPQQP
	gliadin	peptide W30;
		Rye peptide
		R28
197	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	20	PQQPQQPQQPFPQ
	gliadin							PQQPFPW
198	omega-	Epitope DQ2-	Immunogenic	Native	DQ2	62	20	PQQPQQPFPQPQQ
	gliadin	omega-I/II						PFPWQPQ
199	omega-	p4-p18	Immunogenic	Native	DQ2	62	15	PQQPQQPFPQPQQ
	gliadin	omega-gliadin						PF
		of AAG17702
		(p81-p102)
200	omega-	p5-p19	Immunogenic	Native	DQ2	62	15	QQPQQPFPQPQQP
	gliadin	omega-gliadin						FP
		of AAG17702
		(p81-p102)
201	omega-	DQ2-omega-1	Immunogenic	Native	DQ2	57	17	QPQQPFPQPQQPF
	gliadin	omega-Glia						PWQP
		(p102-p118)
202	omega-	DQ2-omega-1	Immunogenic	Deamidated	DQ2	57	17	QPEQPFPQPQQPF
	gliadin	omega-Glia						PWQP
		(p102-p118;
		E104)
203	omega-	Wheat	Immunogenic	Deamidated	DQ2	62, 86	17	QPEQPFPQPEQPFP
	gliadin	peptide W03,						WQP
		W19, B01
204	omega-	omega-Glia	Immunogenic	Deamidated	DQ2.5	61	17	QPQQPFPQPEQPF
	gliadin	17mer						PWQP
205	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	14	KPFPQPEQPFPWQ
	gliadin	AAG17702						P
		substituted by
		Lysine (p89-
		p102; E95
		K89)
206	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	14	QKFPQPEQPFPWQ
	gliadin	AAG17702						P
		substituted by
		Lysine (p89-
		p102; E95
		K90)
207	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	14	QPKPQPEQPFPWQ
	gliadin	AAG17702						P
		substituted by
		Lysine (p89-
		p102; E95
		K91)
208	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	14	QPFPQPEQPFKWQ
	gliadin	AAG17702						P
		substituted by
		Lysine (p89-
		p102; E95
		K99
209	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	14	QPFPQPEQPFPKQP
	gliadin	AAG17702
		substituted by
		Lysine (p89-
		p102; E95
		K100)
210	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	14	QPFPQPEQPFPWK
	gliadin	AAG17702						P
		substituted by
		Lysine (p89-
		p102; E95
		K101)
211	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	14	QPFPQPEQPFPWQ
	gliadin	AAG17702						K
		substituted by
		Lysine (p89-
		p102; E95
		K102)
212	omega-	p6-p20	Immunogenic	Native	DQ2	62	15	QPQQPFPQPQQPF
	gliadin	omega-gliadin						PW
		of AAG17702
		(p81-p102)
213	omega-	p7-p21	Immunogenic	Native	DQ2	62	15	PQQPFPQPQQPFP
	gliadin	omega-gliadin						WQ
		of AAG17702
		(p81-p102)
214	omega-	p8-p22	Immunogenic	Native	DQ2	62	15	QPFPQPQQPFPW
	gliadin	omega-gliadin						QP
		of AAG17702
		(p81-p102),
		Wheat
		peptide W3
215	omega-	Wheat	Immunogenic	Deamidated	DQ2	62	15	EQPFPQPEQPFPW
	gliadin	peptide W03						QP
216	omega-	Wheat	Immunogenic	Native	DQ2	62	20	QPFPQPQQPFPWQ
	gliadin	peptide W03,						PQQPFPQ
		W19, Barley
		peptide B01
217	omega-	Wheat	Immunogenic	Deamidated	DQ2	62	20	QPFPQPEQPFPWQ
	gliadin	peptide W03,						PQQPFPQ
		W19, Barley
		peptide B01
218	omega-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQPQQPFPW
	gliadin	peptide W03,
		Barley peptide
		B01
219	omega-	Wheat	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPFPW
	gliadin	peptide W03,
		Barley peptide
		B01
220	omega-II	DQ2.5_glia_	Immunogenic	Deamidated	DQ2.5	88, 90,	9	PQPEQPFPW
	gliadin	omega 2				62
221	omega-II	DQ2.5_glia_	Immunogenic	Native	DQ2	88, 90,	9	PQPQQPFPW
	gliadin	omega 2				62
222	omega-	Wheat	Immunogenic	Native	DQ2	62	12	PFPWQPQQPFPQ
	gliadin	peptide W19,
		Barley peptide
		B19
223	omega-	Wheat	Immunogenic	Deamidated	DQ2	62	12	PFPWQPEQPFPQ
	gliadin	peptide W19
224	omega-	Wheat	Immunogenic	Native	DQ2	62	20	PLQPQQPFPQQPQ
	gliadin	peptide W30						QPFPQPQ
225	omega-	omega-gliadin	Immunogenic	Native	DQ2	62	20	FPQQPQQPFPQPQ
	gliadin							LPFPQQS
226	omega-	Wheat	Immunogenic	Native	DQ2	62	20	QQPQQPFPQPQLP
	gliadin	peptide W06						FPQQSEQ
227	omega-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQPQLPFPQ
	gliadin	peptide W06
228	omega-	Gliadin	Immunogenic	Native	DQ8	54	20	QQPFPQQPQQPFP
	gliadin	AAG17702						QPQQPIP
		p173-p192
229	omega-	Wheat	Immunogenic	Native	DQ2	62	12	PFPQQPQQPFPQ
	gliadin	peptide W32,
		Barley peptide
		B25, Rye
		peptide R26
230	omega-	Wheat	Immunogenic	Native	DQ2	62	20	PQQPQQPFPQPQQ
	gliadin	peptide W04						PIPVQPQ
231	omega-	Wheat	Immunogenic	Native	DQ2	62	11	PFPQPQQPIPV
	gliadin	peptide W04
232	omega-	Gliadin	Immunogenic	Native	DQ8	54	20	QPQQPIPVQPQQS
	gliadin	AAG17702						FPQQSQQ
		p186-p205
233	omega-	Wheat	Immunogenic	Native	DQ2	62	20	FPELQQPIPQQPQQ
	gliadin	peptide W20						PFPLQP
234	omega-	Wheat	Immunogenic	Native	DQ2	62	12	PIPQQPQQPFPL
	gliadin	peptide W20
235	omega-	Gliadin	Immunogenic	Native	DQ8	54	20	PQQPQQPFPLQPQ
	gliadin	AAG17702						QPFPQQP
		p225-p244
236	omega-	Wheat	Immunogenic	Native	DQ2	62	12	PFPLQPQQPFPQ
	gliadin	peptide W26,
		Barley peptide
		B20
237	omega-	Gliadin	Immunogenic	Native	DQ8	54	20	PFPQQPQQPFPQQ
	gliadin	AAG17702						PQQSFPQ
		p239-p258
246	omega5-	Glu-5 peptide	Immunogenic	Native	DQ2	21	12	QQQQIPQQPQQF
	gliadin/	epitope in
	LMW	native form
	glutenin
247	omega5-	Glu-5 peptide	Immunogenic	Native	DQ2	21	12	QQQQLPQQPQQF
	gliadin/	epitope in
	LMW	native form
	glutenin
248	omega5-	Glu-5 peptide	Immunogenic	Deamidated	DQ2	21	12	QEQQIPEQPQQF
	gliadin/	epitope in
	LMW	Deamidated
	glutenin	form
249	omega5-	Glu-5 peptide	Immunogenic	Deamidated	DQ2	21	12	QEQQLPEQPQQF
	gliadin/	epitope in
	LMW	Deamidated
	glutenin	form
252	omega5-	CAUTION	Immunogenic	Native	DQ2	19	9	QIPQQPQQF
	gliadin/	100% matches
	LMW	with 4 fungal
	glutenin	proteins but
		multiple
		epitopes on
		Triticum Glu-5
		minimal
		epitope in
		native form
253	omega5-	Glu-5 minimal	Immunogenic	Deamidated	DQ2	19	9	QIPEQPQQF
	gliadin/	epitope in
	LMW	Deamidated
	glutenin	form
254	omega5-	CAUTION	Immunogenic	Native	DQ2	19	9	QLPQQPQQF
	gliadin/	100% match
	LMW	with fungal
	glutenin	and parasite
		proteins, less
		with Glu-5
		minimal
		epitope in
		native form
255	omega5-	Glu-5 minimal	Immunogenic	Deamidated	DQ2	19	9	QLPEQPQQF
	gliadin/	epitope in
	LMW	Deamidated
	glutenin	form
256	omega5-	Glu-5 minimal	Immunogenic	Deamidated	DQ2	44	9	EIPEQPQQF
	gliadin/	epitope in			(DQ2.2 and
	LMW	Deamidated			DQ2.5)
	glutenin	form
257	omega5-	Glu-5 minimal	Immunogenic	Deamidated	DQ2	44	9	ELPEQPQQF
	gliadin/	epitope in			(DQ2.2 and
	LMW	Deamidated			DQ2.5)
	glutenin	form
258	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQISQPQIPQQQQI
	gliadin							PQQPQQF
	or LMW
	glutenin
259	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
260	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
261	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQIPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
262	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQIPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
263	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQQQ
	gliadin	Deamidated						QIPQQPQQF
	or LMW	form
	glutenin
264	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQEQ
	gliadin	Deamidated						QIPQQPQQF
	or LMW	form
	glutenin
265	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQQQE
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
266	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQEQQ
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
267	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQQQE
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
268	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQIPQEQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
269	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQEQQ
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
270	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQQQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
271	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQEQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
272	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQEQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
273	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQEQE
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
274	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQISQPQLPQQQQI
	gliadin							PQQPQQF
	or LMW
	glutenin
275	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
276	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
277	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQLPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
278	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQLPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
279	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
280	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
281	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
282	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
283	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
284	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQLPQEQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
285	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQEQ
	gliadin	Deamidated						QIPQQPQQF
	or LMW	form
	glutenin
286	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQQQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
287	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQEQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
288	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQEQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
289	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQEQE
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
290	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQISQPQIPQQQQL
	gliadin							PQQPQQF
	or LMW
	glutenin
291	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
292	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
293	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQIPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
294	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQIPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
295	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
296	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
297	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
298	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
299	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
300	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQIPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
301	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQEQ
	gliadin	Deamidated						QLPQQPQQF
	or LMW	form
	glutenin
302	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQQQ
	gliadin	Deamidated						ELPQQPQQF
	or LMW	form
	glutenin
303	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQIPQEQ
	gliadin	Deamidated						ELPQQPQQF
	or LMW	form
	glutenin
304	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPEIPQEQ
	gliadin	Deamidated						ELPQQPQQF
	or LMW	form
	glutenin
305	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPEIPQEQ
	gliadin	Deamidated						ELPQQPQQF
	or LMW	form
	glutenin
306	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQLSQPQIPQQQQI
	gliadin							PQQPQQF
	or LMW
	glutenin
307	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQIPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
308	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
309	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQIPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
310	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQIPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
311	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
312	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQIPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
313	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQIPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
314	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
315	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
316	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQIPQEQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
317	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQEQQ
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
318	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQQQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
319	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQIPQEQE
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
320	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQEQ
	gliadin	Deamidated						EIPQQPQQF
	or LMW	form
	glutenin
321	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQEQE
	gliadin	Deamidated						IPQQPQQF
	or LMW	form
	glutenin
322	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQLSQPQLPQQQQ
	gliadin							IPQQPQQF
	or LMW
	glutenin
323	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
324	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
325	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQLPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
326	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQLPQQQE
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
327	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQQQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
328	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
329	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
330	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
331	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
332	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQLPQEQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
333	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQEQQI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
334	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQQQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
335	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQEQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
336	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQEQEI
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
337	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQE
	gliadin	Deamidated						QEIPQQPQQF
	or LMW	form
	glutenin
338	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQLSQPQIPQQQQ
	gliadin							LPQQPQQF
	or LMW
	glutenin
339	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2		21	QELSQPQIPQQQQL
	gliadin	Deamidated				19		PQQPQQF
	or LMW	form
	glutenin
340	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
341	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQIPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
342	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQIPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
343	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
344	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQIPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
345	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQIPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
346	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
347	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
348	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQIPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
349	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
350	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
351	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQIPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
352	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPEIPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
353	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPEIPQEQ
	gliadin	Deamidated						ELPQQPQQF
	or LMW	form
	glutenin
354	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQISQPQLPQQQQ
	gliadin							LPQQPQQF
	or LMW
	glutenin
355	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
356	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
357	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQLPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
358	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQLPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
359	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
360	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
361	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
362	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
363	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
364	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPQLPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
365	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
366	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
367	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPQLPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
368	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQISQPELPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
369	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QEISQPELPQEQ
	gliadin	Deamidated						ELPQQPQQF
	or LMW	form
	glutenin
370	gamma-	Glu-5	Immunogenic	Native	DQ2	19	21	QQLSQPQLPQQQQ
	gliadin							LPQQPQQF
	or LMW
	glutenin
371	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQQQQ
	gliadin	Deamidated						LPQQPQQF
	or LMW	form
	glutenin
372	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQQQQ
	gliadin	Deamidated						LPQQPQQF
	or LMW	form
	glutenin
373	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQLPQEQQ
	gliadin	Deamidated						LPQQPQQF
	or LMW	form
	glutenin
374	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQLPQQQE
	gliadin	Deamidated						LPQQPQQF
	or LMW	form
	glutenin
375	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQQQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
376	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
377	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
378	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
379	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
380	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPQLPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
381	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQEQQL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
382	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQQQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
383	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPQLPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
384	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QQLSQPELPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
385	gamma-	Glu-5 in	Immunogenic	Deamidated	DQ2	19	21	QELSQPELPQEQEL
	gliadin	Deamidated						PQQPQQF
	or LMW	form
	glutenin
386	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	54	20	QQFLQPQQPFPQQ
	gliadin	P08079						PQQPYPQ
387	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	54	17	QQFLQPQQPFPQQ
	gliadin	P08079						PQQP
388	gamma-	gamma-5	Immunogenic	Native	DQ2	48	26	FLQPQQPFPQQPQ
	gliadin	gliadin (p59-						QPYPQQPQQPFPQ
		p84)
389	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPQQPQ
	gliadin	gliadin (p59-						QPYPQQPQQPFPQ
		p84; E63)
390	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLOPQQPFPEQPQ
	gliadin	gliadin (p59-						QPYPQQPQQPFPQ
		p84; E68)
391	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPQQPE
	gliadin	gliadin (p59-						QPYPQQPQQPFPQ
		p84; E71)
392	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPQQPQ
	gliadin	gliadin (p59-						QPYPEQPQQPFPQ
		p84; E76)
393	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPQQPQ
	gliadin	gliadin (p59-						QPYPQQPEQPFPQ
		p84; E79)
394	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPEQPQQ
	gliadin	gliadin (p59-						PYPQQPQQPFPQ
		p84; E63 and
		E68)
395	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPQQPEQ
	gliadin	gliadin (p59-						PYPQQPQQPFPQ
		p84; E63 and
		E71)
396	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPQQPQ
	gliadin	gliadin (p59-						QPYPEQPQQPFPQ
		p84; E63 and
		E76)
397	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPQQPQ
	gliadin	gliadin (p59-						QPYPQQPEQPFPQ
		p84; E63 and
		E79)
398	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLOPQQPFPEQPEQ
	gliadin	gliadin (p59-						PYPQQPQQPFPQ
		p84; E68 and
		E71)
399	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLOPQQPFPEQPQ
	gliadin	gliadin (p59-						QPYPEQPQQPFPQ
		p84; E68 and
		E76)
400	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPEQPQ
	gliadin	gliadin (p59-						QPYPQQPEQPFPQ
		p84; E68 and
		E79)
401	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPQQPE
	gliadin	gliadin (p59-						QPYPEQPQQPFPQ
		p84; E71 and
		E76)
402	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPQQPE
	gliadin	gliadin (p59-						QPYPQQPEQPFPQ
		p84; E71 and
		E79)
403	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPQQPQ
	gliadin	gliadin (p59-						QPYPEQPEQPFPQ
		p84; E76 and
		E79)
404	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPEQPEQ
	gliadin	gliadin (p59-						PYPQQPQQPFPQ
		p84; E63, E68
		and E71)
405	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPEQPQQ
	gliadin	gliadin (p59-						PYPEQPQQPFPQ
		p84; E63, E68
		and E76)
406	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPEQPQQ
	gliadin	gliadin (p59-						PYPQQPEQPFPQ
		p84; E63, E68
		and E79)
407	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPEQPEQ
	gliadin	gliadin (p59-						PYPEQPQQPFPQ
		p84; E68, E71
		and E76)
408	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPEQPEQ
	gliadin	gliadin (p59-						PYPQQPEQPFPQ
		p84; E68, E71
		and E79)
409	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPQQPFPQQPE
	gliadin	gliadin (p59-						QPYPEQPEQPFPQ
		p84; E71, E76
		and E79)
410	gamma-	gamma-5	Immunogenic	Deamidated	DQ2	48	26	FLQPEQPFPEQPEQ
	gliadin	gliadin (p59-						PYPEQPEQPFPQ
		p84; E63, E68,
		E71,E76 and
		E79)
411	gamma-	gamma-5	Immunogenic	Native	DQ2	57, 23,	20	LQPQQPFPQQPQQ
	gliadin	gliadin (p60-				48, 2		PYPQQPQ
		p79); DQ2-
		gamma-V
		gamma-Glia
		(p78-p97);
		gamma-3 and
		gamma-5
		peptide 1317
412	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPEQPFPQQPQQ
	gliadin	gamma-Glia				2		PYPQQPQ
		(p78-p97;
		E81)
413	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPQQPFPEQPQQ
	gliadin	gamma-Glia				2		PYPQQPQ
		(p78-p97;
		E86)
414	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPQQPFPQQPEQ
	gliadin	gamma-Glia				2		PYPQQPQ
		(p78-p97;
		E89)
415	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPQQPFPQQPQQ
	gliadin	gamma-Glia				2		PYPEQPQ
		(p78-p97;
		E94)
416	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPEQPFPEQPQQP
	gliadin	gamma-Glia				2		YPQQPQ
		(p78-p97;
		E81 and E86)
417	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPEQPFPQQPEQP
	gliadin	gamma-Glia				2		YPQQPQ
		(p78-p97;
		E81 and E89)
418	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPEQPFPQQPQQ
	gliadin	gamma-Glia				2		PYPEQPQ
		(p78-p97;
		E81 and E94)
419	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPQQPFPEQPEQP
	gliadin	gamma-Glia				2		YPQQPQ
		(p78-p97;
		E86 and E89)

420	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPQQPFPEQPQQ
	gliadin	gamma-Glia				2		PYPEQPQ
		(p78-p97;
		E86 and E64)
421	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPQQPFPQQPEQ
	gliadin	gamma-Glia				2		PYPEQPQ
		(p78-p97;
		E89 and E94)
422	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPEQPFPEQPEQP
	gliadin	gamma-Glia				2		YPQQPQ
		(p78-p97;
		E81, E86 and
		E89)
423	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPEQPFPEQPQQP
	gliadin	gamma-Glia				2		YPEQPQ
		(p78-p97;
		E81, E86 and
		E94)
424	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPQQPFPEQPEQP
	gliadin	gamma-Glia				2		YPEQPQ
		(p78-p97;
		E86, E89 and
		E94)
425	gamma-	DQ2-gamma-V	Immunogenic	Deamidated	DQ2	57, 23,	20	LQPEQPFPEQPEQP
	gliadin	gamma-Glia				2		YPEQPQ
		(p78-p97;
		E81, E86, E89
		and E94)
426	gamma-	gamma3/gam	Immunogenic	Native	DQ2	25	17	PQQPFPQQPQQPY
	gliadin	ma4						PQQP
427	gamma-	gamma5 (p62-	Immunogenic	Native	DQ2	25	13	PQQPFPQQPQQPY
	gliadin	p74)
428	gamma-	gamma5 (p62-	Immunogenic	Deamidated	DQ2	25	13	PQQPFPEQPQQPY
	gliadin	p74; E68)
429	gamma-	gamma5 (p62-	Immunogenic	Deamidated	DQ2	25	13	PEQPFPEQPQQPY
	gliadin	p74; E63 and
		E68)
430	gamma-	gamma5 (p62-	Immunogenic	Deamidated	DQ2	25	13	PQQPFPEQPEQPY
	gliadin	p74; E68 and
		E71)
431	gamma-	gamma5 (p62-	Immunogenic	Deamidated	DQ2	25	13	PEQPFPEQPEQPY
	gliadin	p74; E63, E68
		and E71)
432	gamma-	gamma-5	Immunogenic	Native	DQ2	47, 48	11	PQQPFPQQPQQ
	gliadin	gliadin (p62-
		p72)
433	gamma-	gamma5-	Immunogenic	Deamidated	DQ2	47	11	PQQPFPEQPQQ
	gliadin	gliadin (p62-
		p72; E68)
434	gamma-	gamma5 (p62-	Immunogenic	Deamidated	DQ2	25	11	PEQPFPEQPEQ
	gliadin	p72; E68, E63
		and E71)
435	gamma-	gamma23mer	Immunogenic	Native	DQ2.5	61	23	QQPFPQQPQQPYP
	gliadin							QQPQQPFPQP
436	gamma-	gamma23mer	Immunogenic	Deamidated	DQ2.5	61	23	EQPFPEQPEQPYPE
	gliadin	(in considered						QPEQPFPQP
		Deamidated
		form)
437	gamma-	gamma5-	Immunogenic	Native	DQ2	21	14	QQPFPQQPQQPYP
	gliadin	gliadin (p60-						Q
		p79)
438	gamma-5	DQ2.5_glia_ga	Immunogenic	Native	DQ2.5	25, 90,	9	QQPFPQQPQ
	gliadin	mma 5				23
439	gamma-5	DQ2.5_glia_ga	Immunogenic	Deamidated	DQ2.5	25, 90,	9	QQPFPEQPQ
	gliadin	mma 5				23
440	gamma-	CAUTION	Immunogenic	Deamidated	DQ2	25	9	EQPFPEQPE
	gliadin	100% match
		to Archaea
		protein lower
		to others and
		to gamma5
		(p63-p71; E63,
		E68 and E71)
441	gamma-	DQ2-gamma-	Immunogenic	Native	DQ2	57, 48,	15	PFPQQPQQPYPQQ
	gliadin	III gamma-Glia				23, 2		PQ
		(p83-p97)
442	gamma-	DQ2-gamma-	Immunogenic	Deamidated	DQ2	57, 48,	15	PFPEQPQQPYPQQ
	gliadin	III gamma-Glia				23, 2		PQ
		(p83-p97;
		E86)
443	gamma-	DQ2-gamma-	Immunogenic	Deamidated	DQ2	57, 48,	15	PFPQQPEQPYPQQ
	gliadin	III gamma-Glia				23, 2		PQ
		(p83-p97;
		E89)
444	gamma-	DQ2-gamma-	Immunogenic	Deamidated	DQ2	57, 48,	15	PFPEQPEQPYPQQP
	gliadin	III gamma-Glia				23, 2		Q
		(p83-p97; E86
		and E89)
445	gamma-	Wheat	Immunogenic	Native	DQ2	62	12	PFPQQPQQPYPQ
	gliadin	peptide W23
446	gamma-	gamma-gliadin	Immunogenic	Native	DQ8, DQ2	17	15	FPQQPQQPYPQQP
	gliadin	(p66-p80)						QQ
		AJ416339
447	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ8	17, 83	15	FPEQPQQPYPQQP
	gliadin	(p66-p80; E68)
		AJ416339						QQ
448	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	17	15	FPQQPEQPYPQQP
	gliadin	(p66-p80; E71)						QQ
		AJ416339
449	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ8, DQ2	17	15	FPQQPQQPYPEQP
	gliadin	(p66-p80; E76)						QQ
		AJ416339
450	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ8	17	15	FPEQPQQPYPEQP
	gliadin	(p66-p80; E71						QQ
		and 76)
		AJ416339
451	gamma-	gamma5 (p66-	Immunogenic	Native	DQ2	23	13	FPQQPQQPYPQQP
	gliadin	p78)
452	gamma-	gamma5 (p66-	Immunogenic	Deamidated	DQ2	23	13	FPEQPEQPYPQQP
	gliadin	p78; E68 and
		E71)
453	gamma-	gamma5 (p66-	Immunogenic	Deamidated	DQ2	23	13	FPEQPQEPYPQQP
	gliadin	p78; E68 and
		E72)
454	gamma-	gamma5 (p66-	Immunogenic	Native	DQ2	25	12	FPQQPQQPYPQQ
	gliadin	p77)
455	gamma-	gamma5 (p66-	Immunogenic	Deamidated	DQ2	25	12	FPQQPEQPYPQQ
	gliadin	p77; E71)
456	gamma-	gamma5 (p66-	Immunogenic	Deamidated	DQ2	25	12	FPEQPEQPYPEQ
	gliadin	p77; E68, E71
		and E76)
457	gamma-	gamma5 (p67-	Immunogenic	Deamidated	DQ2	25	11	PEQPEQPYPEQ
	gliadin	p77; E68, E71
		and E76)
458	gamma-1	CAUTION	Immunogenic	Native	DQ2.5/	17, 23,	9	QPQQPYPQ
	and	100% matches			DQ8	78
	gamma 5	to many
	gliadin	fungal and
		parasite
		proteins as
		well as
		DQ2.5_glia_
		gamma 3
		DQ8_glia_
		gamma 1b
459	gamma 5	DQ2.5_glia_	Immunogenic	Deamidated	DQ2.5/	25, 17,	0	EQPEQPYPE
	gliadin	gamma 3			DQ8	78
		DQ8_glia_
		gamma 1b
460	gamma-1	DQ2.5_glia_	Immunogenic	Deamidated	DQ2.5/	25, 17,	9	EQPQQPYPE
	gliadin	gamma 3			DQ8	7
		DQ8_glia_
		gamma 1b
461	gamma-1	DQ2.5_glia_	Immunogenic	Deamidated	DQ2.5/	17, 78	9	EQPQQPFPE
	gliadin	gamma 3			DQ8
		DQ8_glia_
		gamma 1b
462	gamma-III	CAUTION	Immunogenic	Deamidated	DQ2.5/	17, 78	9	QQPEQPYPQ
	gliadin	100% identity			DQ8
		matches to
		fungal and
		lower to other
		fungal
		proteins as
		DQ2.5_glia_
		gamma 3
		DQ8_glia_
		gamma 1b
463	gamma-	Predicted	Immunogenic	Native	DQ8	22	14	QQPYPQQPQQPFP
	gliadin	gamma-gliadin			(DQ2/8)			Q
		peptide
464	gamma-	CAUTION	Immunogenic	Native	DQ2	25	9	QQPYPQQPQ
	gliadin	many 100%
		identity
		matches to
		legume and
		bacterial
		proteins and
		also gamma-
		Vib gliadin
465	gamma-	CAUTION	Immunogenic	Deamidated	DQ2	25	9	EQPYPQQPQ
	gliadin	100% match
		to bacterial
		protein and
		lower to
		others and
		gamma-Vib
		gliadin in
		Deamidated
		form
466	gamma-	CAUTION	Immunogenic	Deamidated	DQ2	25	9	QQPYPEQPQ
	gliadin	100% identity
		to bacterial
		protein and
		lower to
		others and
		gamma-Vib
		gliadin in
		Deamidated
		form
467	gamma-	gamma-Vib	Immunogenic	Deamidated	DQ2	25	9	EQPYPEQPQ
	gliadin	gliadin in
		Deamidated
		form
468	gamma-	CAUTION	Immunogenic	Native	DQ2	44	9	PYPQQPQQP
	gliadin	many 100%			(DQ2.2 and
		matches to			DQ2.5)
		fish, fungi and
		other non-
		wheat family
		proteins also a
		few to Secalin
		Glia-gamma2
469	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	44, 83	9	PYPEQPQQP
	gliadin	in Deamidated			(DQ2.2 and
		form			DQ2.5)
470	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	44	9	PYPQQPEQP
	gliadin	in Deamidated			(DQ2.2 and
		form			DQ2.5)
471	gamma-	CAUTION	Immunogenic	Deamidated	DQ2	44	9	PYPEQPEQP
	gliadin	100% match			(DQ2.2 and
		to bacterial			DQ2.5)
		protein and
		lizard protein
		lower to
		grapes and
		wheat Glia-
		gamma2 in
		Deamidated
		form
472	gamma-	CAUTION	Immunogenic	Native	DQ2.5/	90, 17	9	QQPQQPFPQ
	gliadin	100% 4			DQ8	8
		matches to
		Candida 2 to
		bacteria and
		many to
		secalins and
		wheat
		gamma2-
		gliadin
473	gamma-2	CAUTION	Immunogenic	Deamidated	DQ2.5/	90, 17,	9	EQPQQPFPQ
	gliadin	100% match			DQ8	8
		to fungal
		protein and
		lower to
		Candida and
		to Gliadin
		epitope:
		gamma-l
474	gamma-2	Gliadin	Immunogenic	Deamidated	DQ2.5/	90, 17,	9	QQPEQPFPQ
	gliadin	epitope:			DQ8	8
		gamma-VII
475	gamma-2	CAUTION	Immunogenic	Deamidated	DQ2.5/	25, 90,	9	EQPEQPFPQ
	gliadin	100% match			DQ8	17
		to a bacterial
		protein /
		lower to
		triticum
		gammaVII-
		gliadin in
		Deamidated
		form
476	gamma-	gamma-Glia	Immunogenic	Native	DQ2	57	14	PQQQTLQPQQPAQ
	gliadin	(p105-p118)						L
477	gamma-	gamma-Glia	Immunogenic	Deamidated	DQ2	57	14	PQQQTLQPEQPAQ
	gliadin	(p105-p118;						L
		E113)
478	gamma1-	Wheat	Immunogenic	Native	DQ2	62	20	ATANMQVDPSGQ
	gliadin	peptide W37						VQWPQQQP
479	gamma1-	Wheat	Immunogenic	Native	DQ2	62	12	QVDPSGQVQWPQ
	gliadin	peptide W37
480	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	17, 23	20	WPQQQPFPQPQQ
	gliadin	1370 (p1-p30);						PFCQQPQR
		gamma-
		gliadin M2
		M36999 (p11-
		p30)
		homologous
		to DQ2-alpha-I
481	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2	17	20	QQPFCQQPQRTIP
	gliadin	1371 (p21-						QPHQTFH
		p40)
482	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2	17	20	TIPQPHQTFHHQPQ
	gliadin	1372 (p31-						QTFPQP
		p50)
483	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	17	20	HQPQQTFPQPQQT
	gliadin	1372 (p41-						YPHQPQQ
		p60)
484	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2	17	20	QQTYPHQPQQQFP
	gliadin	1372 (p51-						QTQQPQQ
		p70)
485	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	25, 17,	20	QFPQTQQPQQPFP
	gliadin	1375 (p61-				23		QPQQTFP
		p80); gamma-
		gliadin M7
		M36999 (p61-
		p80)
		homologous
		to DQ2-
		gamma-III
486	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETQQPQQPFP
	gliadin	1375 (p61-						QPQQTFP
		p80; E64);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
487	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPQTEQPQQPFP
	gliadin	1375 (p61-						QPQQTFP
		p80; E66);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
488	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPQTQQPEQPFP
	gliadin	1375 (p61-						QPQQTFP
		p80; E69);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
489	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPQTQQPQQPFP
	gliadin	1375 (p61-						QPEQTFP
		p80; E76);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
490	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETEQPQQPFP
	gliadin	1375 (p61-						QPQQTFP
		p80; E64 and
		E66); gamma-
		gliadin M7
		M36999 (p61-
		p80)
		homologous
		to DQ2-
		gamma-III
491	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETQQPEQPFP
	gliadin	1375 (p61-						QPQQTFP
		p80; E64 and
		E69); gamma-
		gliadin M7
		M36999 (p61-
		p80)
		homologous
		to DQ2-
		gamma-III
492	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETQQPQQPFP
	gliadin	1375 (p61-						QPEQTFP
		p80; E64 and
		E76); gamma-
		gliadin M7
		M36999 (p61-
		p80)
		homologous
		to DQ2-
		gamma-III
493	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPQTEQPEQPFP
	gliadin	1375 (p61-						QPQQTFP
		p80; E66 and
		E69); gamma-
		gliadin M7
		M36999 (p61-
		p80)
		homologous
		to DQ2-
		gamma-III
494	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPQTEQPQQPFP
	gliadin	1375 (p61-						QPEQTFP
		p80; E66 and
		E76); gamma-
		gliadin M7
		M36999 (p61-
		p80)
		homologous
		to DQ2-
		gamma-III
495	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPQTQQPEQPFP
	gliadin	1375 (p61-						QPEQTFP
		p80; E69 and
		E76); gamma-
		gliadin M7
		M36999 (p61-
		p80)
		homologous
		to DQ2-
		gamma-III
496	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETEQPEQPFPQ
	gliadin	1375 (p61-						PQQTFP
		p80; E64, E66
		and E69);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
497	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETEQPQQPFP
	gliadin	1375 (p61-						QPEQTFP
		p80; E64, E66
		and E76);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
498	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETQQPEQPFP
	gliadin	1375 (p61-						QPEQTFP
		p80; E64, E69
		and E76);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
499	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPQTEQPEQPFP
	gliadin	1375 (p61-						QPEQTFP
		p80; E66, E69
		and E76);
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
500	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	QFPETEQPEQPFPQ
	gliadin	1375 (p61-						PEQTFP
		p80; E64, E66,
		E69 and E76) ;
		gamma-gliadin
		M7 M36999
		(p61-p80)
		homologous
		to DQ2-
		gamma-III
501	gamma1-	Wheat	Immunogenic	Native	DQ2	62	20	PQQPFPQPQQTFP
	gliadin	peptide W28,						QQPQLPF
		W33
502	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	17, 23	20	PFPQPQQTFPQQP
	gliadin	1376 (p71-						QLPFPQQ
		p90); gamma-
		gliadin M8
		M36999 (71-80)
		homologous
		to DQ2-alpha-I
		and DQ2-
		gamma-IV
503	gamma1-	Wheat	Immunogenic	Native	DQ2	62	11	PFPQPQQTFPQ
	gliadin	peptide W33
504	gamma1-	Wheat	Immunogenic	Native	DQ2	62	12	PQQTFPQQPQLP
	gliadin	peptide W28
505	gamma1-	Wheat	Immunogenic	Native	DQ2	62	20	SQQPQQQFSQPQQ
	gliadin	peptide W10						QFPQPQQ
506	gamma1-	DQ2-y-IV y-	Immunogenic	Native	DQ2	57, 23	15	QQFSQPQQQFPQP
	gliadin	Glia (p117-						QQ
		p132)
507	gamma1-	DQ2-y-IV y-	Immunogenic	Deamidated	DQ2	57, 23	15	QQFSQPEQQFPQP
	gliadin	Glia (p117-						QQ
		p132; E123)
508	gamma1-	DQ2-y-IV y-	Immunogenic	Deamidated	DQ2	57, 23	15	QQFSQPQQEFPQP
	gliadin	Glia (p117-						QQ
		p132; E125)
509	gamma1-	DQ2-y-IV y-	Immunogenic	Deamidated	DQ2	57, 23	15	QQFSQPEQEFPQP
	gliadin	Glia (p117-						QQ
		p132; E123
		and E125)
510	gamma1-	Wheat	Immunogenic	Native	DQ2	62	12	QQFSQPQQQFPQ
	gliadin	peptide W10
511	gamma1-	gamma-IV	Immunogenic	Native	DQ2	23	13	QFSQPQQQFPQPQ
	gliadin	(p101-p113)
512	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2	23	12	FSQPQQQFPQPQ
	gliadin	(gamma5
		p102-p113)
513	gamma1-	gamma5	Immunogenic	Deamidated	DQ2	23	12	FSQPEQQFPQPQ
	gliadin	(p102-p113;
		E106)
514	gamma1-	gamma5	Immunogenic	Deamidated	DQ2	23	12	FSQPQQEFPQPQ
	gliadin	(p102-p113;
		E108)
515	gamma1-	gamma5	Immunogenic	Deamidated	DQ2	25,23	12	FSQPEQEFPQPQ
	gliadin	(p102-p113;
		E106 and
		E108)
516	gamma1-	gamma5	Immunogenic	Native	DQ2	25	10	FSQPQQQFPQ
	gliadin	(p102-p111)
517	gamma1-	gamma5	Immunogenic	Deamidated	DQ2	25	10	FSQPEQQFPQ
	gliadin	(p102-p111;
		E106)
518	gamma1-	gamma5	Immunogenic	Deamidated	DQ2	25	10	FSQPQQEFPQ
	gliadin	(p102-p111;
		E108)
519	gamma1-	gamma5	Immunogenic	Deamidated	DQ2	25	10	FSQPEQEFPQ
	gliadin	(p102-p111;
		E106 and
		E108
520	gamma1-	gamma-IV	Immunogenic	Native	DQ2	23	12	SQPQQQFPQPQQ
	gliadin	(p103-p114)
521	gamma-5	CAUTION	Immunogenic	Native	DQ2.5	90, 25,	9	SQPQQQFPQ
	gliadin	100% match				23
		to fungal
		protein and
		many wheat
		proteins glia-
		gamma 4a
522	gamma-5	glia-gamma 4a	Immunogenic	Deamidated	DQ2.5	90, 25,	9	SQPEQEFPQ
	gliadin					23
523	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2	24	11	QQPQQSFPQQQ
	gliadin	of
		GDB2_WHEAT
		(SwissProt
		P08453
		GI:121101)
		(p140-p150)
524	gamma1-	gamma-gliadin	Immunogenic	Native	DQ2	24	10	QPQQSFPQQQ
	gliadin	of
		GDB2 WHEAT
		(SwissProt
		P08453
		GI170738)
		(p141-p150)
525	gamma1-	gamma-gliadin	Immunogenic	Deamidated	DQ2	24	10	QPQQSFPEQQ
	gliadin	of
		GDB2_WHEAT
		(SwissProt
		P08453
		GI170738)
		(p141-p150;
		E148)
526	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	WPQQQPFPQPQQ
	gliadin	peptide W07						PFCQQPQQ
527	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	20	WPQQQPFPQPEQP
	gliadin	peptide W07						FCQQPQQ
529	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPFCQ
	gliadin	peptide W07
530	gamma-	Predicted	Immunogenic	Native	DQ8	22	14	QFPQTQQPQQPFP
	gliadin	gamma-gliadin			(DQ2/8)			Q
531	gamma-	gamma-gliadin	Immunogenic	Synthesised	DQ8	17	14	PQTEQPQQPFPQP
	gliadin	M36999 (p63-		as				Q
		p76; E66)		Deamidated
532	gamma-	gamma-gliadin	Immunogenic	Synthesised	DQ2	17	14	PQTQQPEQPFPQP
	gliadin	M36999 (p63-		as				Q
		p76; E69)		Deamidated
533	gamma-	gamma-gliadin	Immunogenic	Synthesised	DQ2, DQ8	17	14	PQTEQPEQPFPQP
	gliadin	M36999 (p63-		as				Q
		p76; E66 and		Deamidated
		E69)
534	gamma-	gamma-gliadin	Immunogenic	Native	DQ2	25	11	TOOPQQPFPQP
	gliadin	1375 (p61-
		p80); gamma-
		gliadin M7
		M36999 (61-
		80)
		homologous
		to DQ2-
		gamma-III
535	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	11	TEQPEQPFPQP
	gliadin	1375 (p61-
		p80; E62 and
		E65),;
		gamma-gliadin
		M7 M36999
		(61-80)
		homologous
		to DQ2-
		gamma-III
536	gamma-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	17	20	QQPQLPFPQQPQQ
	gliadin	1377 (p81-						PFPQPQQ
		p100)
537	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	22	14	QLPFPQQPQQPFP
	gliadin	(p84-p97)			(DQ2/8)			Q
538	gamma-	Glia-gamma2	Immunogenic	Native	DQ2	20	10	PFPQQPQQPF
	gliadin	(p89-p102)
539	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	20	10	PFPEQPQQPF
	gliadin	(p89-p102;
		E92)
540	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	20	10	PFPQQPEQPF
	gliadin	(p89-p102;
		E94)
541	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	20	10	PFPEQPEQPF
	gliadin	(p89-p102;
		E92 and E94)
542	gamma-	CAUTION	Immunogenic	Native	DQ2	27	9	FPQQPQQPF
	gliadin	100% match
		to a fungal
		protein and
		many Secalins
		gamma-
		Gliadin (p90-
		p102)
543	gamma-	gamma-	Immunogenic	Deamidated	DQ2	27	9	FPEQPQQPF
	gliadin	Gliadin (p90-
		p102; E92)
544	gamma-	gamma-	Immunogenic	Deamidated	DQ2	27	9	FPQQPQEPF
	gliadin	Gliadin (p90-
		p102; E96)
545	gamma-	gamma-	Immunogenic	Deamidated	DQ2	27	9	FPEQPQEPF
	gliadin	Gliadin (p90-
		p102; E92 and
		E96)
546	gamma-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	17, 23	20	PQQPFPQPQQPQQ
	gliadin	1378 (p91-						PFPQSQQ
		p110),;
		gamma-gliadin
		M10 M36999
		(91-110)
		homologous
		to DQ2-alpha-I
547	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	QQPAQYEVIRSLVL
	gliadin	peptide W36						RTLPNM
548	gamma-	Wheat	Immunogenic	Native	DQ2	62	16	QYEVIRSLVLRT
	gliadin	peptide W36						LPNM
549	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	15	EYEVIRSLVLRT
	gliadin	peptide W36						LPN
550	gamma-	Wheat	Immunogenic	Native	DQ2	62	15	QYQVIRSLVLRT
	gliadin	peptide W36						LPN
551	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	54	20	QQQFIQPQQPFPQ
	gliadin	AAK84778						QPQQTYP
		(p74-p93)
552	gamma-	Wheat	Immunogenic	Native	DQ2	62	12	QQFIQPQQPFPQ
	gliadin	peptide W14
553	gamma-	Predicted	Immunogenic	Native	DQ8	22	14	PFPQTQQPQQPFP
	gliadin	gamma-gliadin			(DQ2/8)			Q
554	gamma-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	17	20	PQQPFPQSQQPQQ
	gliadin	1379 (p101-						PFPQPQQ
		p120)
555	gamma-	Predicted	Immunogenic	Native	DQ8	22	14	PFPQSQQPQQPFP
	gliadin	gamma-gliadin			(DQ2/8)			Q
556	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	SQQPQQPFPQPQQ
	gliadin	peptide W16						QFPQPQQ
557	gamma-	gamma-gliadin	Immunogenic	Native	DQ2	25, 17,	20	PQQPFPQPQQQFP
	gliadin	1380 (p111-				23		QPQQPQQ
		p130);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
558	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPQQQFP
	gliadin	1380 (p111-						QPQQPQQ
		p130; E112) ;
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
559	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PQQPFPQPEQQFP
	gliadin	1380 (p111-						QPQQPQQ
		p130; E119);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
560	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PQQPFPQPQQEFP
	gliadin	1380 (p111-						QPQQPQQ
		p130; E121);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
561	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PQQPFPQPQQQFP
	gliadin	1380 (p111-						QPEQPQQ
		p130; E126);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
562	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPEQQFP
	gliadin	1380 (p111-						QPQQPQQ
		p130; E112
		and E119);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
563	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPQQEFP
	gliadin	1380 (p111-						QPQQPQQ
		p130; E112
		and E121);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
564	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPQQQFP
	gliadin	1380 (p111-						QPEQPQQ
		p130; E112
		and E126);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
565	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PQQPFPQPEQEFP
	gliadin	1380 (p111-						QPQQPQQ
		p130; E119
		and E121);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
566	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PQQPFPQPEQQFP
	gliadin	1380 (p111-						QPEQPQQ
		p130; E119
		and E126);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
567	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PQQPFPQPQQEFP
	gliadin	1380 (p111-						QPEQPQQ
		p130; E121
		and E126);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
568	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPEQEFPQ
	gliadin	1380 (p111-						PQQPQQ
		p130; E112,
		E119 and
		E121); gamma
		gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
569	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPEQQFP
	gliadin	1380 (p111-						QPEQPQQ
		p130; E112,
		E119 and
		E126); gamma-
		gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
570	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPQQEFP
	gliadin	1380 (p111-						QPEQPQQ
		p130; E112,
		E121 and
		E126); gamma-
		gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
571	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PQQPFPQPEQEFP
	gliadin	1380 (p111-						QPEQPQQ
		p130; E119,
		E121 and
		E126); gamma-
		gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
572	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	20	PEQPFPQPEQEFPQ
	gliadin	1380 (p111-						PEQPQQ
		p130; E112,
		E119, E121
		and E126);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
573	gamma-	gamma-gliadin	Immunogenic	Native	DQ2	62, 25	12	FPQPQQQFPQPQ
	gliadin	1380 (p111-
		p130);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV;
		W16
574	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	12	FPQPEQQFPQPQ
	gliadin	1380 (p111-
		p130; E115);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
575	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	12	FPQPQQEFPQPQ
	gliadin	1380 (p111-
		p130; E117);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
576	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	12	FPQPEQEFPQPQ
	gliadin	1380 (p111-
		p130; E115
		and E117);
		gamma-gliadin
		M12 M36999
		(111-130)
		homologous
		to DQ2-
		gamma-IV
577	gamma-	CAUTION	Immunogenic	Native	DQ2.5	90,25	9	PQPQQQFPQ
	gliadin	100% matches
		to 3 fungal
		and metazoan
		proteins and
		wheat glia-
		gamma 4b
578	gamma-	glia-gamma 4b	Immunogenic	Deamidated	DQ2.5	90, 25	9	PQPEQQFPQ
	gliadin
579	gamma-	glia gamma 4b	Immunogenic	Deamidated	DQ2.5	90, 25	9	PQPQQEFPQ
	gliadin
580	gamma-	glia-gamma 4b	Immunogenic	Deamidated	DQ2.5	90, 25	9	PQPEQEFPQ
	gliadin
581	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	16	GQQPFPQPEQEFP
	gliadin	peptide W16						QPG
582	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	13	QPFPQPEQEFPQP
	gliadin	peptide W16
583	gamma-1	glia-alpha 1,	Immunogenic	Native	DQ2.5/	8, 74,	9	PQQSFPQQQ
	gliadin	glia-gamma 1			DQ8	76, 24
584	gamma-	glia-alpha1,	Immunogenic	Deamidated	DQ2.5/	8, 74,	9	PQQSFPQQE
	gliadin	glia-gamma1			DQ8	76, 24
585	gamma-1	glia-alpha 1,	Immunogenic	Deamidated	DQ2.5/	8, 76,	9	PQQSFPEQE
	gliadin	glia-gamma 1			DQ8	24
586	gamma-1	glia-alpha1,	Immunogenic	Deamidated	DQ2.5,	8, 17,	9	PQQSFPEQQ
	gliadin	glia-gamma1			DQ8	76, 24
587	gamma-	Glia-	Immunogenic	Native	DQ2	19	15	VQGQGIIQPQQPA
	gliadin	gamma30-
		gliadin (p222-
		p236)
588	gamma-	Glia-	Immunogenic	Deamidated	DQ2	19	15	VQGEGIIQPQQPAQ
	gliadin	gamma30-						L
		gliadin (p222-
		236; E225)
589	gamma-	Glia-	Immunogenic	Deamidated	DQ2	19	15	VQGQGIIQPEQPAQ
	gliadin	gamma30-						L
		gliadin (p222-
		236; E231)
590	gamma-	Glia-	Immunogenic	Deamidated	DQ2	19	15	VQGEGIIQPEQPAQ
	gliadin	gamma30-						L
		gliadin (p222-
		236; E225 and
		E231)
591	gamma-	DQ2-y-II y-	Immunogenic	Native	DQ2	57	15	GQGIIQPQQPAQLI
	gliadin	Glia (p222-						R
		p236)
592	gamma-	DQ2-y-II y-	Immunogenic	Deamidated	DQ2	57	15	GQGIIQPEQPAQL
	gliadin	Glia (p222-						IR
		p236; E229)
593	gamma-	gamma5-	Immunogenic	Native	DQ2	25	11	GIIQPQQPAQL
	gliadin	gliadin (p227-
		p237) ;
		gamma-II
		epitope
594	gamma-	gamma5-	Immunogenic	Deamidated	DQ2	25	11	GIIQPEQPAQL
	gliadin	gliadin (p227-
		237; E232)
595	gamma-	gamma5-	Immunogenic	Native	DQ2	47, 25	10	IIQPQQPAQL
	gliadin	gliadin (p228-
		237)
596	gamma-	gamma5-	Immunogenic	Deamidated	DQ2	47, 25	10	IIQPEQPAQL
	gliadin	gliadin (p228-
		237; E232)
597	gamma-	gamma-2	Immunogenic	Native	DQ2	19, 23,	9	IIQPQQPAQ
	gliadin	peptide 1306;				2
		Glia-
		gamma30-
		gliadin (p227-
		p235) minimal
		epitope
598	gamma-	gamma-2	Immunogenic	Deamidated	DQ2	19	9	IIQPEQPAQ
	gliadin	peptide 1306;
		Glia-
		gamma30-
		gliadin (p228-
		p235; E232)
		minimal
		epitope
599	gamma-5	glia-gamma 2	Immunogenic	Native	DQ2.5	90, 8,	9	IQPQQPAQL
	gliadin					25
600	gamma-5	glia-gamma 2	Immunogenic	Deamidated	DQ2.5	90, 8,	9	IQPEQPAQL
	gliadin					25
601	gamma-	gamma-gliadin	Immunogenic	Native	DQ2, DQ8	17, 23	20	QFPQPQQPQQSFP
	gliadin	1381 (p121-						QQQQPAI
		p140);
		gamma-gliadin
		M13 M36999
		(121-140)
		identical to
		DQ2-gamma-I
602	gamma-	DQ2-gamma-I	Immunogenic	Native	DQ2	57	15	PQQPQQSFPQQQ
	gliadin	gamma-Glia						QPA
		(p139-p153)
603	gamma-	DQ2-gamma-I	Immunogenic	Deamidated	DQ2	57	15	PQQPQQSFPEQQQ
	gliadin	gamma-Glia						PA
		(p139-p153;
		E147)
604	gamma-	DQ2-gamma-I	Immunogenic	Deamidated	DQ2	57	15	PQQPQQSFPQQEQ
	gliadin	gamma-Glia						PA
		(p139-p153;
		E150)
605	gamma-	DQ2-gamma-I	Immunogenic	Deamidated	DQ2	57	15	PQQPQQSFPEQEQ
	gliadin	gamma-Glia						PA
		(p139-p153;
		E147 and
		E150)
606	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	17	20	SFPQQQQPAIQSFL
	gliadin	1382 (p131-						QQQMNP
		p150)
607	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	17	20	QSFLQQQMNPCKN
	gliadin	1383 (p141-						FLLQQCN
		p160)
608	gamma-	gamma-gliadin	Immunogenic	Native	DQ2	17	20	IHSVAHSIIMQQEQ
	gliadin	1388 (p201-						QQGVPI
		p220)
609	gamma-	gamma-gliadin	Immunogenic	Native	DQ2	17, 23	20	LRPLFQLAQGLG
	gliadin	M23 M36999						IIQPQQPA
		(221-240)
		homologous
		to DQ2-
		gamma-II
610	gamma-	gamma-gliadin	Immunogenic	Native	DQ2	17, 23	20	LGIIQPQQPAQL
	gliadin	1391 (p231-						EGIRSLVL
		p250) ;
		gamma-gliadin
		M24 M36999
		(231-250)
		identical to
		DQ2-gamma-II
611	gamma-	Gluten	Immunogenic	Native	DQ2	61	18	PHQPQQQVPQPQ
	gliadin	peptide #19			(DQ2.5)			QPQQPF
612	gamma-	Predicted	Immunogenic	Native	DQ8	22, 8	14	QQPFPQQPQQPFP
	gliadin	gamma-gliadin			(DQ2/8)			Q
613	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	8	14	QQPFPEQPEQPFP
	gliadin	in Deamidated						Q
		form
614	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	8	14	QQPFPQQPEQPFP
	gliadin	in Deamidated						Q
		form
615	gamma-	Glia-gamma2	Immunogenic	Deamidated	DQ2	8	14	QQPFPEQPQQPFP
	gliadin	in Deamidated						Q
		form
616	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ8	54	20	QTQQPQQPFPQQP
	gliadin	P08453 (p94-						QQPFPQT
		p113)
617	gamma-	Predicted	Immunogenic	Native	DQ8	22	14	PFPQLQQPQQPFP
	gliadin	gamma-gliadin			(DQ2/8)			Q
618	gamma-	gamma-I	Immunogenic	Native	DQ2	2	21	YQQLPQPQQPQQS
	gliadin	gliadin 1206						FPQQQRPF
619	gamma-	gamma-type	Immunogenic	Deamidated	DQ2	24	20	QQLPQPQQPQQSF
	gliadin	gliadin of						PQQQRPF
		GDB2_WHEAT
		(SwissProt
		P08453)
		(p134-p153)
620	gamma-	Glia-gamma1	Immunogenic	Native	DQ2	9	17	QPQQPQQSFPQQ
	gliadin	epitope						QRPFI
621	gamma-	Glia-	Immunogenic	Native	DQ2	19	16	QPQQPQQSFPQQ
	gliadin	gamma1 (p138-						QRPF
		p153)
622	gamma-	Glia-gamma1	Immunogenic	Native	DQ2	8	15	PQQPQQSFPQQQR
	gliadin	(p139-p153)						PF
623	gamma-	Glia-gamma1	Immunogenic	Deamidated	DQ2	8	15	PQQPQQSFPEQQR
	gliadin	(p139-p153;						PF
		E148)
624	gamma-	Glia-gamma1	Immunogenic	Deamidated	DQ2	8	15	PEQPQQSFPEQQR
	gliadin	(p139-p153;						PF
		E140 and
		E148)
625	gamma-	Glia-gamma1	Immunogenic	Deamidated	DQ2	8	15	PQQPQQSFPEQER
	gliadin	(p139-p153;						PF
		E148 and
		E150)
626	gamma-	Glia-gamma1	Immunogenic	Deamidated	DQ2	8	15	PEQPQQSFPEQERP
	gliadin	(p139-p153;						F
		E140, E148
		and E150)
627	gamma-	gamma-I,	Immunogenic	Native	DQ2	43, 25	14	PQQPQQSFPQQQR
	gliadin	gamma-						P
		Gliadin (p139-
		p152)
628	gamma-	gamma-	Immunogenic	Deamidated	DQ2	43, 25	14	PEQPQQSFPEQERP
	gliadin	Gliadin (p139-
		p152; E140,
		E148 and
		E150) E
		residues in the
		gliadin
		peptides are
		introduced to
		mimic the
		deamidation
		mediated by
		tissue
		trans-
		glutaminase.
629	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	55	14	PQQPQQSFPEQQR
	gliadin	(p139-p152;						P
		E148)
630	gamma-	gamma-gliadin	Immunogenic	Deamidated	DQ2	25	14	PEQPQQSFPEQQR
	gliadin	(p139-p152;			(DQ2.2 and			P
		E140 and			DQ2.5)
		E148)
631	gamma-	P-3 gamma-	Immunogenic	Synthesised	DQ2	56	14	KEQPQQSFPEQERP
	gliadin	gliadin (p139-		as
		p152; K139,		Deamidated
		E140, E148
		and E150)
632	gamma-	P-2 gamma-	Immunogenic	Synthesised	DQ2	56	14	PKQPQQSFPEQERP
	gliadin	gliadin (p139-		as
		p152; K140,		Deamidated
		E148 and
		E150)
633	gamma-	P-1 gamma-	Immunogenic	Synthesised	DQ2	56	14	PEKPQQSFPEQERP
	gliadin	gliadin (p139-		as
		p152; K141,		Deamidated
		E140, E148
		and E150)
634	gamma-	P1 γ-	Immunogenic	Synthesised	DQ2	56	14	PEQKQQSFPEQERP
	gliadin	gliadin(p139-		as
		p152; K142,		Deamidated
		E140, E148
		and E150)
635	gamma-	P2 γ-gliadin	Immunogenic	Synthesised	DQ2	56	14	PEQPKQSFPEQERP
	gliadin	(p139-p152;		as
		K143, E140,		Deamidated
		E148 and
		E150)
636	gamma-	P4 γ-gliadin	Immunogenic	Synthesised	DQ2	56	14	PEQPQQKFPEQERP
	gliadin	(p139-p152;		as
		K144, E140,		Deamidated
		E148 and
		E150)
637	gamma-	P9 gamma-	Immunogenic	Synthesised	DQ2	56	14	PEQPQQSFPEQKRP
	gliadin	gliadin (p139-		as
		p152; E140,		Deamidated
		E148 and
		K150)
638	gamma-	P10 gamma-	Immunogenic	Synthesised	DQ2	56	14	PEQPQQSFPEQEKP
	gliadin	gliadin (p139-		as
		p152; K151,		Deamidated
		E140, E148
		and E150)
639	gamma-	P11 γ-gliadin	Immunogenic	Synthesised	DQ2	56	14	PEQPQQSFPEQERK
	gliadin	(p139-p152;		as
		K152, E140,		Deamidated
		E148 and
		E150)
640	gamma-	gamma-I	Immunogenic	Native	DQ2	47	12	QPQQSFPQQQRP
	gliadin	epitope in
		native form
641	gamma-	Deamidated	Immunogenic	Deamidated	DQ2	47	12	QPQQSFPEQQRP
	gliadin	form of
		gamma-I
		epitope
642	gamma-	Glia-alpha20	Immunogenic	Native	DQ2	9	16	QQSFPQQQRPFIQP
	gliadin							SI
643	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	54	20	PQPQQPQLPFPQQ
	gliadin	AAK84772						PQQPFPQ
		(p130-p149)
644	gamma-	Predicted	Immunogenic	Native	DQ8	22	14	PFPQPQQPQQPFP
	gliadin	gamma-gliadin			(DQ2/8)			Q
645	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	54	20	QQPLPQPQQPQQP
	gliadin	AAK84776						FPQSQQP
		(p102-p121)
646	gamma-	gamma-gliadin	Immunogenic	Native	DQ8	54	20	QPQQPQQPFPQQ
	gliadin	AAK84772						QQPLIQPY
		(p121-p140)
647	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	PQQPFPQQPQQQF
	gliadin	peptide W35						PQPQQPQ
648	gamma-	Wheat	Immunogenic	Native	DQ2	62	12	PFPQQPQQQFPQ
	gliadin	peptide W35
649	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	QPFPQLQQPQQPL
	gliadin	peptide W31						PQPQQPQ
650	gamma-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQLQQPQQP
	gliadin	peptide W31
651	LMW	Wheat	Immunogenic	Native	DQ2	62	20	SHIPGLERPWQQQ
	glutenin	peptide W15						PLPPQQT
		LMW
652	LMW	Wheat	Immunogenic	Native	DQ2	62	15	QGLERPWQQQPLP
	glutenin	peptide W15						PQ
		LMW
653	LMW	Wheat	Immunogenic	Deamidated	DQ2	62	15	EGLERPWQEQPLPP
	glutenin	peptide W15						Q
		LMW
654	LMW	Wheat	Immunogenic	Native	DQ2	62	12	LERPWQQQPLPP
	glutenin	peptide W15
		LMW
655	LMW	Wheat	Immunogenic	Deamidated	DQ2	62	16	GQQAFPQPEQTFP
	glutenin	peptide W11						HQG
656	LMW	Wheat	Immunogenic	Native	DQ2	62	15	QQAFPQPQQTFPH
	glutenin	peptide W11						QP
657	LMW	Wheat	Immunogenic	Deamidated	DQ2	62	15	EQAFPQPEQTFPH
	glutenin	peptide W11						QP
658	LMW	Wheat	Immunogenic	Native	DQ2	62	20	QAFPQPQQTFPHQ
	glutenin	peptide W11						PQQQFPQ
659	LMW	Wheat	Immunogenic	Native	DQ2	62	12	QAFPQPQQTFPH
	glutenin	peptide W11
660	LMW	Wheat	Immunogenic	Deamidated	DQ2	62	12	QAFPQPEQTFPH
	glutenin	peptide W11
661	gamma-	Glutenin-Glt-	Immunogenic	Native	DQ2	19	15	QQPPFSQQQQQPL
	gliadin	17 (p46-p60)						PQ
	or LMW
	glutenin
662	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEQQQQPL
	gliadin	17 (p46-p60;						PQ
	or LMW	E52)
	glutenin
663	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSQEQQQPL
	gliadin	17 (p46-p60;						PQ
	or LMW	E53)
	glutenin
664	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSQQQEQPL
	gliadin	17 (p46-p60;						PQ
	or LMW	E55)
	glutenin
665	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSQQQQEPL
	gliadin	17 (p46-p60;						PQ
	or LMW	E56)
	glutenin
666	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEEQQQPLP
	gliadin	17 (p46-p60;
	or LMW	E52 and 53)
	glutenin
667	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEQQEQPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E52 and 55)
	glutenin
668	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEQQQEPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E52 and 56)
	glutenin
669	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSQEQEQPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E53 and 55)
	glutenin
670	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSQEQQEPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E53 and 56)
	glutenin
671	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSQQQEEPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E55 and 56)
	glutenin
672	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEEQEQPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E52, 53 and
	glutenin	55)
673	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEEQQEPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E52, 53 and
	glutenin	56)
674	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSQEQEEPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E53, 55 and
	glutenin	56)
675	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEQQEEPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E52, 55 and
	glutenin	56)
676	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	19	15	QQPPFSEEQEEPLP
	gliadin	17 (p46-p60;						Q
	or LMW	E52, 53, 55
	glutenin	and 56)
693	gamma-	CAUTION	Immunogenic	Native	DQ2	27	9	FSQQQQQPL
	gliadin	100% matches
	or LMW	to 5 microbial
	glutenin	proteins and
		to wheat
		proteins
		Glutenin-Glt-
		17 (p50-p58)
694	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	27	9	FSEQQQQPL
	gliadin	17 (p50-p58;
	or LMW	E52)
	glutenin
695	gamma-	CAUTION	Immunogenic	Deamidated	DQ2	27	9	FSQEQQQPL
	gliadin	100% matches
	or LMW	to two Pinus
	glutenin	proteins/not
		wheat
		Glutenin-Glt-
		17 (p50-p58;
		E53)
696	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	27	9	FSQQQEQPL
	gliadin	17 (p50-p58;
	or LMW	E55)
	glutenin
697	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	27	9	FSEEQQQPL
	gliadin	17 (p50-p58;
	or LMW	E52 and E53)
	glutenin
698	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	27	9	FSEQQEQPL
	gliadin	17 (p50-p58;
	or LMW	E52 and E55)
	glutenin
699	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	27	9	FSQEQEQPL
	gliadin	17 (p50-p58;
	or LMW	E53 and E55)
	glutenin
700	gamma-	Glutenin-Glt-	Immunogenic	Deamidated	DQ2	27	9	FSEEQQEPL
	gliadin	17 (p50-p58;
	or LMW	E52, E53 and
	glutenin	E55)
701	gamma-	Glutenin-17	Immunogenic	Native	DQ2	8, 19	15	QQPPFSQQQQPVL
	gliadin	epitope						PQ
	or LMW	homolog
	glutenin
702	gamma-	Glutenin-17	Immunogenic	Deamidated	DQ2	8	15	QQPPFSEQQQPVL
	gliadin	epitope						PQ
	or LMW	homolog in
	glutenin	Deamidated
		form
703	gamma-	Glutenin-17	Immunogenic	Deamidated	DQ2	8	15	QQPPFSQQEQPVL
	gliadin	epitope						PQ
	or LMW	homolog in
	glutenin	Deamidated
		form
704	gamma-	Glutenin-17	Immunogenic	Deamidated	DQ2	8	15	QQPPFSEQEQPVLP
	gliadin	epitope						Q
	or LMW	homolog in
	glutenin	Deamidated
		form
705	gamma-	LMW T cell	Immunogenic	Deamidated	DQ2	62	15	EQPPFSEQEQPVLP
	gliadin	epitope						Q
	or LMW
	glutenin
706	Glut-L1	CAUTION	Immunogenic	Native	DQ2.2	44, 90	9	PFSQQQQPV
		100% match
		to a fungal
		protien
		Hebeloma sp.
		& many wheat
		Glt-17 (Var1)
707	glut-L1	Glt-17 (Var1)	Immunogenic	Deamidated	DQ2.2	44, 90	9	PFSEQQQPV
708	glut-L1	Glt-17 (Var1)	Immunogenic	Deamidated	DQ2.2	44, 90	9	PFSQQEQPV
709	glut-L1	Glt-17 (Var1)	Immunogenic	Deamidated	DQ2.2	44, 90	9	PFSEQEQPV
710	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	CKVFLQQQCSPVA
	gliadin	peptide W12						MPQRLAR
	or LMW
	glutenin
711	gamma-	Wheat	Immunogenic	Native	DQ2	62	16	LQQQCSPVAMPQR
	gliadin	peptide W12						LAR
	or LMW
	glutenin
712	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	PQQQQPFPQPQQP
	gliadin	peptide W05						FSQQPQQ
	or LMW
	glutenin
713	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	20	PQQQQPFPQPEQP
	gliadin	peptide W05						FSQQPQQ
	or LMW
	glutenin
714	gamma-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQPQQPFSQ
	gliadin	peptide W05
	or LMW
	glutenin
715	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPFSQ
	gliadin	peptide W05
	or LMW
	glutenin
716	gamma-	Wheat	Immunogenic	Native	DQ2	62	20	QQPFPQPQQPQLP
	gliadin	peptide W17						FPQQPQQ
	or LMW
	glutenin
717	gamma-	Wheat	Immunogenic	Native	DQ2	62	15	QPFPQPQQPQLPFP
	gliadin	peptide W17						Q
	or LMW
	glutenin
718	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	15	EPFPQPEQPELPFP
	gliadin	peptide W17						Q
	or LMW
	glutenin
719	gamma-	Wheat	Immunogenic	Native	DQ2	62	12	QPFPQPQQPQLP
	gliadin	peptide W17
	or LMW
	glutenin
720	LMW	GLT/GLIA	Immunogenic	Native	DQ2	19	15	QQPPFSQQQQPPF
	glutenin	homologue						SQ
		peptide 12
721	LMW	LMW glutenin-	Immunogenic	Native	DQ2	19	20	QQQQPPFSQQQQS
	glutenin	glt-156 (p40-						PFSQQQQ
		p59)
722	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	19	20	QQQQPPFSEQQQS
	glutenin	glt-156 (p40-						PFSQQQQ
		p59; E48)
723	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	19	20	QQQQPPFSQQQES
	glutenin	glt-156 (p40-						PFSQQQQ
		p59; E51)
724	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	19	20	QQQQPPFSEQQES
	glutenin	glt-156 (p40-						PFSQQQQ
		p59; E48 and
		E51)
725	LMW	Homolog of	Immunogenic	Deamidated	DQ2	20	15	QQQQPPFSEEQESP
	glutenin	Deamidated						Y
		Glt-156
		minimal
		epitope (p40-
		p59)
726	LMW	Homolog of	Immunogenic	Deamidated	DQ2	20	15	QQQQPPFSEEQESP
	glutenin	Deamidated						L
		Glt-156
		minimal
		epitope (p40-
		p59)
727	LMW	Deamidated	Immunogenic	Deamidated	DQ2	20	15	QQQQPPFSEEQESP
	glutenin	Glt-156						F
		minimal
		epitope (p40-
		p59)
728	LMW	Glt-156	Immunogenic	Deamidated	DQ2	20	15	QQQPPFSEEQESPF
	glutenin	minimal						S
		epitope (p41-
		p55)
729	LMW	Glt-156	Immunogenic	Native	DQ2	20	15	QQPPFSQQQQSPF
	glutenin	minimal						SQ
		epitope in
		considered
		native form
730	LMW	Glt-156	Immunogenic	Deamidated	DQ2	20	15	QQPPFSEEQESPFS
	glutenin	minimal						Q
		epitope in
		considered
		Deamidated
		form
731	LMW	GLT/GLIA	Immunogenic	Native	DQ2	19	12	QQPPFSQQQQSP
	glutenin	homologue
		peptide 4
732	LMW	Glt-156	Immunogenic	Deamidated	DQ2	20	15	QPPFSEEQESPFSQ
	glutenin	minimal						Q
		epitope
733	LMW	LMW glutenin-	Immunogenic	Native	DQ2	16	14	QPPFSQQQQSPFS
	glutenin	glt-156 (p40-						Q
		p59)
734	LMW	Glt-156	Immunogenic	Native	DQ2	20	15	PPFSQQQQSPFSQ
	glutenin	minimal						QQ
		epitope in
		considered
		native form
735	LMW	Glt-156	Immunogenic	Deamidated	DQ2	20	15	PPFSEEQESPFSQQ
	glutenin	minimal						Q
		epitope in
		considered
		Deamidated
		form
736	LMW	Glt-156	Immunogenic	Native	DQ2	20	15	PFSQQQQSPFSQQ
	glutenin	minimal						QQ
		epitope in
		considered
		native form
737	LMW	Glt-156	Immunogenic	Deamidated	DQ2	20	15	PFSEEQESPFSQQQ
	glutenin	minimal						Q
		epitope in
		considered
		Deamidated
		form
738	LMW	LMW glutenin-	Immunogenic	Native	DQ2	19, 20	10	PFSQQQQSPF
	glutenin	glt-156 (p45-
		p54) minimal
		epitope
739	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	19, 20	10	PFSEQQQSPF
	glutenin	glt-156 (p45-
		p54; E48)
		minimal
		epitope
740	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	20	10	PFSQEQQSPF
	glutenin	glt-156 (p45-
		p54; E49)
		minimal
		epitope
741	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	19, 20	10	PFSQQQESPF
	glutenin	glt-156 (p45-
		p54; E51)
		minimal
		epitope
742	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	19, 20	10	PFSEQQESPF
	glutenin	glt-156 (p45-
		p54; E48 and
		E51) minimal
		epitope
743	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	20	10	PFSEEQQSPF
	glutenin	glt-156 (p45-
		p54; E48 and
		E49) minimal
		epitope
745	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	20	10	PFSQEQESPF
	glutenin	glt-156 (p45-
		p54; E49 and
		E49) minimal
		epitope
746	LMW	LMW glutenin-	Immunogenic	Deamidated	DQ2	20	10	PFSEEQESPF
	glutenin	glt-156 (p45-
		p54; E48, E49
		and E51)
		minimal
		epitope
747	glut-L2	LMW glutenin-	Immunogenic	Native	DQ2.5	19, 90,	9	FSQQQQSPF
		glt-156 (p46-				27
		p54)
748	glut-L2	LMW glutenin-	Immunogenic	Deamidated	DQ2.5	19, 90,	9	FSEQQQSPF
		glt-156 (p46-				27
		p54; E48)
749	glut-L2	LMW glutenin-	Immunogenic	Deamidated	DQ2.5	19, 90,	9	FSQQQESPF
		glt-156 (p46-				27
		p54; E51)
750	glut-L2	LMW glutenin-	Immunogenic	Deamidated	DQ2.5	19, 90,	9	FSEQQESPF
		glt-156 (p46-				27
		p54; E48 and
		E51)
751	LMW	GLT/GLIA	Immunogenic	Native	DQ2	19	15	QQPPFSQQQQPQF
	glutenin	homologue						SQ
		peptide 13
752	LMW	Gluten	Immunogenic	Native	DQ2	61	19	SHQQQPFPQQPYP
	glutenin	peptide #25			(DQ2.5)			QQPYPS
753	gamma-	14-mer-2	Immunogenic	Native	DQ2	57	14	PQQPFPSQQQQPLI
	gliadin	gamma-Glia
		(p173-p186)
754	gamma-	14-mer-2	Immunogenic	Deamidated	DQ2	57	14	PQQPFPSQQEQPLI
	gliadin	gamma-Glia
		(p173-p186)
		in Deamidated
		form
755	LMW	GLT/GLIA	Immunogenic	Native	DQ2	19	15	QQPPFSQQQQPILP
	glutenin	homologue						Q
		peptide 17
756	LMW	Glt-156	Immunogenic	Native	DQ2	21	14	QPPFSQQQQPILPQ
	glutenin	homolog
757	LMW	Glt-156	Immunogenic	Deamidated	DQ2	21	14	QPPFSEQEQPILPQ
	glutenin	homolog in
		Deamidated
		form
762	LMW	GLT/GLIA	Immunogenic	Native	DQ2	19	15	QQPPFSQQQQQPI
	glutenin	homologue						LL
		peptide 16
763	LMW	Glt-156	Immunogenic	Native	DQ2	21	14	QPPFSQQQQQPILL
	glutenin	homolog
764	LMW	Glt-156	Immunogenic	Deamidated	DQ2	21	14	QPPFSEEQEQPILL
	glutenin	homolog in
		Deamidated
		form
765	HMW	Wheat	Immunogenic	Native	DQ2	62	20	QGQQGYYPISPQQS
	glutenin	peptide W21						GQGQQP
		HMW
766	HMW	Wheat	Immunogenic	Native	DQ2	62	16	QGQQGYYPISPQQS
	glutenin	peptide W21						GQ
		HMW
767	HMW	Wheat	Immunogenic	Deamidated	DQ2	62	16	EGQQGYYPISPQQS
	glutenin	peptide W21						GQ
		HMW
768	HMW	Naturally	Immunogenic	Native	DQ8	7	15	GQAGYYPTSPQQS
	glutenin	occurring			(DQ2/8)			GQ
		glutenins
		p722-p736
		(homolog of
		glt04)
769	HMW	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPTSPQQP
	glutenin	occurring			(DQ2/8)			GQ
		glutenins
		p722-p736
		(homolog of
		glt04)
770	HMW	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPISPQQSG
	glutenin	occurring			(DQ2/8)			Q
		glutenins
		p722-p736
		(homolog of
		glt04)
771	HMW	Wheat	Immunogenic	Native	DQ2	62	20	GQGQSGYYPTSPQ
	glutenin	peptide W29						QSGQEAT
772	HMW	Wheat	Immunogenic	Native	DQ2	62	16	GQGQSGYYPTSPQ
	glutenin	peptide W29						QSG
773	HMW	Wheat	Immunogenic	Native	DQ2	62	20	PGQGQSGYYPTSP
	glutenin	peptide W24						QQSGQKQ
		HMW
774	HMW	Wheat	Immunogenic	Native	DQ2	62	16	PGQGQSGYYPTSP
	glutenin	peptide W24						QQS
		HMW
775	HMW	Naturally	Immunogenic	Native	DQ8	7	15	GQSGYYPTSPQQSG
	glutenin	occurring			(DQ2/8)			Q
		glutenins
		(p722-p736)
		(homolog of
		glt04)
776	HMW	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPISPQQLG
	glutenin	occurring			(DQ2/8)			Q
		glutenins
		(p722-p736)
		(homolog of
		glt04)
777	HMW	Naturally	Immunogenic	Native	DQ8	7	15	GQLGYYPTSPQQSG
	glutenin	occurring			(DQ2/8)			Q
		glutenins
		(p722-p736)
		(homolog of
		glt04)
778	HMW	Wheat	Immunogenic	Native	DQ2	62	20	LQPGQGQPGYYPTS
	glutenin	peptide W22						PQQIGQ
		HMW
779	HMW	Wheat	Immunogenic	Native	DQ2	62	16	QGQPGYYPTSPQQI
	glutenin	peptide W22						GQ
		HMW
780	HMW	Naturally	Immunogenic	Native	DQ8	7	15	GQPGYYPTSPQQIG
	glutenin	occurring			(DQ2/8)			Q
		glutenins
		(p722-p736)
		(homolog of
		glt04)
781	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQPGYYPTSPQQP
	Glutenin	occurring			(DQ2/8)			GQ
		glutenins
		(p722-p736)
		( homolog of
		glt04)
782	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQPGYYPTSPQQS
	Glutenin	occurring			(DQ2/8)			GQ
		glutenins
		(p722-p736)
		(homolog of
		glt04)
783	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPTSLQQP
	Glutenin	occurring			(DQ2/8)			GQ
		glutenins
		p722-p736
		(homolog of
		glt04)
784	HMW-	HMW	Immunogenic	Native	DQ8	7	36	SGQGQRPGQWLQ
	Glutenin	glutenin-glt04			(DQ2/8)			PGQGQQGYYPTSP
		(p707-p742)						QQSGQGQQLGQ
785	HMW-	HMW	Immunogenic	Native	DQ8	7	18	PGQGQQGYYPTSP
	Glutenin	glutenin-glt04			(DQ2/8)			QQSGQ
		(p719-p736)
786	HMW-	glt04 (p722-	Immunogenic	Native	DQ8	7	15	GQQGYYPTSPQQS
	Glutenin	p736)			(DQ2/8)			GQ
787	HMW-	glt04 (p722-	Immunogenic	Native	DQ8	7	14	GQQGYYPTSPQQS
	Glutenin	p735)			(DQ2/8)			G
788	HMW-	glt04 (p722-	Immunogenic	Native	DQ8	7	13	GQQGYYPTSPQQS
	Glutenin	p734)			(DQ2/8)
789	HMW-	glt04 (p723-	Immunogenic	Native	DQ8	7	14	QQGYYPTSPQQSG
	Glutenin	p736)			(DQ2/8)			Q
790	HMW-	glt04 (p723-	Immunogenic	Native	DQ8	7	13	QQGYYPTSPQQSG
	Glutenin	p735)			(DQ2/8)
791	HMW-	glt04 (p723-	Immunogenic	Deamidated	DQ8	7, 83	13	QEGYYPTSPQQSG
	Glutenin	p735; E724)			(DQ2/8)
792	HMW-	glt04 (p723-	Immunogenic	Native	DQ8	52	12	QQGYYPTSPQQS
	Glutenin	p734)			(DQ2/8)
793	HMW-	glt04 (p724-	Immunogenic	Native	DQ8	7	12	QGYYPTSPQQSG
	Glutenin	p735)			(DQ2/8)
794	glut-H1	HMW glutenin	Immunogenic	Native	DQ8,	7, 76,	11	QGYYPTSPQQS
		(p724-p734)			DQ8.5	27
797	HMW-	glt04 (p725-	Immunogenic	Native	DQ8	7, 83	11	GYYPTSPQQSG
	Glutenin	p735)			(DQ8.5)
798	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPISPQQP
	Glutenin	occurring			(DQ2/8)			GQ
		glutenins
		(p722-p736)
		(homolog of
		glt04)
799	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPTSPQQS
	Glutenin	occurring			(DQ2/8)			PQ
		glutenins
		(p722-p736)
		(homolog of
		glt04)
800	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPTSPQQL
	Glutenin	occurring			(DQ2/8)			GQ
		glutenins
		(p722-p736)
		(homolog of
		glt04)
801	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPTSPQHP
	Glutenin	occurring			(DQ2/8)			GQ
		glutenins
		(p722-p736)
		(homolog of
		glt04)
802	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQPGYYPTSPLQSG
	Glutenin	occurring			(DQ2/8)			Q
		glutenins
		(p722-p736)
		(homolog of
		glt04)
803	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQHGYYPTSPQLSG
	Glutenin	occurring			(DQ2/8)			Q
		glutenins
		(p722-p736)
		(homolog of
		glt04)
804	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPTSPQQP
	Glutenin	occurring			(DQ2/8)			PQ
		glutenins
		(p722-p736)
		(homolog of
		glt04)
805	HMW-	Naturally	Immunogenic	Native	DQ8	7	15	GQQGYYPTSVQQP
	Glutenin	occurring			(DQ2/8)			GQ
		glutenins
		(p722-p736)
		(homolog of
		glt04)
806	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QPQQPQPFPQQPV
		B04, B17						PQQPQPY
807	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	16	PQQPQPFPQQPVP
		B17						QQP
808	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	PQQPVPQQPQPY
		B04
809	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	PQPFPQQPIPQQP
		B05, B08 in						QPYPQQP
		native form
810	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	20	PQPFPQQPIPEQPQ
		B05, B08 in						PYPQQP
		Deamidated
		form
811	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	16	PQPFPQQPIPQQP
		B05						QPY
812	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	16	PQPFPQQPIPEQPQ
		B05						PY
813	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QQPQPFSQQPIPQ
		B06						QPQPYPQ
814	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	20	QQPQPFSQQPIPEQ
		B06						PQPYPQ
815	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	SQQPIPQQPQPY
		B06
816	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	12	SQQPIPEQPQPY
		B06
817	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QPQPFPQQPIPLQP
		B18						HQPYTQ
818	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	QPQPFPQQPIPL
		B18
819	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	PQPYPQQPQPFPQ
		B13						QPPFCQQ
820	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	16	PQPYPQQPQPFPQ
		B13						QPP
821	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QQPFPQQPFPQQP
		B09, B12, B30						QPYPQQP
822	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	16	QQPFPQQPFPQQP
		B09						QPY
823	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	11	QQPFPQQPFPQ
		B30
824	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	16	PQQPFPQQPQPYP
		B12						QQP
825	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QPQPYPQQPQPYP
		B11						QQPFQPQ
826	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	QPQPYPQQPQPY
		B11
827	gamma-	Glu-21 in	Immunogenic	Native	DQ2	19	21	QPQPFPQQSEQSQ
	gliadin	considered						QPFQPQPF
	or LMW	native form
	glutenin
828	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QPQQPFPQPQQPI
		B03						PYQPQQP
829	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	16	GQQPFPQPEQPIPY
		B03						QG
830	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	QPFPQPQQPIPY
		B03
831	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPIPY
		B03
832	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	WQPQQPFPQPQQ
		B02						PFPLQPQQ
833	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	20	WQPQQPFPQPEQP
		B02						FPLQPQQ
834	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	16	GQQPFPQPEQPFPL
		B02						QG
835	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	QPFPQPQQPFPL
		B02
836	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPFPL
		B02
837	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	LPRPQQPFPWQPQ
		B19						QPFPQPQ
838	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QPQQPFPLQPQQP
		B26						FPWQPQQ
839	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	PFPLQPQQPFPW
		B26
840	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QPQQPFSFSQQPQ
		B29						QPFPLQP
841	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	16	GFSFSQQPEQPFPL
		B29						QG
842	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	FQQPQQSYPVQPQ
		B14						QPFPQPQ
843	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	16	GQSYPVQPEQPFP
		B14						QPG
844	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	SYPVQPQQPFPQ
		B14
845	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	12	SYPVQPEQPFPQ
		B14
846	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	SFSQQPQQPFPL
		B29
847	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	12	SFSQQPEQPFPL
		B29
848	omega-	Wheat	Immunogenic	Deamidated	DQ2	62	16	GQPFPWQPEQPFP
	gliadin	peptide W19						QPG
849	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	YPQQPQPFPQQPIP
		B15						QQPQPY
850	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	QPQPFPQQPIPQ
		B15
851	hor-3	Hor-l	Immunogenic	Native	DQ2.5	88, 62	9	PIPQQPQPY
852	hor-3	Hor-1	Immunogenic	Deamidated	DQ2.5	88, 62	9	PIPEQPQPY
853	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QQQPFPQQPIPQQ
		B16						PQPYPQQ
854	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	11	QQPFPQQPIPQ
		B16
855	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	16	PQQPIPQQPQPYP
		B08						QQP
856	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	16	PQQPIPEQPQPYPQ
		B08						QP
857	Hordein	Barley peptide	Immunogenic	Native	DQ2	62, 86	16	QPQQPIPQQPQPY
		B08						PQQ
858	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62, 86	16	EPEQPIPEQPQPYP
		B08						QQ
859	Hordein	Hordein core	Immunogenic	Native	DQ2	27	9	FPPQQPFPQ
		epitope in
		native form
860	Hordein	Hordein core	Immunogenic	Deamidated	DQ2	27	9	FPPEQPFPQ
		epitope in
		demainated
		form
861	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	PFPQQPQQPFPQP
		B21, B25						QQPFRQQ
862	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	20	PFPQQPQQPFPQP
		B21						EQPFRQQ
863	Hordein	alpha9-	Immunogenic	Native	DQ2	8	14	PQQPFPQPQQPFR
		Hordein in						Q
		native form
864	Hordein	alpha9-	Immunogenic	Deamidated	DQ2	8	14	PQQPFPQPEQPFR
		Hordein in						Q
		Deamidated
		form
865	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	13	QQPFPQPQQPFRQ
		B21
866	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	13	QQPFPQPEQPFRQ
		B21
867	hor-1	Hordein/	Immunogenic	Native	DQ2.5	88, 90,	9	PFPQPQQPF
		Secalin				8, 27,
						62
868	hor-1	Hordein/	Immunogenic	Deamidated	DQ2.5	88, 90,	9	PFPQPEQPF
		Secalin				8, 27,
						62
869	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	PQQPFQPQQPFPQ
		B22						QTIPQQP
870	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	QQPFQPQQPFPQ
		B22
871	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	TFPPSQQPNPLQPQ
		B27						QPFPLQ
872	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	13	PNPLQPQQPFPLQ
		B27
873	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	NPLQPQQPFPLQP
		B23, B24						QPPQQPF
874	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	16	NPLQPQQPFPLQP
		B23						QPP
875	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	16	PLQPQQPFPLQPQP
		B24						PQ
876	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	PQQPQQPFPQPQQ
		B10						PFSWQPQ
877	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	20	PQQPQQPFPQPEQ
		B10						PFSWQPQ
878	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	QPFPQPQQPFSW
		B10
879	Hordein	Barley peptide	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPFSW
		B10
880	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	PQQTIPQQPQQPF
		B28						PLQPQQP
881	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	12	TIPQQPQQPFPL
		B28
882	Hordein	Barley peptide	Immunogenic	Native	DQ2	62	20	QQPFPLQPQQPFP
		B20						QPQPFPQ
883	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	16	GQPFPLQPEQPFPQ
	gliadin	peptide W26						PG
884	gamma-	Wheat	Immunogenic	Deamidated	DQ2	62	12	PFPLQPEQPFPQ
	gliadin	peptide W26
885	gamma-	Barley peptide	Immunogenic	Native	DQ2	62	20	QSQQQFPQPQQPF
	hordein	B07						PQQPQQP
886	gamma-	alpha2-	Immunogenic	Native	DQ2	8	14	QQFPQPQQPFPQQ
	hordein	Hordein in						P
		native form
887	gamma-	alpha2-	Immunogenic	Deamidated	DQ2	8	14	QEFPQPQQPFPQQ
	hordein	Hordein in						P
		Deamidated
		form
888	gamma-	alpha2-	Immunogenic	Deamidated	DQ2	8	14	QQFPQPEQPFPQQ
	hordein	Hordein in						P
		Deamidated
		form
889	gamma-	alpha2-	Immunogenic	Deamidated	DQ2	8	14	QEFPQPEQPFPQQ
	hordein	Hordein in						P
		Deamidated
		form
890	gamma-	Barley peptide	Immunogenic	Native	DQ2	62	12	QQFPQPQQPFPQ
	hordein	B07
891	hor-2	Hordein/	Immunogenic	Native	DQ2.5	90, 8,	9	PQPQQPFPQ
		Secalin				27
892	hor-2	Hordein/	Immunogenic	Deamidated	DQ2.5	90, 8,	9	PQPEQPFPQ
		Secalin				27
893	Secalin	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQPAPIQPEQPFPQ
		R05						QG
894	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	20	PAPIQPQQPFPQQP
		R05, R26						QQPFPQ
895	Secalin	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	PAPIQPEQPFPQ
		R05
896	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	12	PAPIQPQQPFPQ
		R05
897	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	20	FPQQPQQPFPQPQ
		R12						QQLPLQP
898	Secalin	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQQPFPQPEQELPL
		R12						QG
899	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	12	QPFPQPQQQLPL
		R12
900	Secalin	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQELPL
		R12
901	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	20	PTPIQPQQPFPQRP
		R29						QQPFPQ
902	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	12	PFPQRPQQPFPQ
		R29
903	Secalin	gamma1-	Immunogenic	Native	DQ2	90, 27	9	PQQSFPQQP
		Secalin in
		native form
904	Secalin	gamma1-	Immunogenic	Deamidated	DQ2	90, 27	9	PQQSFPEQP
		Secalin in
		Deamidated
		form
905	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	20	FPLQPQQPFPQQPE
		R10						QIISQQ
906	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	12	PFPQQPEQIISQ
		R10
907	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	20	FPQQPEQIISQQPQ
		R25						QPFPLQ
908	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	15	PEQIISQQPQQP
		R25						FPL
909	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	20	PQQLFPLPQQPFPQ
		R22						PQQPFP
910	Secalin	Rye peptide	Immunogenic	Native	DQ2	62	11	LFPLPQQPFPQ
		R22
911	Secalin	alpha9-Secalin	Immunogenic	Native	DQ2	8	14	PQQPFPQPQQPFP
		in native form						Q
912	Secalin	alpha9-Secalin	Immunogenic	Deamidated	DQ2	8	14	PEQPFPQPQQPFP
		in Deamidated						Q
		form
913	Secalin	alpha9-Secalin	Immunogenic	Deamidated	DQ2	8	14	PQQPFPQPEQPFP
		in Deamidated						Q
		form
914	Secalin	alpha9-Secalin	Immunogenic	Deamidated	DQ2	8	14	PEQPFPQPEQPFPQ
		in Deamidated
		form
915	Secalin	alpha2-Secalin	Immunogenic	Native	DQ2	8	14	QPFPQPQQPFPQS
		in native form						Q
916	Secalin	alpha2-Secalin	Immunogenic	Deamidated	DQ2	8	14	QPFPQPEQPFPQS
		in Deamidated						Q
		form
917	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	20	NMQVGPSGQVEW
	secalin	R21						PQQQPLPQ
918	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	16	GMQVGPSGEVEW
	secalin	R21						PQQG
919	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	12	QVGPSGQVEWPQ
	secalin	R21
920	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	12	QVGPSGEVEWPQ
	secalin	R21
921	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	20	SPQPQQPYPQQPF
	secalin	R13, R28						PQQPQQP
922	gamma-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQPEQPYPEQPFP
	secalin	R13						QQG
923	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	12	PQQPYPQQPFPQ
	secalin	R13
924	gamma-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	PEQPYPEQPFPQ
	secalin	R13
925	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	20	PQTQQPQQPFPQP
	secalin	R23						QQPQQLF
926	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	12	PQTQQPQQPFPQ
	secalin	R23
927	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	20	PQEPQQLFPQSQQ
	secalin	R27						PQQPFPQ
928	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	12	PQSQQPQQPFPQ
	secalin	R27
929	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	20	QTQQSIPQPQQPF
	secalin	R17						PQPQQPF
930	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	12	QSIPQPQQPFPQ
	secalin	R17
931	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	20	SIPQPQQPFPQPQQ
	secalin	R02						PFPQSQ
932	gamma-	Rye peptide	Immunogenic	Deamidated	DQ2	62	20	SIPQPQQPFPQPEQ
	secalin	R02						PFPQSQ
933	gamma-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQQPFPQPEQPFP
	secalin	R02						QSG
934	gamma-	Rye peptide	Immunogenic	Deamidated	DQ2	62	13	QPFPQPEQPFPQS
	secalin	R02
935	gamma-	Rye peptide	Immunogenic	Native	DQ2	62	12	QPFPQPQQPFPQ
	secalin	R02
936	gamma-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPFPQ
	secalin	R02
937	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	QYSPYQPQQPFPQ
	Secalin	R07						PQQPTPI
938	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQYSPYQPEQPFPQ
	Secalin	R07						PG
939	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	YSPYQPQQPFPQ
	Secalin	R07
940	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	YSPYQPEQPFPQ
	Secalin	R07
941	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQQPFPQPEQPTPI
	Secalin	R03						QG
942	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	QPFPQPQQPTPIQP
	Secalin	R03, R04						QQPFPQ
943	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	QPFPQPEQPTPI
	Secalin	R03
944	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	QPFPQPQQPTPI
	Secalin	R03
945	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQPTPIQPEQPFPQ
	Secalin	R04						QG
946	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	PTPIQPQQPFPQ
	Secalin	R04
947	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	PTPIQPEQPFPQ
	Secalin	R04
948	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	QQLPLQPQQPFPQ
	Secalin	R01, R09						PQQPIPQ
949	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	QLPLQPQQPFPQ
	Secalin	R09
950	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	QPFPQPQQPIPQ
	Secalin	R01
951	omega-	Sec-gamma1	Immunogenic	Native	DQ2	8	14	PQQPQQSFPQQPQ
	Secalin							R
952	omega-	Sec-gamma1	Immunogenic	Deamidated	DQ2	8	14	PEQPQQSFPQQPQ
	Secalin	in Deamidated						R
		form
953	omega-	Sec-gamma1	Immunogenic	Deamidated	DQ2	8	14	PQQPEQSFPQQPQ
	Secalin	in Deamidated						R
		form
954	omega-	Sec-gamma1	Immunogenic	Deamidated	DQ2	8	14	PQQPQQSFPEQPQ
	Secalin	in Deamidated						R
		form
955	omega-	Sec-gamma1	Immunogenic	Deamidated	DQ2	8	14	PEQPEQSFPQQPQ
	Secalin	in Deamidated						R
		form
956	omega-	Sec-gamma1	Immunogenic	Deamidated	DQ2	8	14	PEQPQQSFPEQPQ
	Secalin	in Deamidated						R
		form
957	omega-	Sec-gamma1	Immunogenic	Deamidated	DQ2	8	14	PQQPEQSFPEQPQ
	Secalin	in Deamidated						R
		form
958	omega-	Sec-gamma1	Immunogenic	Deamidated	DQ2	8	14	PEQPEQSFPEQPQR
	Secalin	in Deamidated
		form
959	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	EQIISQQPFPLQPQ
	Secalin	R20						QPFSQP
960	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	PFPLQPQQPFSQ
	Secalin	R20
961	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQPQQPFPEQPEQI
	Secalin	R6						IG
962	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	PQQPFPQQPEQIIP
	Secalin	R06, R11, R16						QQPQQP
963	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	PQQPFPQQPEQI
	Secalin	R6
964	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	PQQPFPEQPEQI
	Secalin	R6
965	omega-	Rye peptide	Immunogenic	Native	DQ2	62	16	QQPFPQQPEQIIPQ
	Secalin	R11						QP
966	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	EQPFPEQPEQIIPQ
	Secalin	R11						QP
967	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	16	GQPFPQQPEQIIPQ
	Secalin	R11						QG
968	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	PFPQQPEQIIPQ
	Secalin	R11
969	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	PEQIIPQQPQQP
	Secalin	R16
970	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	SQQPQRPQQPFPQ
	Secalin	R08						QPQQIIP
971	omega-	Rye peptide	Immunogenic	Native	DQ2	62	13	RPQQPFPQQPQQI
	Secalin	R08
972	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	QPQQIIPQQPQQPF
	Secalin	R15						PLQPQQ
973	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	IIPQQPQQPFPL
	Secalin	R15
974	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	QQPQQPFPLQPQQ
	Secalin	R14, R19						PVPQQPQ
975	omega-	Rye peptide	Immunogenic	Native	DQ2	62	16	QQPQQPFPLQPQQ
	Secalin	R14						PVP
976	omega-	Rye peptide	Immunogenic	Native	DQ2	62	16	QPFPLQPQQPVPQ
	Secalin	R19						QPQ
977	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	QQPFLLQPQQPFSQ
	Secalin	R18						PQQPFL
978	omega-	Rye peptide	Immunogenic	Native	DQ2	62	11	FLLQPQQPFSQ
	Secalin	R18
979	omega-	Rye peptide	Immunogenic	Native	DQ2	62	20	SPQQPQLPFPQPQ
	Secalin	R24						QPFVVVV
980	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	20	SPQQPQLPFPQPEQ
	Secalin	R24						PFVVVV
981	omega-	Rye peptide	Immunogenic	Native	DQ2	62	12	LPFPQPQQPFVV
	Secalin	R24
982	omega-	Rye peptide	Immunogenic	Deamidated	DQ2	62	12	LPFPQPEQPFVV
	Secalin	R24
983	gamma-	Av-alpha9B in	Immunogenic	Native	DQ2	8	14	QYQPYPEQQQPFV
	avenin	native form						Q
984	gamma-	Av-alpha9B in	Immunogenic	Deamidated	DQ2	8	14	QYQPYPEQEQPFV
	avenin	Deamidated						Q
		form
985	gamma-	Homolog of	Immunogenic	Deamidated	DQ2	62	15	EYQPYPEQEQPILQ
	avenin	oat avenin-						Q
		derived T cell-
		stimulatory
		peptide in
		Deamidated
		form
986	gamma-	Homolog of	Immunogenic	Native	DQ2	62	15	QYQPYPQQQQPIL
	avenin	oat avenin-						QQ
		derived T cell-
		stimulatory
		peptide in
		native form
987	ave-1b	gliadin alpha	Immunogenic	Native	DQ2.5	90, 18,	9	PYPEQQQPF
		avenin-9				8
988	ave-1b	gliadin alpha	Immunogenic	Deamidated	DQ2.5	90, 18,	9	PYPEQEQPF
		avenin-9				8
989	Avenin	T cell	Immunogenic	Native	DQ2	18	31	TTTVQYDPSEQYQP
		recognized						YPEQQEPFVQQQP
		Avenin						PFVQ
		epitope HPLC
		fraction 9
990	Avenin	T cell	Immunogenic	Native	DQ2	18	22	TTTVQYDPSEQYQP
		recognized						YPEQQEPF
		Avenin
		epitope HPLC
		fraction 4
991	Avenin	T cell	Immunogenic	Native	DQ2	18	28	TTTVQYNPSEQYQP
		recognized						YPEQQEPFVQQQP
		Avenin						F
		epitope HPLC
		fraction 9
992	Avenin	T cell	Immunogenic	Native	DQ2	18	27	TTVQYNPSEQYQPY
		recognized						PEQQEPFVQQQPF
		Avenin
		epitope HPLC
		fraction 9
993	Avenin	T cell	Immunogenic	Native	DQ2	18	25	VQYNPSEQYQPYPE
		recognized						QQEPFVQQQPF
		Avenin
		epitope HPLC
		fraction 9
994	Avenin	T cell	Immunogenic	Native	DQ2	18	22	TTTVQYNPSEQYQP
		recognized						YPEQQEPF
		Avenin
		epitope HPLC
		fraction 3
995	Avenin	T cell	Immunogenic	Native	DQ2	18	29	TTTVQYDPSEQYQP
		recognized						YPEQQEPFVQQQQ
		Avenin						PF
		epitope HPLC
		fraction 8
996	Avenin	T cell	Immunogenic	Native	DQ2	18	30	TTTVQYNPSEQYQP
		recognized						YPEQQEPFVQQQQ
		Avenin						PFV
		epitope HPLC
		fraction 8
997	Avenin	T cell	Immunogenic	Native	DQ2	18	29	PSEQYQPYPEQQEP
		recognized						FVQQQQPFVQQQ
		Avenin						QPF
		epitope HPLC
		fraction 9
998	Avenin	Avenin 1490 in	Immunogenic	Native	DQ2	18	19	SEQYQPYPEQQEPF
		native form						VQQQQ
999	Avenin	Avenin 1490 in	Immunogenic	Deamidated	DQ2	18	19	SEQYQPYPEQEEPF
		Deamidated						VQQQQ
		form
1000	Avenin	Av-alpha9A in	Immunogenic	Native	DQ2	8	14	QYQPYPEQQEPFV
		native form						Q
1001	Avenin	Av-alpha9A in	Immunogenic	Deamidated	DQ2	8	14	QYQPYPEQEEPFVQ
		Deamidated
		form
1002	Avenin	Avenin 1505	Immunogenic	Native	DQ2	18	12	YQPYPEQQEPFV
1003	Avenin	Avenin 1504	Immunogenic	Deamidated	DQ2	18	12	YQPYPEQEEPFV
		(deamidated
		form of
		Avenin 1505)
1004	ave-1	gliadin alpha	Immunogenic	Native	DQ2.5	90, 18,	9	PYPEQQEPF
		avenin-9				8
1005	ave-1	gliadin alpha	Immunogenic	Deamidated	DQ2.5	90, 18,	9	PYPEQEEPF
		avenin-9				8
1006	Avenin	Av-gamma2B	Immunogenic	Native	DQ2	8	14	QQPFVQQQQPFVQ
								Q
1007	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVQQQQPFVQ
		in Deamidated						Q
		form
1008	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	QQPFVEQQQPFVQ
		in Deamidated						Q
		form
1009	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVEQQQPFVQ
		in Deamidated						Q
		form
1010	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	QQPFVQEQQPFVQ
		in Deamidated						Q
		form
1011	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVQEQQPFVQ
		in Deamidated
		form
1012	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	QQPFVEEQQPFVQ
		in Deamidated						Q
		form
1013	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVEEQQPFVQ
		in Deamidated						Q
		form
1014	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	QQPFVQQEQPFVQ
		in Deamidated						Q
		form
1015	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVQQEQPFVQ
		in Deamidated						Q
		form
1016	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	QQPFVEQEQPFVQ
		in Deamidated						Q
		form
1017	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVEQEQPFVQ
		in Deamidated						Q
		form
1018	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	QQPFVQEEQPFVQ
		in Deamidated						Q
		form
1019	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVQEEQPFVQ
		in Deamidated						Q
		form
1020	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	QQPFVEEEQPFVQ
		in Deamidated						Q
		form
1021	Avenin	Av-gamma2B	Immunogenic	Deamidated	DQ2	8	14	EQPFVEEEQPFVQQ
		in Deamidated
		form
1022	Avenin	Avenin core	Immunogenic	Native	DQ2	27	9	FVQQQQQPF
		epitope in
		native form
1023	Avenin	Avenin core	Immunogenic	Deamidated	DQ2	27	9	FVQQQEQPF
		epitope in
		Deamidated
		form
1024	gamma-	Glu-21	Immunogenic	Native	DQ2	19	12	QSEQSQQPFQPQ
	gliadin	minimal
	or LMW	epitope in
	glutenin	considered
		native form
1025	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5 or	90, 17,	9	EQPQQPYPQ
	gamma 3,				DQ8	23
	glia-
	gamma
	1b
1026	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5 or	90, 17,	9	QQPQQPYPE
	gamma 3,				DQ8	23
	glia-
	gamma
	1b
1027	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5 or	90, 78,	9	EQPEQPYPQ
	gamma 3,				DQ8	23
	glia-
	gamma
	1b
1028	glia-	CAUTION,	Immunogenic	Deamidated	DQ2.5 or	90, 78,	9	QQPEQPYPE
	gamma 3,	100% match			DQ8	23
	glia-	to Candida
	gamma	protein/
	1b	gamma gliadin
1029	glia-	gamma 5	Immunogenic	Deamidated	DQ2.5	90, 78,	9	SQPEQQFPQ
	gamma 3	gliadin				23
1030	glia-	gamma 5	Immunogenic	Deamidated	DQ2.5	90, 78,	9	SQPQQEFPQ
	gamma 3	gliadin				23
1031	glia-	gammaVII-	Immunogenic	Deamidated	DQ2.5	90, 17,	9	QQPQQPFPE
	gamma	gliadin				25
	1b, glia-
	gamma 4c
1032	glia-	gammaVII-	Immunogenic	Deamidated	DQ2.5	90, 17,	9	QQPEQPFPE
	gamma	gliadin				25, 8
	4c, glia-
	gamma
	1b
1033	glia-	gammaVII-	Immunogenic	Deamidated	DQ2.5	90, 17,	9	EQPEQPFPE
	gamma	gliadin				25
	4c, glia-
	gamma
	1b
1034	glia-	gamma gliadin	Immunogenic	Native	DQ2.5	90, 62,	9	PQPQQPFCQ
	gamma 4c					78, 81
1035	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5	90, 62,	9	PQPEQPFCQ
	gamma					78, 81
	4d
1036	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5	90, 62,	9	PQPQQPFCE
	gamma					78, 81
	4d
1037	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5	90, 62,	9	PQPEQPFCE
	gamma					78, 81
	4d
1038	glia-	gliadin omega	Immunogenic	Native	DQ2.5	90, 86	9	PQPQQPFSQ
	gamma	1
	4e
1039	glia-	gliadin omega	Immunogenic	Deamidated	DQ2.5	90, 86	9	PQPEQPFSQ
	gamma	1
	4e
1040	glia-	gliadin omega	Immunogenic	Native	DQ2.5	86, 90	9	PFPQPQQPI
	omega 3	3
1041	glia-	gliadin omega	Immunogenic	Deamidated	DQ2.5	86, 90	9	PFPQPEQPI
	omega 3	3
1042	glia-	CAUTION	Immunogenic	Native	DQ2.5	86, 90	9	PQPQQPIPV
	omega 4	100% match
		to 2 Prunus sp.
		peptides/
		gliadin omega
		4
1043	glia-	gliadin omega	Immunogenic	Deamidated	DQ2.5	86, 90	9	PQPEQPIPV
	omega 4	4
1044	glia-	gliadin omega	Immunogenic	Native	DQ2.5	86, 90	9	LQPQQPFPQ
	omega 5	5
1045	glia-	gliadin omega	Immunogenic	Deamidated	DQ2.5	86, 90	9	LQPEQPFPQ
	omega 5	5
1046	Avenin Q	avenin-gliadin	Immunogenic	Native	DQ2 or	87	14	QQPFMQQQQPFM
		like			DQ8			QP
1047	Avenin Q-	avenin-gliadin	Immunogenic	Native	DQ2 or	88	14	QQPFVQQQQQPFV
	5	like			DQ8			Q
1048	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5	90, 74	9	PEQSFPQQQ
	gamma 1	1
1049	glia-	gamma gliadin	Immunogenic	Deamidated	DQ2.5	90, 74	9	PEQSFPQQE
	gamma 1
1050	ave-1 06	Avenin gliadin	Immunogenic	Native	DQ2.5	88	16	QYQPYPEQQQPILQ
		like						QQ
1051	ave-1 06	avenin-gliadin	Immunogenic	Deamidated	DQ2.5	88	16	QYQPYPEQEQPILQ
		like						QQ
1052	ave-1 04	avenin-gliadin	Immunogenic	Native	DQ2.5	88	16	QQYQPYPQQQPF
		like						MQPL
1053	ave-1 04	avenin-gliadin	Immunogenic	Deamidated	DQ2.5	88	16	EQYQPYPEQQPFM
		like						QPL
CD1D multimer and ligand:

In one embodiment, the peptide is a human, non-human, or synthetic/engineered peptide. For example, the peptide is a Minor Histocompatibility Antigen (MiHA).

Exemplary MiHAs are known in the art, e.g., in Spierings et al. Tissue Antigens 2014 84:347-360; which is incorporated by reference herein. MiHAs are typically utilized in embodiments relating to transplantation, where it refers to epitopes that are created because of protein sequence variation in polymorphic proteins.

Residues of the antigenic peptide that engages the MHC binding groove can loosely be divided into two types: anchor residues, which engage with the MHC molecule and confer stability to the MHC-peptide complex (for example, see residues P2, P3, P5, P6, P7, and P9 in Bowness et al. 1999 Expert Reviews in Molecular Medicine 16:1-10; which is incorporated by reference herein in its entirety), and interfacial residues, which are solvent-exposed and can engage with the cognate T-cell receptor (see, e.g., residues P1, P4, and P8 in Bowness). A featureless peptide is a peptide in which anchor residues are preserved, while interfacial residues of the peptide are mutated to alanine or glycine residues to prevent TCR binding A featureless peptide-MHC is therefore a MHC peptide complex in which the presented peptide is a featureless peptide. Featureless peptide MHCs are typically used in embodiments relating to transplantation tolerance. In patients receiving MHC-mismatched solid organ or hematopoietic stem cell transplants, use of a CAL T cell presenting the relevant donor-mismatched featureless peptide-MHC CAL can permit selective depletion of recipient alloreactive T cells targeted towards this mismatched donor HLA allele.

The MHC can be a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In embodiments where more than one MHC unit is present, e.g., the MHC is oligomeric, the units can be provided in series, e.g., in a chain, or provided arrayed in one or more dimensions around a central point or linker, provided conjugated/bound in any geometry to a scaffold molecule, or any combination of the foregoing. Naturally occurring examples of TCR recognition domain structures in the art include MHC dimers (Lebowitz et al., 1999 Cellular Immunology 192:175-184), tetramers (Altman et al., 1996 Science 274:94-96), pentamers (proimmune.com/introduction-to-pentamers/), octamers (Guillame et al., 2003 JBC 278:4500-4509), dextramers (Batard et al., 2006 Journal of Immunological Methods 310:136-148), dodecamers (Huang et al., 2016 PNAS 113:E1890-7), lipid vesicles (Mallet-Designe et al., 2003 The Journal of Immunology 170:123-131), and quantum dots (Chattopadhyay et al., 2006 Nature Medicine 12:972-7). Each of the foregoing references is incorporated by reference herein in its entirety.

In some embodiments, the TCR recognition domain can comprise a CD1 domain (e.g., a CD1d domain), e.g., a sequence comprising an extracellular domain of CD1, (e.g., CD1). As used herein, “cluster of differentiation 1 family member d” or “CD1d” refers to a cell surface protein that displays lipid antigens to T cells. The sequences of several CD1d isoforms, and the structure of CD1d are known in the art, see, e.g., the 3 isoforms provided in the NCBI database for CD1d (Gene ID 912), and Bagchi et al., 2018 and Oleinika et al., Nature Communcations 2018 9:684; which are incorporated by reference herein in their entireties. For example, isoform 1 of CD1d is SEQ ID NO: 5 (NCBI Ref Seq NP 001757.1), isoform 2 of CD1d is SEQ ID NO: 6 (NCBI Ref Seq NP 001306074.1), and isoform 3 of CD1d is SEQ ID NO: 7 (NCBI Ref Seq NP_001358690.1). In some embodiments, the CD1d domain comprises a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of SEQ ID NOs: 5-7. In some embodiments, the CD1d domain comprises a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to the extracellular domain of one of SEQ ID NOs: 5-7 (e.g., amino acids 20-301 of SEQ ID NO: 5). In some embodiments, the CD1d domain comprises a sequence with at least at least 95% sequence identity to one of SEQ ID NOs: 5-7 and retains the lipid binding activity of the wild-type reference sequence. In some embodiments, the CD1d domain comprises a sequence with at least 95% sequence identity to the extracellular domain of one of SEQ ID NOs: 5-7 (e.g., amino acids 20-301 of SEQ ID NO: 5) and retains the lipid binding activity of the wild-type reference sequence.

In some embodiments, the CD1 domain further comprises a ligand, e.g., a non-peptide ligand. Exemplary, non-limiting ligands of CD1 domains include but are not limited to:

Protein (Allele) Ligand

	human CD1a	C-glycoside
		GSL-1
		OCH
	human CD1b	C-glycoside
		GSL-1
		OCH
	human CD1c	C-glycoside
		GSL-1
		OCH
	human CD1d	OCH
	human CD1d	PBS-57

In some embodiments, a TCR recognition domain can comprise sequences or molecules in addition to the, e.g., MHC, pMHC, of CD1 domains and sequences. For example, it can further comprise other polypeptide and/or non-polypeptide components that enable multimerization. Exemplary components that permit multimerization can include biotin (non-polypeptide) and/or streptavidin polypeptide that is used to permit tetramerization.

In various embodiments, protein interaction domains are found on an extracellular portion of the respective polypeptides.

As used herein, “recognition polypeptide” refers to an extracellular polypeptide having a ligand-binding domain. In some embodiments, the ligand-binding domain can be an antibody reagent. In some embodiments, the recognition polypeptide can further comprise a protein interaction domain.

As used herein, “signaling polypeptide” refers to a transmembrane polypeptide having an intracellular signaling domain, e.g., a T cell receptor (TCR) signaling domain. In some embodiments, the signaling polypeptide can further comprise a protein interaction domain. In some embodiments, the signaling polypeptide can further comprise an extracellular protein interaction domain.

As used herein, “biomolecular interaction domain” refers to a domain that permits specific binding of two separate molecules to each other. The molecules can be or can comprise polypeptides. In some embodiments, one or both of the molecules or biomolecular interaction domains can be a non-peptide. When a pair of biomolecular interaction domains is provided herein, they permit two or more molecules to bind specifically, e.g. one of the biomolecular interaction domains can bind specifically to the second biomolecular interaction domain. In some embodiments, specific binding can occur when two separate biomolecular interaction domains, e.g., of a pair, are present. In some embodiments, specific binding can occur when three or more separate biomolecular interaction domains are present. It is noted that protein interaction domains are a type of biomolecular interaction domains and where one is specified herein, the other may always be substituted.

As used herein, when a molecule is referred to as a protein or polypeptide, it comprises a protein, peptide, or polypeptide sequence, but may comprise additional motifs, modifications, or domains of a non-proteinaceous nature. A number of exemplary biomolecular interaction domains, as well as pairs of protein interaction domains are provided elsewhere herein. In some embodiments, the biomolecular interaction domains comprise, consist, or consist essentially of proteins or polypeptides. In some embodiments, the biomolecular interaction domains comprise, consist, or consist essentially of non-proteinaceous molecules. In some embodiments, one of a pair of biomolecular interaction domains can comprise, consist, or consist essentially of proteins or polypeptides and the second of the pair of biomolecular interactions domains can comprise, consist, or consist essentially of a non-proteinaceous molecule (e.g., FITC and anti-FITC). Exemplary protein interaction domains are known in the art and can be used in embodiments of the aspects described herein.

As used herein, “protein interaction domain” refers to a domain that permits specific binding of two separate polypeptides to each other. A number of exemplary protein interaction domains, as well as pairs of protein interaction domains are provided elsewhere herein. In some embodiments, the protein interaction domains of the polypeptides of a multi-component CAL and/or CAR can bind specifically, e.g. one of the protein interaction domains can bind specifically to a second protein interaction domain of the multi-component CAL and/or CAR. In some embodiments, specific binding can occur when two separate protein interaction domains are present. In some embodiments, specific binding can occur when three or more separate protein interaction domains are present. Exemplary protein interaction domains are known in the art and can be used in embodiments of the aspects described herein.

In some embodiments of any of the aspects described herein, the protein interaction domains can be leucine zipper domains. Leucine zipper domains are a type of protein-protein interaction domain commonly found in transcription factors characterized by leucine residues evenly spaced through a α-helix. Leucine zippers may form heterodimers or homodimers. A number of leucine zipper domains, as well as their ability to bind each other, are known in the art and discussed further, e.g., in Reinke et al. JACS 2010 132:6025-31 and Thompson et al. ACS Synth Biol 2012 1:118-129; each of which is incorporated by reference herein in its entirety. In some embodiments, one leucine zipper domain is BZip (RR) and the second leucine zipper domain is AZip (EE). In some embodiments, the sequence of a BZip (RR) leucine zipper domain is MDPDLEIRAAFLRQRNTALRTEVAELEQEVQRLENEVSQYETRYGPLGGGK (SEQ ID NO: 3). In some embodiments, the sequence of a AZip (EE) leucine zipper domain is MDPDLEIEAAFLERENTALETRVAELRQRVQRLRNRVSQYRTRYGPLGGGK (SEQ ID NO: 4).

Further exemplary leucine zipper domains are described in Reinke et al. JACS 2010 132:6025-31; which is incorporated by reference herein in its entirety. For example, suitable leucine zipper domains can include SYNZIP 1 to SYNZIP 48, and BATF, FOS, ATF4, ATF3, BACH1, JUND, NFE2L3, and HEPTAD. Binding affinities of various combinations of these domains are described, e.g., at FIG. 1 of Reinke et al. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 1000 nM or less. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 100 nM or less. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 10 nM or less. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 1 nM or less.

Further exemplary pairs of protein interaction domains can include a) PSD95-Dlg1-zo-1 (PDZ) domains; b) a streptavidin domain and a streptavidin binding protein (SBP) domain; and c) a PYL domain and an ABI domain.

In some embodiments of any of the aspects described herein, the protein interaction domains can be chemically-induced protein interaction domains, e.g., domains that will only bind specifically in the presence of a third molecule, e.g., a small molecule or drug. Exemplary pairs of chemically-induced protein interaction domains can include: FKBP-binding domain of mTOR (FRB) and FK506 binding protein (FKBP) (binding of which is activated by tacrolimus, everolimus, or a rapalog); cyclophilin-Fas fusion protein (CyP-Fas) and FK506 binding protein (FKBP) (binding of which is activated by FKCsA); calcineurin A (CNA) and FK506 binding protein (FKBP) (binding of which is activated by FK506); gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) (binding of which is activated by gibberellin); Snap-tag and Halo tag (binding of which is activated by HaXS); and T14-3-3-cdeltaC and C-Terminal peptides of PMA2 (CT52) (binding of which is activated by fusicoccin). Further description of chemically-induced protein interaction domains can be found in the art, e.g., Miyamoto et al. Nat Chem Biol. 2012 Mar. 25; 8(5): 465-470 and Belshaw et al. PNAS 1996 93:4604-4607; each of which is incorporated herein by reference in its entirety.

In some embodiments of any of the aspects described herein, the protein interaction domains can comprise at least one nucleotide tag and at least one zinc finger domain. Zinc finger domains are characterized by the coordination of a zinc ion in order to stabilize their tertiary structure. The particular folds that appear in zinc fingers can vary. In some embodiments, a zinc finger domain can be a nucleotide-binding zinc finger domain. In some embodiments, a zinc finger domain can be a DNA-binding zinc finger domain. In some embodiments, the protein interaction domain of the recognition polypeptide is a nucleotide tag and the extracellular protein interaction domain of the signaling polypeptide is a zinc finger domain. In some embodiments, a nucleotide tag can be a DNA tag. In some embodiments, a nucleotide tag can be a dsDNA tag comprising the entire recognition sequence for the zinc finger domain being used. Exemplary zinc finger domains and their cognate nucleotide tags are described in the art, e.g. Mali et al. Nature Methods 2013 10:403-406; which is incorporated by reference herein in its entirety. In some embodiments, a zinc finger domain can be sZF15 as described in Mali et al. Nature Methods 2013 10:403-406.

In one embodiment, the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE).

In some embodiments of any of the aspects described herein, the protein interaction domains can comprise a pair of substantially complementary nucleotide tags, e.g., fully complementary or complementary enough to hybridize specifically. The degree of complementarity necessary may vary depending on the total length of the tags and G/C content of the complementary portions. One of skill in the art can readily determine the relevant affinity necessary for tags of a given size and G/C content. In some embodiments, a nucleotide tag can be a DNA tag.

In some embodiments, a nucleotide tag can be a DNA tag. In some embodiments, a nucleotide tag can be a dsDNA tag.

In one embodiment, the protein interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of protein interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1).

In one embodiment, the protein interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of protein interaction domains are collectively a Snap-tag and a Halo tag.

In one embodiment, the protein interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of protein interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52).

In one embodiment, the protein interaction domain(s) is a PYL and/or a ABI, or any binding pair of protein interaction domains are collectively a PYL and a ABI.

In one embodiment, the biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding protein or any binding pair of biomolecular interaction domains are collectively a FITC and a FITC binding protein.

In one embodiment, the biomolecular interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of biomolecular interaction domains are collectively a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein.

In some embodiments, the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain (i.e. a CAL) binds specifically to a native cell surface molecule on a NK cell, dendritic cell, or T cell. In such embodiments, a subject can be treated as described herein without administration of an engineered cell. In some embodiments, the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain (i.e. a CAL) binds specifically to a native cell surface molecule on a NK cell. In some embodiments, the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain (i.e. a CAL) binds specifically to a native cell surface molecule on dendritic cell. Exemplary native cell surface molecules for such embodiments include, but are not limited to CD3. Other suitable cell surface molecules are the CD cell surface proteins. CDs are known in the art and one of ordinary skill can readily select one expressed by the desired cell type(s). For example, see the HCDM database at hcdm.org and the lists available on the world wide web at chemeurope.com/en/encyclopedia/List_of_human_clusters_of_differentiation.html; and docs.abcam.com/pdf/immunology/Guide-to-human-CD-antigens.pdf. Accordingly, exemplary biomolecular interaction domains for such embodiments include but are not limited to an anti-CD3 antibody reagent, or a Fab domain. One of skill in the art is aware of other cell surface molecules found, or found exclusively, on the cell surface of the relevant cell type(s), as well as multiple reagents that can bind (e.g., bind specifically) to such cell surface molecules.

In aspects with a single recognition polypeptide and a single signaling polypeptide that are able to bind specifically without a third polypeptide, the multiple-component CALs or CARs described herein will activate in the presence of the target ligand, thereby inducing T cell activity in the vicinity of the target ligand. Further described herein are multiple-component CALs or CARs capable of logic computation, for example, multiple-component CALs or CARs that serve as AND, OR, or NOT logic gates.

In some aspects, described herein are compositions that comprise components of a multi-component CAL and/or CAR that permits AND gate logic. In these aspects, activation of the multi-component CAL and/or CAR happens only in the presence of two target ligands; recognition of a single target ligand is not sufficient for activation. Such multi-component CALs or CARs can permit greater specificity and reduce off-target effects. Any single ligand that is a good marker for a target cell or tissue may occur elsewhere in a subject, resulting in off-target effects. However, requiring the recognition of two separate marker ligands reduces the odds of off-target activity.

In one embodiment, a nucleotide tag can be a DNA tag or dsDNA tag.

Further embodiments of AND logic gate multi-component CALs and CARs are described herein.

In some embodiments of any of the aspects described herein, the compositions comprise components of a multi-component CAL and/or CAR that are NOT logic gate. For example, recognition of a second target ligand by a second recognition polypeptide can prevent interaction (e.g. specific binding) of the signaling polypeptide and first recognition polypeptide. Such embodiments can permit suppression of T cell activity in inappropriate and/or off-target tissues. For example, the second target ligand can be a marker of a tissue that is particularly sensitive to T cell activity, is a known area of off-target activity, and/or shares markers with the desired target tissue. In some embodiments, in a NOT gate multi-component CAL and/or CAR, the second target ligand is not a ligand found in the target tissue and/or cells, e.g., in or on a disease T cell. In some embodiments, the second target ligand of a NOT logic gate multi-component CAL and/or CAR is found on a healthy and/or non-target cell and not on a diseased and/or target cell. Various 2- and 3-dimensional configurations of such pairs of nucleotide pairs are known in the art.

In some embodiments, the target ligand recognized by the second recognition polypeptide is found on a healthy and/or non-target cell and not on a diseased and/or target cell. In some embodiments, the protein interaction domain of the second recognition polypeptide and the protein interaction domain of the first recognition polypeptide have a greater affinity than the protein interaction domain of the signaling polypeptide and the protein interaction domain of the first recognition polypeptide. In some embodiments, the protein interaction domain of the second recognition polypeptide and the protein interaction domain of the signaling polypeptide have a greater affinity than the protein interaction domain of the signaling polypeptide and the protein interaction domain of the first recognition polypeptide. Relative binding affinities can be determined experimentally, e.g., by binding affinity assays known in the art and relative binding affinities are known for a number of combinations of protein interaction domains described herein, see, e.g. Reinke et al. JACS 2010 132:6025-31; which is incorporated by reference herein in its entirety. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domains can be at least 2x greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domains can be at least 5x greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domains can be at least 10x greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain.

As used herein, “target ligand” refers to a molecule in or on a cell which can be bound by a ligand-binding domain. Non-limiting examples of such molecules can include polypeptides, lipids, saccharides, and the like. In some embodiments, the target ligand can be an extracellular molecule. In some embodiments, the target ligand can be a cell surface molecule.

In some embodiments, e.g., those relating to a multi-component CAL and/or CAR with a single recognition polypeptide or an AND gate multi-component CAL and/or CAR, the target ligand (e.g. the first and/or second target ligand) can be a ligand expressed in a target tissue. In some embodiments, the target ligand can be expressed constitutively in the target tissue and/or cell. In some embodiments, the target ligand can be expressed exclusively in the target tissue and/or cell. In some embodiments, the target ligand can be expressed at a higher level in the target tissue and/or cell than in other tissues and/or cells. As recognition of a target ligand in embodiments relating to a multi-component CAL and/or CAR with a single recognition polypeptide or an AND gate multi-component CAL and/or CAR can result in T cell activation (e.g. cell killing activity of the cell comprising the target ligand), the target ligand can be selected to target T cell activity in a desirable and/or therapeutic way, e.g., by targeting a disease cell. In some embodiments, a target ligand is a ligand found in/on a diseased and/or target cell. In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is a ligand found in/on a diseased and/or target cell. In some embodiments, a target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is a ligand found on a diseased and/or target cell and not on a healthy and/or non-target cell. In some embodiments, the diseased cell is an autoreactive or alloreactive T cell. In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is found on the surface of a disease cell. In some embodiments, a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR specifically binds to a target ligand on the surface of a disease cell, e.g. as compared to binding to normal cells.

In some embodiments, a composition and/or cell described herein can further comprise a second multi-component CAL and/or CAR according to any of the aspects and embodiment described herein for the first multi-component CAL and/or CAR. By way of non-limiting example, a second CAL and/or CAR can be designed to bind specifically to (and, e.g., be activated by or inhibited by) different target ligands than those to which the first multi-component CAL and/or CAR specifically binds (and, e.g. is activated by or inhibited by). This can provide increased specificity, reduced off-target effects, and/or reduced effective dosages for the methods described herein. In some embodiments, the recognition domain of second multi-component CAL and/or CAR bind specifically to different target ligands than those bound by the recognition domain of the first multi-component CAL and/or CAR. In some embodiments, the antibody reagents of second multi-component CAL and/or CAR bind specifically to different target ligands than those bound by the antibody reagents of the first multi-component CAL and/or CAR.

In some embodiments, the second multi-component CAL and/or CAR can comprise an inhibitory intracellular signaling domain, e.g., T cell receptor (TCR) signaling domain, e.g., one that inhibits engineered cell, e.g., T cell, activity. In such embodiments, the second multi-component can therefore be designed to operate in opposition to the first multi-component CAL and/or CAR, e.g. permitting inhibition of T cell activation while the first multi-component CAL and/or CAR permits activation of T cell activity. Inhibitory intracellular TCR signaling domains are known in the art and can include, by way of non-limiting example, PD1; CTLA4; BTLA; KIR; LAG-3; TIM-3; A2aR; LAIR-1; and TGIT. In one embodiment, non-active mimetics of an activating TCR can be used.

In some embodiments, described herein is a composition comprising a cell, a CAL, and a CAR (e.g., with the CAR comprising either an inhibitory domain or costimulation/activation domain).

In one embodiment, the TCR recognition domain comprises a MHC allogeneic to the cell. In one embodiment, the TCR recognition domain comprises a peptide allogeneic to the cell. In one embodiment, the TCR recognition domain comprises a MHC allogeneic to the target cell. In one embodiment, the TCR recognition domain comprises a peptide allogeneic to the target cell. In one embodiment, the TCR recognition domain comprises a non-self peptide relative to the target cell.

In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with the signaling polypeptide of the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain is a ligand found on a healthy and/or non-target cell. In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with the signaling polypeptide of the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain is a ligand found on a healthy and/or non-target cell and not on a diseased and/or target cell. In some embodiments, the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain can be an OR logic gate according to any of the embodiments described herein and the second target ligand can be a ligand found in/on, or specific to, diseased cells.

In some embodiments of any of the aspects, a ligand-binding domain can comprise or consist essentially of an antibody reagent. In some embodiments, the antibody reagent can be an immunoglobulin molecule, a monoclonal antibody, a chimeric antibody, a CDR-grafted antibody, a human antibody, a humanized antibody, a Fab, a Fab′, a F(ab′)2, a Fv, a disulfide linked Fv, a scFv, a single domain antibody, a diabody, a multispecific antibody, a dual specific antibody, an anti-idiotypic antibody, and/or a bispecific antibody.

In some embodiments, the intracellular signaling domain can be a T-cell activation domain. In some embodiments, the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of: TCRζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD3ζ, CD22, CD79a, CD79b, CD66d, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD223 (LAG3), CD270 (HVEM), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), DAP10, LAT, NKD2C SLP76, TRIM, and ZAP70. In some embodiments, the signaling domain can be a paralog or ortholog of any of the foregoing.

Multi-Component CAR/CAL Cells

Presented herein are cells that express the compositions or multi-component CARs and/or CALs presented herein. A cell can be any cell, for example, any mammalian cell, e.g., a human cell. In one embodiment, the cell is a dendritic cell, regulatory T cell, or effector T cell. In one embodiment the cell is a dendritic cell (CAL DC), a T cell (e.g., effector, regulatory, etc.) (CAL-T); regulatory T cell, effector T cell, natural killer cell (CAL NK), or any other myeloid cell.

In one aspect, described herein is an engineered cell expressing and/or comprising one or more multi-component CARs/CALs, or a composition comprising the same as described herein, e.g., at least one signaling polypeptide and at least one recognition polypeptide. In some embodiments, the cell is a natural killer (NK) cell, dendritic cell, regulatory T cell, effector T cell. Such cells expressing and/or comprising both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAR/CAL are referred to herein as “complete multi-component CAR/CAL” cells. In some embodiments, a complete multi-component CAR/CAL cell expresses both a signaling polypeptide (e.g., a CAR) and at least one recognition polypeptide (e.g. a CAL or adaptor as described elsewhere herein) of a multi-component CAL and/or CAR. In some embodiments, a complete multi-component CAL and/or CAR cell comprises nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR. In some embodiments, a signaling polypeptide is present on the membrane of a cell. In some embodiments, the one or more recognition polypeptides are present in the extracellular space, e.g., the recognition polypeptide(s) can be expressed and secreted by the cell or the cell can be contacted by recognition polypeptides provided from another source (e.g. produced synthetically or by another cell and optionally, purified or processed before the contacting step).

In any of the aspects described herein, e.g., those relating to either a complete or partial multi-component CAL and/or CAR cell, the recognition and/or signaling polypeptide can be under the control of an inducible and/or repressible promoter. Such promoters allow the expression of the polypeptide to be increased or decreased as desired and are in contrast to constitutive promoters. The term “constitutively active promoter” refers to a promoter of a gene which is expressed at all times within a given cell. Exemplary promoters for use in mammalian cells include cytomegalovirus (CMV), Elongation Factor 1a (EF1a), and the like. The term “inducible promoter” refers to a promoter of a gene which can be expressed in response to a given signal, for example addition or reduction of an agent. Non-limiting examples of an inducible promoter are promoters that are regulated in a specific tissue type, a promoter regulated by a steroid hormone, by a polypeptide hormone (e.g., by means of a signal transduction pathway), or by a heterologous polypeptide (e.g., the tetracycline-inducible systems, “Tet-On” and “Tet-Off”; see, e.g., Clontech Inc., CA, Gossen and Bujard, Proc. Natl. Acad. Sci. USA 89:5547, 1992, and Paillard, Human Gene Therapy 9:983, 1989; each of which are incorporated by reference herein in its entirety). In some embodiments, expression of the polypeptide can be precisely regulated, for example, by using an inducible regulatory sequence that is sensitive to certain physiological regulators, e.g., circulating glucose levels, or hormones (Docherty et al., 1994, FASEB J. 8:20-24). Such inducible expression systems, suitable for the control of expression in cells or in mammals include, for example, regulation by ecdysone, by estrogen, progesterone, tetracycline, chemical inducers of dimerization, and isopropyl-beta-D1-thiogalactopyranoside (IPTG). A person skilled in the art would be able to choose the appropriate regulatory/promoter sequence based on the intended use of the polypeptide.

In some embodiments, the expression of one or more of the recognition or signaling polypeptides can be constitutive. In some embodiments, the expression of one or more of the recognition or signaling polypeptides can be transient. Transient expression can be achieved by, e.g., use of transient and/or inducible expression promoters or by use of transient vectors, e.g. those that do not incorporate into the genome and/or persist in the target cell. By way of non-limiting example, derivatives of viruses such as the bovine papillomavirus (BPV-1), or Epstein-Barr virus (pHEBo, pREP-derived and p205) can be used for transient expression of nucleic acids in eukaryotic cells. For other suitable expression systems as well as general recombinant procedures, see Molecular Cloning A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press: 1989) Chapters 16 and 17; which is incorporated by reference herein in its entirety. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the recognition polypeptide can be transiently expressed. In some embodiments, the recognition polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the signaling polypeptide can be transiently expressed. In some embodiments, the recognition polypeptide of a multi-component CAL can be constitutively expressed and the signaling polypeptide can be provided exogenously. In some embodiments, the recognition polypeptide of a multi-component CAL can be transiently expressed and the signaling polypeptide can be provided exogenously. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the recognition polypeptide can be provided exogenously. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be transiently expressed and the recognition polypeptide can be provided exogenously.

The CARs and CALs described herein can be produced according to any method known in the art, e.g., recombinant expression or peptide synthesis. Exemplary methods can include, the NIH Tetramer Core Facility's MHC expression protocols (available on the world wide web at tetramer.yerkes.emory.edu/support/protocols #1); ProImmune's pentamer protocols (available on the world wide web at proimmune.com/protocols-2/); Immudex's dextramer protocols (available on the word wide web at immudex.com/resources/protocols/) and the CAR T cell production protocols provided in the “Primary Human T cells Isolation and Culture” and “Lentiviral Transduction of Human T cells” sub-sections in “Method Details” section of Cho et al., 2018 Cell 173:1426-1438; each of which is incorporated by reference herein in its entirety.

In one aspect, described herein is a method of killing a target cell, the method comprising contacting the cell with a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein. In some embodiments, the target cell can be a diseased cell, e.g., an autoreactive or alloreactive T cell. In one aspect, described herein is a method of treating or preventing a disease, e.g., an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection, or GvHD, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein. In one aspect, described herein is a method of treating or preventing autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein.

Another aspect provided herein is a method of preventing and/or treating a malignant T cell condition in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein. In one aspect, described herein is a method of treating or preventing a malignant T cell condition in a subject, comprising administering a complete multi-component CAR/CAL, CAR and/or CAL as according to any of the embodiments described herein. In one aspect, described herein is a method of treating or preventing a malignant T cell condition in a subject, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein

In some embodiments, the complete multi-component CAL and/or CAR cell can be autologous or allogeneic to the subject. In some embodiments, the complete multi-component CAL and/or CAR cell can be derived and/or descended from a cell obtained from the subject or a third party and has been modified ex vivo to comprise the at least one multi-component CAL and/or CAR, e.g., genetically engineered to comprise nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR. In some embodiments, the method can further comprise the steps of obtaining a cell from a subject (e.g. a dendritic cell, regulatory T cell, or effector T cell), altering the cell to comprise nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR, and then administering the cell to the subject.

In one embodiment, the engineered cell is further modified to lack or have reduced expression of the native MHCI/II, e.g., as measured on the cell surface. Methods for engineering a cell to reduce or eliminate the native MHCI/II are known in the art and include, e.g., expression of RNA interference (such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.), expression of an inhibitory oligonucleotide, nuclease-based inhibition (such as CRISPR, TALEN, Meganuclease, etc.), and expression of a KDEL-motif (SEQ ID NO: 2749) containing binding protein capable of restricting MHCI/II to the endoplasmic reticulum. One skilled in the art can assess whether knockdown of native MHCI/II is achieved by assessing mRNA or protein levels of MHCI/II via, e.g., PCR-based assays or western blotting, respectively.

In one embodiment, the cell is further engineered to knockout the native MHCI/II. Methods for engineering a cell to knockout the native MHCI/II are known in the art and include, e.g., expression of RNA interference (such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.), expression of an inhibitory oligonucleotide, nuclease-based inhibition (such as CRISPR, TALEN, Meganuclease, etc.). One skilled in the art can assess whether knockout of native MHCI/II is achieved by assessing mRNA or protein levels of MHCI/II via, e.g., PCR-based assays or western blotting, respectively.

In one aspect, described herein is an engineered cell expressing and/or comprising one or more of the compositions according to any of the embodiments described herein. In one aspect, described herein is an engineered cell expressing and/or comprising one or more multi-component CAL and/or CAR signaling polypeptides according to any of the embodiments described herein. In some embodiments, the cell is a dendritic cell, regulatory T cell, or effector T cell. In some embodiments, the cell is a T cell. Such cells expressing and/or comprising a multi-component CAL and/or CAR signaling polypeptide are referred to herein as “partial multi-component CAL” cells or “partial multi-component CAR” cells. In some embodiments, the partial multi-component CAL and/or CAR cell does not express, e.g., does not comprise a nucleic acid sequence encoding, a multi-component CAL and/or CAR recognition polypeptide. In some embodiments, a partial multi-component CAL and/or CAR cell comprises a nucleic acid sequence encoding at least one multi-component CAL and/or CAR signaling polypeptide. In some embodiments, the multi-component CAL and/or CAR signaling polypeptide is present on the membrane of the cell, e.g., is expressed as a transmembrane protein at detectable levels. In some embodiments, the signaling polypeptide further comprises a secondary protein interaction domain that specifically binds with the protein interaction domain of the second recognition polypeptide, e.g., the signaling polypeptide is part of an AND gate multi-component CAL and/or CAR as described elsewhere herein. In some embodiments, the cell can further comprise a second multi-component CAL and/or CAR signaling polypeptide, e.g., a signaling polypeptide that is part of a second multi-component CAL and/or CAR according to any of the embodiments described herein.

In one aspect, described herein is a method of killing a target cell, the method comprising contacting the target cell with a partial multi-component CAL and/or CAR cell according to any of the embodiments described herein and contacting the target cell with at least one recognition polypeptide of the multi-component CAL and/or CAR. In some embodiments, the target cell can be a diseased cell, e.g., an autoreactive or alloreactive T cell. In some embodiments, the target cell can be a diseased cell, e.g., a cancer cell (e.g., a T cell- or T cell precursor cell-derived neoplasm, hereafter referred to as “T cell neoplasms”. In some embodiments, the method can further comprise the steps of obtaining a cell from a subject (e.g. a NK cell, a dendritic cell, regulatory T cell, or effector T cell), altering the cell to comprise a nucleic acid sequence encoding a signaling polypeptide of a multi-component CAL and/or CAR, and then administering the cell to the subject.

In some embodiments of any of the methods described herein, a pair of protein interaction domains of a multi-component CAL and/or CAR can comprise chemically induced binding domains and the method can further comprise administering a compound that induces binding of the domains. In some embodiments, when one protein interaction domain is FKBP-binding domain of mTOR (FRB) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering tacrolimus, a rapalog, or everolimus. In some embodiments, when one protein interaction domain is cyclophilin-Fas fusion protein (CyP-Fas) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering FKCsA. In some embodiments, when one protein interaction domain is calcineurin A (CNA) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering FK506. In some embodiments, when one protein interaction domain is gibberellin insensitive (GIA) and a second protein interaction domain is gibberellin insensitive dwarf1 (GID1), the method further comprises administering gibberellin. In some embodiments, when one protein interaction domain is Snap-tag and a second protein interaction domain is Halo tag, the method further comprises administering HaXS. In some embodiments, when one protein interaction domain is T14-3-3-cdeltaC and a second protein interaction domain is C-Terminal peptides of PMA2 (CT52), the method further comprises administering fusicoccin.

In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be engineered. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be transgenic. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be recombinant. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a T cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a human T cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a T cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a human T cell.

It is specifically contemplated herein that each of the individual embodiments described herein can be combined, e.g., in a single cell. By way of non-limiting example, a single cell could comprise a first complete multi-component CAL and/or CAR and a second partial multi-component CAL and/or CAR, wherein each multi-component CAL and/or CAR can be according to any of the embodiments described herein.

In some embodiments, the methods described herein relate to CAL-immune cell therapies. In some embodiments, the methods described herein relate to CAR-immune cell therapies such as CAR-T therapy. Standard CAR-T and related therapies relate to adoptive cell transfer of immune cells (e.g. T cells) expressing a CAR that binds specifically to a targeted cell type (e.g. disease cells, e.g., autoreactive or alloreactive T cells) to treat a subject, e.g., for an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD.

In some embodiments, the cells administered as part of the therapy can be autologous to the subject. In some embodiments, the cells administered as part of the therapy are not autologous to the subject. In some embodiments, the cells are engineered and/or genetically modified to express a multi-component CAL and/or CAR, or portion thereof as described herein. Further discussion of CAR-T therapies can be found, e.g., in Maus et al. Blood 2014 123:2624-35; Reardon et al. Neuro-Oncology 2014 16:1441-1458; Hoyos et al. Haematologica 2012 97:1622; Byrd et al. J Clin Oncol 2014 32:3039-47; Maher et al. Cancer Res 2009 69:4559-4562; and Tamada et al. Clin Cancer Res 2012 18:6436-6445; each of which is incorporated by reference herein in its entirety.

In some embodiments, the technology described herein relates to a syringe or catheter, including an organ-specific catheter (e.g., renal catheter, biliary catheter, cardiac catheter, etc.), comprising a therapeutically effective amount of a composition described herein.

Methods of Treatment

In some embodiments, the methods described herein relate to the treatment or prevention of transplant rejection in a subject having a cell, tissue, or organ transplant with one or more compositions, CALs, CARs, or cells as described herein. In another embodiment, the methods described herein relate to the treatment or prevention of GvHD in a subject having a cell, tissue, or organ transplant with one or more compositions, CALs, CARs, or cells as described herein.

As used herein, “GvHD” refers to a disease characterized by the active process of donor cells attacking the recipient's own cells. GvHD can develop soon after a transplant, e.g., within weeks or months (acute GvHD), or can occur much later after the transplant, e.g., at least 3-6 months later (chronic GvHD). Symptoms of acute GvHD include, but are not limited to, skin rash or blisters, abdominal pain or discomfort, diarrhea, jaundice, and edema. Symptoms of chronic GvHD include, but are not limited to, changes to skin or nail texture, hair loss or thinning, muscle pain or weakness, blurred vision, mouth sores, shortness of breath, persistent cough, abdominal pain or discomfort, and diarrhea.

A subject can be identified as having or be at risk of having GvHD by a skilled clinician. Diagnostic tests useful in identifying a subject having GvHD are known in the art and will vary based on the type of transplant the subject has received. The diagnosis of GvHD is made by, for example, physical examination for the signs and symptoms for GvHD known in the art, serologic testing for dysfunction of the liver, gall bladder, kidney, and hematopoietic cells, histologic analysis of biopsies obtained from affected organs, and radiologic imaging of affected organs. In one embodiment, the method further comprises administering at least a second therapeutic. In one embodiment, the composition, CARs, CALs, or cells described herein are administered in combination with Abatacept (Orencia®) or Belatacept (Nulojix®). Abatacept and Belatacept, developed by Bristol-Meyers Squibb, are fusion proteins composed of the Fc region of the immunoglobulin IgG1 fused to the extracellular domain of CTLA-4. Abatacept is currently approved by the FDA for treatment of rheumatoid arthritis. Belatacept, which only differs from Abatacept by two amino acids, is an immunosuppressant intended to prevent rejection following a kidney transplant.

In one embodiment, the transplant is vascularized composite allograft (VCA). In one embodiment, the transplant is any human or non-human cell, tissue, or organ. In another embodiment, the transplant is any type of transplants procedures, e.g., any heart transplant, any lung transplant, any liver transplant, any pancreas transplant, any cornea transplant, any trachea transplant, any kidney transplant, any skin transplant, any pancreatic islet cell transplant, any allograft (e.g., a transplantation of allogeneic tissue), any xenograft (e.g., a transplantation of xenogeneic tissue), or any autograft (e.g., a transplantation of tissue). A skilled practitioner will be able to perform a transplant or identify a subject having had a transplant using standard procedural protocols.

In some embodiments, the methods described herein relate to the treatment or prevention of an autoimmune diseases or conditions, or hypersensitivity reaction I-IV, or immune reaction against foreign therapeutic proteins/molecules, or T cell mediated inflammation or immune response; with compositions, CARs, CALs, or cells as described herein. Subjects having an autoimmune disease can be identified by a physician using current methods of diagnosing an autoimmune disease. Symptoms and/or complications of an autoimmune disease which characterize these conditions and aid in diagnosis are well known in the art and include but are not limited to, fatigue, achy muscles, swelling and redness, low-grade fever, numbness or tingling of the hands or feet, hair loss, and/or skin rash. Tests that may aid in a diagnosis of, e.g. autoimmune disease include, but are not limited to, blood counts, and an antinuclear antibody test (ANA). A family history of autoimmune disease, or having risk factors for autoimmune disease (e.g. gender, age, ethnicity, and exposure to environmental agents, such as procainamide, hydralazine, mercury, gold, or silver) can also aid in determining if a subject is likely to have autoimmune disease or in making a diagnosis of autoimmune disease.

As used herein, the term “autoimmune disease”, “autoimmune condition”, or “autoimmune disease or disorder” herein is a disease or disorder arising from and directed against an individual's own tissues or a co-segregate or manifestation thereof or resulting condition therefrom.

Auto-immune related diseases and disorders arise from an overactive and/or abnormal immune response of the body against substances (autoantigens) and tissues normally present in the body, otherwise known as self or autologous substance. This dysregulated inflammatory reaction causes an exaggerated response by macrophages, granulocytes, lymphocytes, and/or T-lymphocytes leading to abnormal tissue damage and cell death. Subsequent loss of function is associated with inflammatory tissue damage.

Autoantigens, as used herein, are endogenous proteins or fragments thereof that are involved in or elicit this pathogenic immune response. Autoantigen can be any substance, or a portion thereof normally found within a mammal that, in an autoimmune disease, becomes the primary (or a primary, or secondary) target of attack by the immune system. The term also includes antigenic substances that induce conditions having the characteristics of an autoimmune disease when administered to mammals. Additionally, the term includes peptidic subclasses consisting essentially of immunodominant epitopes or immunodominant epitope regions of autoantigens. Immunodominant epitopes or regions in induced autoimmune conditions are fragments of an autoantigen that can be used instead of the entire autoantigen to induce the disease. In humans afflicted with an autoimmune disease, immunodominant epitopes or regions are fragments of antigens specific to the tissue or organ under autoimmune attack and recognized by a substantial percentage (e.g. a majority though not necessarily an absolute majority) of autoimmune attack T-cells.

Autoantigens that are known to be associated with autoimmune disease include myelin proteins with demyelinating diseases, e.g. multiple sclerosis and experimental autoimmune myelitis; collagens and rheumatoid arthritis; insulin, proinsulin, glutamic acid decarboxylase 65 (GAD65); and islet cell antigen (ICA512; ICA12) for insulin dependent diabetes.

A common feature in a number of autoimmune related diseases and inflammatory conditions is the involvement of pro-inflammatory CD4+ T cells. These T cells are responsible for the release of inflammatory, Th1 type cytokines. Cytokines characterized as Th1 type include interleukin 2 (IL-2), γ-interferon, TNFα and IL-12. In some embodiments, cytokines characterized as Th1 type include interleukin 2 (IL-2), interferon γ, and TNFα. Such pro-inflammatory cytokines act to stimulate the immune response, in many cases resulting in the destruction of autologous tissue. Cytokines associated with suppression of T cell response are the Th2 type, and include IL-10, IL-4 and TGF-β. It has been found that Th1 and Th2 type T cells may use the identical antigen receptor in response to an immunogen; in the former producing a stimulatory response and, in the latter, a suppressive response.

T cell mediated inflammation, or a T cell mediated immune response is inflammation and/or an immune response in which T cells and/or T cell activity contributes to or originates the inflammation/immune response.

As used herein, “inflammation” refers to the complex biological response to harmful stimuli, such as pathogens, damaged cells, or irritants. Inflammation is a protective attempt by the organism to remove the injurious stimuli as well as initiate the healing process for the tissue. Accordingly, the term “inflammation” includes any cellular process that leads to the production of pro-inflammatory cytokines, inflammation mediators and/or the related downstream cellular events resulting from the actions of the cytokines thus produced, for example, fever, fluid accumulation, swelling, abscess formation, and cell death. Inflammation can include both acute responses (i.e., responses in which the inflammatory processes are active) and chronic responses (i.e., responses marked by slow progression and formation of new connective tissue). Acute and chronic inflammation may be distinguished by the cell types involved. Acute inflammation often involves polymorphonuclear neutrophils; whereas chronic inflammation is normally characterized by a lymphohistiocytic and/or granulomatous response. An inflammatory condition is any disease state characterized by inflammatory tissues (for example, infiltrates of leukocytes such as lymphocytes, neutrophils, macrophages, eosinophils, mast cells, basophils and dendritic cells) or inflammatory processes which provoke or contribute to the abnormal clinical and histological characteristics of the disease state.

As used herein, an “immune response” refers to a response by a cell of the immune system, such as a B cell, T cell (CD4 or CD8), regulatory T cell, antigen-presenting cell, dendritic cell, monocyte, macrophage, NKT cell, NK cell, basophil, eosinophil, or neutrophil, to a stimulus (e.g., to an a disease, an antigen, or healthy cells, e.g., in the case of autoimmunity). In some embodiments of the aspects described herein, an immune response is a T cell response, such as a CD4+ response or a CD8+ response. Such responses by these cells can include, for example, cytotoxicity, proliferation, cytokine or chemokine production, trafficking, or phagocytosis, and can be dependent on the nature of the immune cell undergoing the response. Stimulation of an immune response refers to an induction or increase of the immune response. Suppression of an immune response refers to an elimination or decrease of the immune response.

A “cell-mediated immune response” is one mediated by T-cells and/or other white blood cells. A “cell-mediated immune response” is elicited by the presentation of antigenic epitopes in association with Class I or Class II molecules of the major histocompatibility complex (MHC), CD1 or other non-classical MHC-like molecules. This activates antigen-specific CD4+ T helper cells or CD8+ cytotoxic lymphocyte cells (“CTLs”). CTLs have specificity for peptide antigens that are presented in association with proteins encoded by classical or non-classical MHCs and expressed on the surfaces of cells. CTLs help induce and promote the intracellular destruction of intracellular microbes, or the lysis of cells infected with such microbes. Another aspect of cellular immunity involves an antigen-specific response by helper T-cells. Helper T-cells act to help stimulate the function, and focus the activity of, nonspecific effector cells against cells displaying peptide or other antigens in association with classical or non-classical MHC molecules on their surface. A “cell-mediated immune response” also refers to the production of cytokines, chemokines and other such molecules produced by activated T-cells and/or other white blood cells, including those derived from CD4+ and CD8+ T-cells. The stimulation of a cell-mediated immunological response may be determined by a number of assays, such as by lymphoproliferation (lymphocyte activation) assays, CTL cytotoxic cell assays, by assaying for T-lymphocytes specific for the antigen in a sensitized subject, or by measurement of cytokine production by T cells in response to re-stimulation with antigen. Such assays are well known in the art. See, e.g., Erickson et al. (1993) J. Immunol. 151:4189-4199; and Doe et al. (1994) Eur. J. Immunol. 24:2369-2376.

In some embodiments, the T cell mediated immune response is a response to a drug administered to the subject. It is contemplated herein that the present technology can be utilized for depletion of anti-drug specific T cells to prevent immune responses against administered biologics, cell therapies, and/or gene therapies. For example, the methods and compositions described herein can be used to prevent or treat anti-AAV and anti-transgene immune responses for administered adeno-associated virus (AAV) gene therapies, preventing or treating immune to responses to genome editing agents such as CRISPR/Cas9, Transcription activator-like effector nucleases (TALENs), or Zinc Finger Nucleases (ZFNs), and preventing or treating immune responses to enzyme replacement therapies such as recombinant human acid α-glucosidase (Pompe disease), a-L-iduronidase (Mucopolysaccharidosis I), and α-galactosidase (Fabry disease).

In one embodiment of any one of the methods described, the autoimmune disorder is selected from the group consisting of thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis), palindromic arthritis, inflammatory hyperproliferative skin diseases, psoriasis such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and psoriasis of the nails, atopy including atopic diseases such as hay fever and Job's syndrome, dermatitis including contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, non-specific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic intraocular inflammatory diseases, urticaria such as chronic allergic urticaria and chronic idiopathic urticaria, including chronic autoimmune urticaria, myositis, polymyositis/dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic scleroderma), sclerosis such as systemic sclerosis, multiple sclerosis (MS) such as spino-optical MS, primary progressive MS (PPMS), and relapsing remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, sclerosis disseminata, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (for example, Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis such as ulcerative colitis, colitis ulcerosa, microscopic colitis, collagenous colitis, colitis polyposa, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), bowel inflammation, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome, including adult or acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, an autoimmune hematological disorder, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as in meningitis, herpes gestationis, pemphigoid gestationis, pruritis scroti, autoimmune premature ovarian failure, sudden hearing loss due to an autoimmune condition, IgE-mediated diseases such as anaphylaxis and allergic and atopic rhinitis, encephalitis such as Rasmussen's encephalitis and limbic and/or brainstem encephalitis, uveitis, such as anterior uveitis, acute anterior uveitis, granulomatous uveitis, nongranulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome such as chronic or acute glomerulonephritis such as primary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membrano- or membranous proliferative GN (MPGN), including Type I and Type II, and rapidly progressive GN, proliferative nephritis, autoimmune polyglandular endocrine failure, balanitis including balanitis circumscripta plasmacellularis, balanoposthitis, erythema annulare centrifugum, erythema dyschromicum perstans, eythema multiform, granuloma annulare, lichen nitidus, lichen sclerosus et atrophicus, lichen simplex chronicus, lichen spinulosus, lichen planus, lamellar ichthyosis, epidermolytic hyperkeratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, allergic reaction, eczema including allergic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and vesicular palmoplantar eczema, asthma such as asthma bronchiale, bronchial asthma, and auto-immune asthma, conditions involving infiltration of T cells and chronic inflammatory responses, immune reactions against foreign antigens such as fetal A-B-O blood groups during pregnancy, chronic pulmonary inflammatory disease, autoimmune myocarditis, leukocyte adhesion deficiency, lupus, including lupus nephritis, lupus cerebritis, pediatric lupus, non-renal lupus, extra-renal lupus, discoid lupus and discoid lupus erythematosus, alopecia lupus, systemic lupus erythematosus (SLE) such as cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and lupus erythematosus disseminatus, juvenile onset (Type I) diabetes mellitus, including pediatric insulin-dependent diabetes mellitus (IDDM), adult onset diabetes mellitus (Type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic large-artery disorder, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, sarcoidosis, granulomatosis including lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitides, including vasculitis, large-vessel vasculitis (including polymyalgia rheumatica and giant-cell (Takayasu's) arteritis), medium-vessel vasculitis (including Kawasaki's disease and polyarteritis nodosa/periarteritis nodosa), microscopic polyarteritis, immunovasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, necrotizing vasculitis such as systemic necrotizing vasculitis, and ANCA-associated vasculitis, such as Churg-Strauss vasculitis or syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs positive anemia, Diamond Blackfan anemia, hemolytic anemia or immune hemolytic anemia including autoimmune hemolytic anemia (AIHA), pernicious anemia (anemia perniciosa), Addison's disease, pure red cell anemia or aplasia (PRCA), Factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases involving leukocyte diapedesis, CNS inflammatory disorders, multiple organ injury syndrome such as those secondary to septicemia, trauma or hemorrhage, antigen-antibody complex-mediated diseases, anti-glomerular basement membrane disease, anti-phospholipid antibody syndrome, allergic neuritis, Behcet's disease/syndrome, Castleman's syndrome, Goodpasture's syndrome, Reynaud's syndrome, Sjogren's syndrome, Stevens-Johnson syndrome, pemphigoid such as pemphigoid bullous and skin pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, pemphigus mucus-membrane pemphigoid, and pemphigus erythematosus), autoimmune polyendocrinopathies, Reiter's disease or syndrome, an immune complex disorder such as immune complex nephritis, antibody-mediated nephritis, polyneuropathies, chronic neuropathy such as IgM polyneuropathies or IgM-mediated neuropathy, and autoimmune or immune-mediated thrombocytopenia such as idiopathic thrombocytopenic purpura (ITP) including chronic or acute ITP, scleritis such as idiopathic cerato-scleritis, episcleritis, autoimmune disease of the testis and ovary including autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis such as autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis), or subacute thyroiditis, idiopathic hypothyroidism, Grave's disease, polyglandular syndromes such as autoimmune polyglandular syndromes (or polyglandular endocrinopathy syndromes), paraneoplastic syndromes, including neurologic paraneoplastic syndromes such as Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis such as allergic encephalomyelitis or encephalomyelitis allergica and experimental allergic encephalomyelitis (EAE), myasthenia gravis such as thymoma-associated myasthenia gravis, cerebellar degeneration, neuromyotonia, opsoclonus or opsoclonus myoclonus syndrome (OMS), and sensory neuropathy, multifocal motor neuropathy, Sheehan's syndrome, autoimmune hepatitis, lupoid hepatitis, giant-cell hepatitis, autoimmune chronic active hepatitis, lymphoid interstitial pneumonitis (LIP), bronchiolitis obliterans (non-transplant) vs NSIP, Guillain-Barre syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis such as primary biliary cirrhosis and pneumonocirrhosis, autoimmune enteropathy syndrome, Celiac or Coeliac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, amylotrophic lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear disease such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsed or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome/nonsyphilitic interstitial keratitis, Bell's palsy, Sweet's disease/syndrome, rosacea autoimmune, zoster-associated pain, amyloidosis, a non-cancerous lymphocytosis, a primary lymphocytosis, which includes monoclonal B cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathy, paraneoplastic syndrome, channelopathies including channelopathies of the CNS, autism, inflammatory myopathy, focal or segmental or focal segmental glomerulosclerosis (FSGS), endocrine opthalmopathy, uveoretinitis, chorioretinitis, autoimmune hepatological disorder, fibromyalgia, multiple endocrine failure, Schmidt's syndrome, adrenalitis, gastric atrophy, presenile dementia, demyelinating diseases such as autoimmune demyelinating diseases and chronic inflammatory demyelinating polyneuropathy, Dressler's syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility, e.g., due to anti-spermatozoan antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortion, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, bird-fancier's lung, allergic granulomatous angiitis, benign lymphocytic angitis, Alport's syndrome, alveolitis such as allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reaction, Sampter's syndrome, Caplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial lung fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum et diutinum, erythroblastosis fetalis, eosinophilic faciitis, Shulman's syndrome, Felty's syndrome, cyclitis such as chronic cyclitis, heterochronic cyclitis, iridocyclitis (acute or chronic), or Fuch's cyclitis, Henoch-Schonlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndromes, Evan's syndrome, autoimmune gonadal failure, Sydenham's chorea, post-streptococcal nephritis, thromboangitis ubiterans, thyrotoxicosis, tabes dorsalis, chorioiditis, giant-cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplant organ reperfusion, retinal autoimmunity, aphthae, aphthous stomatitis, arteriosclerotic disorders, aspermiogenesis, autoimmune hemolysis, Boeck's disease, enteritis allergica, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, febris rheumatica, Hamman-Rich's disease, sensoneural hearing loss, ileitis regionalis, leucopenia, transverse myelitis, primary idiopathic myxedema, ophthalmia symphatica, polyradiculitis acuta, pyoderma gangrenosum, acquired spenic atrophy, vitiligo, toxic-shock syndrome, conditions involving infiltration of T cells, leukocyte-adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, diseases involving leukocyte diapedesis, multiple organ injury syndrome, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathies, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine failure, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid, frontal, maxillary, or sphenoid sinusitis, an eosinophil-related disorder such as eosinophilia, pulmonary infiltration eosinophilia, eosinophilia-myalgia syndrome, Loffler's syndrome, chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, granulomas containing eosinophils, seronegative spondyloarthritides, polyendocrine autoimmune disease, sclerosing cholangitis, sclera, episclera, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia telangiectasia syndrome, angiectasis, autoimmune disorders associated with collagen disease, rheumatism, allergic hypersensitivity disorders, glomerulonephritides, reperfusion injury, ischemic re-perfusion disorder, lymphomatous tracheobronchitis, inflammatory dermatoses, dermatoses with acute inflammatory components, and autoimmune uveoretinitis (AUR). In some embodiments, the autoimmune disease or condition or T cell mediated inflammation can be neurodegeneration, e.g, Alzheimer's or Parkinson disease.

In some embodiments of any of the aspects, the autoimmune disease or condition, or T cell mediated inflammation can be type 1 diabetes, rheumatoid arthritis, multiple sclerosis, pemphigus, alopecia, lupus, vitiligo, or chronic fatigue syndrome.

As used herein, “malignant T cell condition” refers to a condition in which T cells display one or more of uncontrolled growth (i.e., division beyond normal limits), invasion (i.e., intrusion on and destruction of adjacent tissues), and metastasis (i.e., spread to other locations in the body via lymph or blood). Non-limiting examples of malignant T cell conditions include T cell cancers, lymphoma, leukemia, T cell acute lymphoblastic leukemia, and T cell lymphoblastic lymphoma.

The compositions and methods described herein can be administered to a subject to treat or prevent an autoimmune diseases or conditions; T cell mediated inflammation or immune response; or transplant rejection. In some embodiments, the methods described herein comprise administering an effective amount of compositions, CALs, CARs, or cells described herein to a subject in order to alleviate a symptom of an autoimmune diseases or conditions; T cell mediated inflammation or immune response; or transplant rejection. As used herein, “alleviating a symptom” is ameliorating any condition or symptom associated with the disease or condition. As compared with an equivalent untreated control, such reduction is by at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, 99% or more as measured by any standard technique. A variety of means for administering the compositions described herein to subjects are known to those of skill in the art. Such methods can include, but are not limited to oral, parenteral, intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, cutaneous, topical, injection, or intratumoral administration. Administration can be local or systemic.

The administration of the compositions contemplated herein may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. In a preferred embodiment, compositions are administered parenterally. The phrases “parenteral administration” and “administered parenterally” as used herein refers to modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravascular, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intratumoral, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. In one embodiment, the compositions contemplated herein are administered to a subject by direct injection into a tumor, lymph node, or site of infection.

It can generally be stated that a pharmaceutical composition comprising the cells, e.g., T cells or CAL cells or CAR cells, described herein may be administered at a dosage of 10² to 10¹⁰ cells/kg body weight, preferably 10⁵ to 10⁶ cells/kg body weight, including all integer values within those ranges. The number of cells will depend upon the ultimate use for which the composition is intended as will the type of cells included therein. For uses provided herein, the cells are generally in a volume of a liter or less, can be 500 mLs or less, even 250 mLs or 100 mLs or less. Hence the density of the desired cells is typically greater than 10⁶ cells/ml and generally is greater than 10⁷ cells/ml, generally 10⁸ cells/ml or greater. The clinically relevant number of immune cells can be apportioned into multiple infusions that cumulatively equal or exceed 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, or 10¹² cells. In some aspects of the present invention, particularly since all the infused cells will be redirected to a particular or specific target antigen, lower numbers of cells, in the range of 10⁶/kilogram (10⁶-10¹¹ per patient) may be administered. CAL and/or CAR expressing cell compositions may be administered multiple times at dosages within these ranges. The cells may be allogeneic, syngeneic, xenogeneic, or autologous to the patient undergoing therapy. If desired, the treatment may also include administration of mitogens (e.g., PHA) or lymphokines, cytokines, and/or chemokines (e.g., IFN-γ, IL-2, IL-12, TNF-alpha, IL-18, and TNF-beta, GM-CSF, IL-4, IL-13, Flt3-L, RANTES, MIP1α, etc.) as described herein to enhance induction of the immune response.

In some embodiments, the dosage can be from about 1×10⁵ cells to about 1×10⁸ cells per kg of body weight. In some embodiments, the dosage can be from about 1×10⁶ cells to about 1×10⁷ cells per kg of body weight. In some embodiments, the dosage can be about 1×10⁶ cells per kg of body weight. In some embodiments, one dose of cells can be administered. In some embodiments, the dose of cells can be repeated, e.g., once, twice, or more. In some embodiments, the dose of cells can be administered on, e.g., a daily, weekly, or monthly basis.

The dosage ranges for the agent, e.g., a CAL, CAR, cell, or composition described herein depend upon the potency, and encompass amounts large enough to produce the desired effect e.g., prevention of transplant rejection, reduction in inflammation, etc. The dosage should not be so large as to cause unacceptable adverse side effects. Generally, the dosage will vary with the age, condition, and sex of the patient and can be determined by one of skill in the art. The dosage can also be adjusted by the individual physician in the event of any complication. In some embodiments, the dosage ranges from 0.001 mg/kg body weight to 0.5 mg/kg body weight. In some embodiments, the dose range is from 5 μg/kg body weight to 100 μg/kg body weight. Alternatively, the dose range can be titrated to maintain serum levels between 1 μg/mL and 1000 μg/mL. For systemic administration, subjects can be administered a therapeutic amount, such as, e.g., 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, or more.

Administration of the doses recited above can be repeated. In some embodiments, the doses are given once a day, or multiple times a day, for example but not limited to three times a day. In some embodiments, the doses recited above are administered daily for several weeks or months. The duration of treatment depends upon the subject's clinical progress and responsiveness to therapy.

In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg. In some embodiments, the dose can be about 2 mg/kg. In some embodiments, the dose can be about 4 mg/kg. In some embodiments, the dose can be about 5 mg/kg. In some embodiments, the dose can be about 6 mg/kg. In some embodiments, the dose can be about 8 mg/kg. In some embodiments, the dose can be about 10 mg/kg. In some embodiments, the dose can be about 15 mg/kg. In some embodiments, the dose can be from about 100 mg/m² to about 700 mg/m². In some embodiments, the dose can be about 250 mg/m². In some embodiments, the dose can be about 375 mg/m². In some embodiments, the dose can be about 400 mg/m². In some embodiments, the dose can be about 500 mg/m².

In some embodiments, the dose can be administered intravenously. In some embodiments, the intravenous administration can be an infusion occurring over a period of from about 10 minutes to about 3 hours. In some embodiments, the intravenous administration can be an infusion occurring over a period of from about 30 minutes to about 90 minutes.

In some embodiments the dose can be administered about weekly. In some embodiments, the dose can be administered weekly. In some embodiments, the dose can be administered weekly for from about 12 weeks to about 18 weeks. In some embodiments the dose can be administered about every 2 weeks. In some embodiments the dose can be administered about every 3 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered about every 2 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered about every 3 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered intravenously about every 2 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered intravenously about every 3 weeks. In some embodiments, the dose can be from about 200 mg/m² to about 400 mg/m² administered intravenously about every week. In some embodiments, the dose can be from about 200 mg/m² to about 400 mg/m² administered intravenously about every 2 weeks. In some embodiments, the dose can be from about 200 mg/m² to about 400 mg/m² administered intravenously about every 3 weeks. In some embodiments, a total of from about 2 to about 10 doses are administered. In some embodiments, a total of 4 doses are administered. In some embodiments, a total of 5 doses are administered. In some embodiments, a total of 6 doses are administered. In some embodiments, a total of 7 doses are administered. In some embodiments, a total of 8 doses are administered. In some embodiments, the administration occurs for a total of from about 4 weeks to about 12 weeks. In some embodiments, the administration occurs for a total of about 6 weeks. In some embodiments, the administration occurs for a total of about 8 weeks. In some embodiments, the administration occurs for a total of about 12 weeks. In some embodiments, the initial dose can be from about 1.5 to about 2.5 fold greater than subsequent doses.

In some embodiments, the dose can be from about 1 mg to about 2000 mg. In some embodiments, the dose can be about 3 mg. In some embodiments, the dose can be about 10 mg. In some embodiments, the dose can be about 30 mg. In some embodiments, the dose can be about 1000 mg. In some embodiments, the dose can be about 2000 mg. In some embodiments, the dose can be about 3 mg given by intravenous infusion daily. In some embodiments, the dose can be about 10 mg given by intravenous infusion daily. In some embodiments, the dose can be about 30 mg given by intravenous infusion three times per week.

A therapeutically effective amount is an amount of an agent that is sufficient to produce a statistically significant, measurable change in, or prevent the occurrence of an autoimmune disease or condition; T cell mediated inflammation or immune response; transplant rejection; or GvHD. Such effective amounts can be gauged in clinical trials as well as animal studies.

An agent can be administered intravenously by injection or by gradual infusion over time. Given an appropriate formulation for a given route, for example, agents useful in the methods and compositions described herein can be administered intravenously, intranasally, by inhalation, intraperitoneally, intramuscularly, subcutaneously, intracavity, and can be delivered by peristaltic means, if desired, or by other means known by those skilled in the art. It is preferred that the compounds used herein are administered orally, intravenously or intramuscularly. Local administration, e.g., directly to the site of an organ or tissue transplant is also specifically contemplated.

Therapeutic compositions containing at least one agent can be conventionally administered in a unit dose, for example. The term “unit dose” when used in reference to a therapeutic composition refers to physically discrete units suitable as unitary dosage for the subject, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required physiologically acceptable diluent, i.e., carrier, or vehicle.

The compositions are administered in a manner compatible with the dosage formulation, and in a therapeutically effective amount. The quantity to be administered and timing depends on the subject to be treated, capacity of the subject's system to utilize the active ingredient, and degree of therapeutic effect desired.

In embodiments where the subject is administered a partial multi-component CAL and/or CAR cell and a recognition polypeptide, the partial multi-component CAL and/or CAR cell and a recognition polypeptide can be administered together or separately. In embodiments where the subject is separately administered a partial multi-component CAL and/or CAR cell and a recognition polypeptide each of the compositions can be administered, separately, according to any of the dosages and administration routes/routines described herein.

Precise amounts of active ingredient required to be administered depend on the judgment of the practitioner and are particular to each individual. However, suitable dosage ranges for systemic application are disclosed herein and depend on the route of administration. Suitable regimes for administration are also variable but are typified by an initial administration followed by repeated doses at one or more hour intervals by a subsequent injection or other administration. Alternatively, continuous intravenous infusion sufficient to maintain concentrations in the blood in the ranges specified for in vivo therapies are contemplated.

In some embodiments, the methods further comprise administering a composition, CAL, or CAR, or cell described herein along with one or more additional autoimmune, GvHD, or transplant rejection agents, biologics, drugs, or treatments as part of a combinatorial therapy. Exemplary treatments for transplant rejection or GvHD include but are not limited to, Immunosuppressive drugs, e.g., Cyclosporine (Neoral, Sandimmune, Gengraf, and Restasis), Tacrolimus (Prograf, Protopic, Astagraf XL, and Envarsus XR), Methotrexate (Trexall, Rasuvo, Rheumatrex, and Otrexup (PF)), Sirolimus (Rapamune), Mycophenolic acid (Myfortic and CellCept), Rituximab (Rituxan), etanercept (Enbrel), pentostatin (Nipent), ruxolitinib (Jakafi); Chemotherapies, e.g., Methotrexate (Trexall, Rasuvo, Rheumatrex, and Otrexup (PF)), antithymocyte globulin (Atgam, Thymoglobulin); Steroids, e.g., Prednisone (Deltasone, Rayos, and Prednisone Intensol), Methylprednisolone (Medrol, Solu-Medrol, and Depo-Medrol), budesonide (Entocort EC, Uceris); Antifungal, e.g., Posaconazole (Noxafil); Antiviral drugs, e.g., Acyclovir (Zovirax and Sitavig), Valacyclovir (Valtrex); and Antibiotics, e.g., Sulfamethoxazole/Trimethoprim (Bactrim, Sulfatrim, and Bactrim DS); Protease inhibitors, e.g. alphal-proteinase inhibitor (Zemaira); extracorporeal photopheresis; monoclonal antibodies (daclizumab (Zinbryta), basiliximab (Simulect)), Brentuximab vedotin (Adcetris), Alemtuzumab (Campath, Lemtrada), Tocilizumab (Actemra); infusion of mesenchymal stromal cells.

Exemplary treatments for autoimmune disease include but are not limited to, Insulin, e.g., Insulin glulisine (Apidra and Apidra SoloStar), Insulin detemir (Levemir and Levemir FlexTouch), Insulin aspart (NovoLog, Novolog Flexpen, and Novolog PenFill), Insulin lispro (Humalog and Humalog KwikPen), Insulin, Insulin glargine (Lantus, Lantus Solostar, and Toujeo SoloStar); Dietary supplement, e.g., glucose tablets; and Hormones, e.g., Glucagon (GlucaGen and Glucagon Emergency Kit (human)), antidiabetic agents (Metformin (D-Care DM2, Fortamet, Glucophage, Glucophage XR, Glumetza, Riomet), glucagon-like peptide-1 (GLP-1) receptor agonist (liraglutide (Saxenda; Victoza) or semaglutide (Ozempic) or sodium-glucose co-transporter 2 (SGLT2) inhibitor: empagliflozin (Jardiance), canagliflozin (Invokana); sulfonylureas: glipizide (GlipiZIDE XL, Glucotrol, Glucotrol XL); Meglitinide Analogs: repaglinide (Prandin); Thiazolidinedione: pioglitazone (Actos); dipeptidyl peptidase-4 (DPP-4) inhibitors: Sitagliptin (Januvia), Saxagliptin (Onglyza), Linagliptin (Tradjenta), Alogliptin (Nesina)

The efficacy of a given treatment, e.g., for an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD, can be determined by the skilled clinician. However, a treatment is considered “effective treatment,” as the term is used herein, if any one or all of the signs or symptoms of are altered in a beneficial manner or other clinically accepted symptoms are improved, or even ameliorated, e.g., by at least 10% following treatment with an agent as described herein. Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or described herein.

An effective amount for the treatment of a disease means that amount which, when administered to a mammal in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease. Efficacy of an agent can be determined by assessing physical indicators of, for example autoimmune disease (e.g., result of an ANA), T cell mediated inflammation or immune response, malignant T cell condition, transplant rejection (e.g., high fever, tenderness at transplant site, etc.), or GvHD (e.g., redness, pain, or other symptoms at transplant site). Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dosage can vary depending upon the dosage form employed and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio LD50/ED50. Compositions and methods that exhibit large therapeutic indices are preferred. A therapeutically effective dose can be estimated initially from cell culture assays. Also, a dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the active ingredient(s), which achieves a half-maximal inhibition of symptoms) as determined in cell culture, or in an appropriate animal model. Levels in plasma can be measured, for example, by immunoassay, various DNA detection technologies, or high performance liquid chromatography. The effects of any particular dosage can be monitored by a suitable bioassay, e.g., assay to assess reaction following transplant, level of inflammation, ANA measurement, among others. The dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.

Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization, or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or are described herein. Treatment includes any treatment of a disease in an individual or an animal (some non-limiting examples include a human or an animal) and includes: (1) inhibiting the disease, e.g., preventing a worsening of symptoms (e.g. pain or inflammation); or (2) relieving the severity of the disease, e.g., causing regression of symptoms. An effective amount for the treatment of a disease means that amount which, when administered to a subject in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease. Efficacy of an agent can be determined by assessing physical indicators of a condition or desired response, (e.g. a reduction of inflammation, etc.). It is well within the ability of one skilled in the art to monitor efficacy of administration and/or treatment by measuring any one of such parameters, or any combination of parameters. Efficacy can be assessed in animal models of a condition described herein, for example treatment of autoimmune disease, transplant rejection or GVHD. When using an experimental animal model, efficacy of treatment is evidenced when a statistically significant change in a marker is observed, e.g. inflammation.

In some embodiments, the technology described herein relates to a pharmaceutical composition comprising a CAL and/or CAR, or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR, or a multi-component CAL and/or CAR) as described herein, and optionally a pharmaceutically acceptable carrier. In some embodiments, the active ingredients of the pharmaceutical composition comprise a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR or a multi-component CAL and/or CAR) as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist essentially of a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR or a multi-component CAL and/or CAR) as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist of a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as described herein.

Pharmaceutically acceptable carriers and diluents include saline, aqueous buffer solutions, solvents and/or dispersion media. The use of such carriers and diluents is well known in the art. Some non-limiting examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids (23) serum component, such as serum albumin, HDL and LDL; (22) C2-C12 alcohols, such as ethanol; and (23) other non-toxic compatible substances employed in pharmaceutical formulations. Wetting agents, coloring agents, release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservative and antioxidants can also be present in the formulation. The terms such as “excipient”, “carrier”, “pharmaceutically acceptable carrier” or the like are used interchangeably herein. In some embodiments, the carrier inhibits the degradation of the active agent as described herein.

In some embodiments, the pharmaceutical composition comprising a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as described herein can be a parenteral dose form. Since administration of parenteral dosage forms typically bypasses the patient's natural defenses against contaminants, parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions. In addition, controlled-release parenteral dosage forms can be prepared for administration of a patient, including, but not limited to, DUROS®-type dosage forms and dose-dumping.

Suitable vehicles that can be used to provide parenteral dosage forms of a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as disclosed within are well known to those skilled in the art. Examples include, without limitation: sterile water; water for injection USP; saline solution; glucose solution; aqueous vehicles such as but not limited to, sodium chloride injection, Ringer's injection, dextrose Injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and propylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate. Compounds that alter or modify the solubility of a pharmaceutically acceptable salt of an active ingredient can also be incorporated into the parenteral dosage forms of the disclosure, including conventional and controlled-release parenteral dosage forms.

Pharmaceutical compositions can also be formulated to be suitable for oral administration, for example as discrete dosage forms, such as, but not limited to, tablets (including without limitation scored or coated tablets), pills, caplets, capsules, chewable tablets, powder packets, cachets, troches, wafers, aerosol sprays, or liquids, such as but not limited to, syrups, elixirs, solutions or suspensions in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion, or a water-in-oil emulsion. Such compositions contain a predetermined amount of the pharmaceutically acceptable salt of the disclosed compounds, and may be prepared by methods of pharmacy well known to those skilled in the art. See generally, Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams, and Wilkins, Philadelphia PA. (2005).

Conventional dosage forms generally provide rapid or immediate drug release from the formulation. Depending on the pharmacology and pharmacokinetics of the drug, use of conventional dosage forms can lead to wide fluctuations in the concentrations of the drug in a patient's blood and other tissues. These fluctuations can impact a number of parameters, such as dose frequency, onset of action, duration of efficacy, maintenance of therapeutic blood levels, toxicity, side effects, and the like. Advantageously, controlled-release formulations can be used to control a drug's onset of action, duration of action, plasma levels within the therapeutic window, and peak blood levels. In particular, controlled- or extended-release dosage forms or formulations can be used to ensure that the maximum effectiveness of a drug is achieved while minimizing potential adverse effects and safety concerns, which can occur both from under-dosing a drug (i.e., going below the minimum therapeutic levels) as well as exceeding the toxicity level for the drug. In some embodiments, the composition can be administered in a sustained release formulation.

Controlled-release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled release counterparts. Ideally, the use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time. Advantages of controlled-release formulations include: 1) extended activity of the drug; 2) reduced dosage frequency; 3) increased patient compliance; 4) usage of less total drug; 5) reduction in local or systemic side effects; 6) minimization of drug accumulation; 7) reduction in blood level fluctuations; 8) improvement in efficacy of treatment; 9) reduction of potentiation or loss of drug activity; and 10) improvement in speed of control of diseases or conditions. Kim, Cherng-ju, Controlled Release Dosage Form Design, 2 (Technomic Publishing, Lancaster, Pa.: 2000).

Most controlled-release formulations are designed to initially release an amount of drug (active ingredient) that promptly produces the desired therapeutic effect, and gradually and continually release other amounts of drug to maintain this level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body. Controlled-release of an active ingredient can be stimulated by various conditions including, but not limited to, pH, ionic strength, osmotic pressure, temperature, enzymes, water, and other physiological conditions or compounds.

A variety of known controlled- or extended-release dosage forms, formulations, and devices can be adapted for use with the salts and compositions of the disclosure. Examples include, but are not limited to, those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,733,566; and 6,365,185 B1; each of which is incorporated herein by reference. These dosage forms can be used to provide slow or controlled-release of one or more active ingredients using, for example, hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems (such as OROS® (Alza Corporation, Mountain View, Calif. USA)), or a combination thereof to provide the desired release profile in varying proportions.

Described herein is a therapy called “CAL-cell therapy”, which seeks to help immune killer cells recognize autoreactive and alloreactive T cells. This is accomplished by genetically altering an immune cell so that it expresses a chimeric antigen ligand (CAL). The CAL is an altered ligand, in which the natural recognition portion is removed and replaced with a synthetic recognition portion, (including all synthetic or natural peptide MHC complexes) that is designed to more effectively recognize the autoreactive and alloreactive T cells by very specifically detecting the presence of a T cell receptor unique to the autoreactive and alloreactive T cells. These CAL immune cells are then given to a patient. Inside the patient, their synthetic CAL ligand molecules will bind to the autoreactive and alloreactive T cells and in the act of that binding, activate the killer immune cells, resulting in the engineered CAL-immune cells attacking the pathologic autoreactive and alloreactive T-cells to eliminate or reduce allo- or autoreactive immune response. This has particular applications for autoimmune conditions, cell-, tissue-, organ-transplants, and Graft vs. Host Disease (GvHD).

This invention is based, in part, on the finding that an engineered polypeptide presented herein can recognize and bind to the specific T cell receptor on the disease-causing T cells, deleting said T cells, e.g., eliminating or reducing autoimmune diseases or conditions; T cell mediated inflammation or immune response; and cell, tissue, organ transplants and Graft vs. Host Disease (GvHD),In some embodiments, the engineered polypeptide is composed of a peptide-major histocompatibility complex (pMHC) (e.g. as a monomer, oligomer, or multimer) as the recognition site for the TCR of an allogeneic or an autoreactive T cell. In some embodiments, the pMHC is one of the complexes described herein, e.g., see Tables 5 and 6.

In some embodiments, the engineered polypeptide is composed of a pMHC conjugated to a FITC, PE, or other biomolecular interaction domain described herein. The engineered polypeptide can also be considered as an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to a biomolecular interaction domain. The CAL technology described herein can target T cell clones in an antigen specific manner and is not dependent on any specific CAR construct to exhibit killing effect.

In some embodiments, the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as the recognition site fused to signaling domains from T cell receptors. Further provided herein is a split version of this CAR system, in which the CAR is composed of two pieces. The first piece is a universal CAR (also referred to herein as a Uni CAL) with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain, e.g., that is specific to each disease state. The second piece is an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to the cognate biomolecular interaction domain.

In some embodiments, the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as the recognition site fused to signaling domains from T cell receptors. Further provided herein is a split version of this CAR system, in which the CAR is composed of two pieces. The first piece is a universal CAR (also referred to herein as a Uni CAL) with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain, e.g., that is specific to each disease state. The second piece is an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to the cognate biomolecular interaction domain. It is noted that protein interaction domains are a type of biomolecular interaction domains and where one is specified herein, the other may always be substituted. The CAL technology described herein can target T cells clones in an antigen specific manner and is not dependent on any specific CAR construct to exhibit killing effect.

Accordingly, one aspect presented herein provides a composition comprising (a) chimeric antigen ligand or a TCR recognition domain; and one or both of (a) an intracellular signaling domain; and (b) a first-type protein interaction domain.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a chimeric antigen ligand or TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain.

In one embodiment of any aspect, the third-type protein interaction domain and first-type protein interaction domain have a higher affinity for each other than the second-type protein interaction domain and first-type protein interaction domain.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

In one embodiment of any aspect, the fourth-type protein interaction domain and fifth-type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first-type protein interaction domain.

In one embodiment of any aspect, the first polypeptide further comprises a sixth-type protein interaction domain and the recognition polypeptide further comprises a seventh-type protein interaction domain which bind specifically to each other.

In one embodiment of any aspect, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.

In one embodiment of any aspect, the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.

TCR recognition domains may not comprise a single polypeptide, but rather comprise two or more polypeptides and may additionally even comprise non-polypeptides. For example, a single MHC class II tetramer TCR recognition domain can comprise 4 biotin small molecules and 16 polypeptides (4 peptides, 4 MHC class II alpha chains, 4 MHC class II beta chains, and 4 streptavidin proteins. Other types of TCR recognition domains may additionally comprise other non-polypeptide molecules (e.g., MHC dextramers contain a polysaccharide backbone to which the MI-ICs are anchored to). In some embodiments, a composition described herein can comprise multiple copies or instances of a TCR recognition domain(s), e.g. the TCR recognition domain can be a mulitmer, or oligomer. In some embodiments, a composition described herein can comprise multiple copies or instances of a first polypeptide as described herein. In some embodiments, the first polypeptide comprises the entire TCR recognition domain. In some embodiments, the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.

In one embodiment of any aspect, the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; or a MHC-peptide fusion.

In one embodiment of any aspect, the peptide is a human, non-human, or synthetic/engineered peptide. The peptides can further comprise non-proteinaceous motifs, modifications, or domains, e.g., they can comprise glycosylation and/or lipids. In one embodiment of any aspect, the peptide is a Minor Histocompatibility Antigen (MiHA).

In one embodiment of any aspect, the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.

In one embodiment of any aspect, the protein interaction domains are found on an extracellular portion of the respective polypeptides.

In one embodiment of any aspect, (a) the protein interaction domain(s) is a leucine zipper, or any binding pair of protein interaction domains are collectively a pair of leucine zippers; (b) the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE); (c) the protein interaction domain(s) is a PSD95-Dlg1-zo-1 (PDZ) domain; (d) the protein interaction domain(s) is a streptavidin and/or a streptavidin binding protein (SBP) or any binding pair of protein interaction domains are collectively a streptavidin and a streptavidin binding protein (SBP); (e) the protein interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding protein (FKBP); (f) the protein interaction domain(s) is a cyclophilin-Fas fusion protein (CyP-Fas) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a cyclophilin-Fas fusion protein (CyP-Fas) and a FK506 binding protein (FKBP); (g) the protein interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a calcineurin A (CNA) and a FK506 binding protein (FKBP); (h) the protein interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of protein interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1); (i) the protein interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of protein interaction domains are collectively a Snap-tag and a Halo tag; (j) the protein interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of protein interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52); (k) the protein interaction domain(s) is a PYL and/or a ABI, or any binding pair of protein interaction domains are collectively a PYL and a ABI; (1) the protein interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of protein interaction domains are collectively a nucleotide tag and a zinc finger domain; (m) wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags; (n) the protein interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding protein or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or (o) the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein.

In one embodiment of any aspect, the nucleotide tag is a DNA tag or dsDNA tag.

In one embodiment of any aspect, the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of: TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.

In one embodiments of any aspect, a cell comprising and/or expressing a comprising a composition comprising a TCR recognition domain and an intracellular signaling domain further comprises a TCR signaling-responsive promoter operatively linked to a payload transgene. Such embodiments permit transgene payload expression specifically to and/or in the vicinity of a targeted T cell. Suitable promoters and transgene are known in the art, e.g., those promoters and transgenes used in “TRUCK CAR” technology. An exemplary promoter is a NFAT-sensitive promoter. Exemplary transgene payloads can include checkpoint inhibitors (e.g., CTLA-4, [Ipilimumab, Tremelimumab] or PD-1 [Nivolumab, Pembrolizumab, Pidilizumab]) or proinflammatory cytokines (e.g., IL-2, IL-12, etc). In our case this will be used for in-situ targeting of a patient's anticancer T-cells to very specifically and locally deliver activating agents like one or more checkpoint inhibitors (CTLA-4, [Ipilimumab, Tremelimumab] or PD-1 [Nivolumab, Pembrolizumab, Pidilizumab]) and/or proinflammatory cytokines, (e.g. IL-2, IL-12, etc.) that can push them to expansion and effector phenotype. With this targeted delivery, it is expected to see more efficacy without the systemic side effects caused by checkpoint inhibitors and cytokines administered systemically. Further, combination therapy with multiple different agents is possible with the described technolog as there is minimal systemic side effect due to paracrine delivery. Further description of suitable promoters, payloads, as well as how to make and use TRUCK technology is described, e.g., Peterson et al. Front. Oncol. 2019 9:69; Chmielewski et al. Advances in Cell and Gene Therapy 2020 3:e84; Chimielewski et al. Expert opinion Biol Ther 2015 15:1145-54; and Chimieleski et al. Immunol Rev 2014 257:83-90; each of which is incorporated by reference herein in its entirety. In some embodiments, the cell can be allogeneic, e.g., and engineered once and given as pulse therapy. In some embodiments, the cell can be a T cell or any other cell type described herein, e.g., a NK cell. Exemplary, non-limiting proinflammatory cytokines include IFNs, IFN-γ, TNFα, TGF-β, IL-1β, IL-6, IL-4, IL-10, IL-13, IL-2, IL-12, IL-15, and IL-27.

A promoter can be said to drive expression or drive transcription of the nucleic acid sequence that it regulates. The phrases “operably linked,” “operatively positioned,” “operatively linked,” “under control,” and “under transcriptional control” indicate that a promoter is in a correct functional location and/or orientation in relation to a nucleic acid sequence it regulates to control transcriptional initiation and/or expression of that sequence.

In some embodiments, a composition comprising a TCR recognition domain and an biomolecular interaction domain is a soluble molecule and/or soluble complex.

Another aspect provided herein is a cell comprising and/or expressing the composition of any of the compositions described herein.

In one embodiment of any aspect, the TCR recognition domain comprises a MHC allogeneic to the cell comprising and/or expressing the composition. In one embodiment of any aspect, the TCR recognition domain comprises a MHC allogeneic to the cell that the TCR originated from.

In one embodiment of any aspect, the TCR recognition domain comprises a peptide allogeneic to the cell comprising and/or expressing the composition. In one embodiment of any aspect, the TCR recognition domain comprises a peptide allogeneic to the cell that the TCR originated from.

In one embodiment of any aspect, the cell is a dendritic cell (CAL DC), a T cell (e.g., effector, regulatory, etc.,) (CAL-T); regulatory T cell, effector T cell, natural killer cell (CAL NK), or any other myeloid cell. In one embodiment of any aspect, the cell is engineered to express the polypeptide(s) of the composition. In one embodiment of any aspect, the cell is engineered to express the signaling polypeptide of the composition. In one embodiment of any aspect, the cell is further engineered to knockout the native MHCI/II. In one embodiment of any aspect, the cell is further engineered to lack cell surface expression of native MHCI/II.

Another aspect provided herein is a chimeric antigen receptor (CAR) comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) an intracellular signaling domain.

Another aspect provided herein is a composition comprising a first polypeptide comprising: (a) an anti-CD127 and/or anti-CD45RO recognition domain; (b) a first-type protein interaction domain; and a second polypeptide comprising (a) a second-type protein interaction domain; and (b) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.

Another aspect provided herein is a composition comprising a first polypeptide comprising (a) an anti-CD127 recognition domain; (b) a first-type protein interaction domain; a second polypeptide comprising (a) an anti-CD45RO recognition domain; (b) a fifth-type protein interaction domain; and a third polypeptide comprising (a) a second-type and a fourth-type protein interaction domain; and (b) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

Another aspect provided herein is a cell comprising any of the CARs described herein, or any of the compositions described herein.

Another aspect provided herein is a method of preventing and/or treating an autoimmune diseases or conditions or T cell mediated inflammation or immune response; or treating or preventing transplant rejection or GvHD in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein. Another aspect provided herein is a method of preventing and/or treating a malignant T cell condition in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein.

In various embodiments of any aspect, the TCR recognition domain comprises a MHC allogeneic to the subject, a MHC autologous to the transplant cells, a peptide allogeneic to the subject, or a peptide autologous to the transplant cells.

In one embodiment of any aspect, the transplant is vascularized composite allotransplantation (VCA).

In one embodiment of any aspect, the autoimmune disease is type 1 diabetes, multiple sclerosis, rheumatoid arthritis, or scleroderma.

One aspect of the embodiments provided herein is a CAR T cell that targets a CD127+/CD45RO+ T cell. In one embodiment, the CD127+/CD45RO+ T cell is a CD127+/CD45RO+ memory T cell. In one embodiment, the CD127+/CD45RO+ T cell is an alloreactive CD127+/CD45RO+ T cell.

Accordingly, one aspect herein provides a CAR comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) intracellular signaling domain. Further provided herein is a composition comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) intracellular signaling domain.

Another aspect of the provides a composition comprising a first polypeptide comprising: (a) an anti-CD127 and/or anti-CD45RO recognition domain; (b) a first-type protein interaction domain; and a second polypeptide comprising: (c) a second-type protein interaction domain; and (d) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.

Another aspect of the provides a composition comprising a first polypeptide comprising: (a) an anti-CD127 recognition domain; (b) a first-type protein interaction domain; a second polypeptide comprising: (c) an anti-CD45RO recognition domain; (d) a fifth-type protein interaction domain; and a third polypeptide comprising: (e) a second-type and a fourth-type protein interaction domain; and (f) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

In one embodiment, the anti-CD127 recognition domain recognizes and binds to the sequence of CD127 on a CD127+ cell, e.g., a CD127+/CD45RO+ T cell. As used herein, “CD127”, also referred to as Interleukin 7 receptor a (IL7Ra), ILRA, IL7RA, CDW127, or IL-7R-alpha, is a cell surface receptor that has been shown to have a role in V(D)J recombination during lymphocyte development. Defects in CD127 have been associated with severe combined immunodeficiency (SCID). Sequences for CD127 are known for a number of species, e.g., human CD127 (NCBI Gene ID: 3575), mRNA (NCBI Ref Seq: NM_002185.5), and polypeptide (NCBI Ref Seq: NP_002176.2). CD127 refers to all naturally occurring variants or isoforms of CD127. In one embodiment, the CD127 polypeptide sequence is presented in SEQ ID NO: 1. In some embodiments of any of the aspects, the CD127 polypeptide can be an ortholog, variant, and/or allele of SEQ ID NO: 1.

(SEQ ID NO: 1)

1	MTILGTTFGM VFSLLQVVSG ESGYAQNGDL EDAELDDYSF SCYSQLEVNG SQHSLTCAFE
61	DPDVNITNLE FEICGALVEV KCLNFRKLQE IYFIETKKFL LIGKSNICVK VGEKSLTCKK
121	IDLTTIVKPE APFDLSVVYR EGANDEVVTF NTSHLQKKYV KVLMHDVAYR QEKDENKWTH
181	VNLSSTKLTL LQRKLQPAAM YEIKVRSIPD HYFKGFWSEW SPSYYFRTPE INNSSGEMDP
241	ILLTISILSF FSVALLVILA CVLWKKRIKP IVWPSLPDHK KTLEHLCKKP RKNLNVSFNP
301	ESFLDCQIHR VDDIQARDEV EGFLQDTFPQ QLEESEKQRL GGDVQSPNCP SEDVVITPES
361	FGRDSSLTCL AGNVSACDAP ILSSSRSLDC RESGKNGPHV YQDLLLSLGT TNSTLPPPFS
421	LQSGILTLNP VAQGQPILTS LGSNQEEAYV TMSSFYQNQ

In one embodiment, the anti-CD45RO recognition domain recognizes and binds to the sequence of CD45RO on a CD45RO+ cell, e.g., a CD127+/CD45RO+ T cell. As used herein, “CD45RO”, also referred to as PTPRC, LCA, LYS, B220, CD45, L-CA, T200, CD45R, and GP180, is to a cell surface signaling molecule that been shown to be an essential regulator of T- and B-cell antigen receptor signaling. Sequences for CD45RO are known for a number of species, e.g., human CD45RO (NCBI Gene ID: 5788), mRNA (NCBI Ref Seq: NM_001267798.2), and polypeptide (NCBI Ref Seq: NP_001254727.1). CD45RO refers to all naturally occurring variants or isoforms of CD45RO. In one embodiment, the CD45RO polypeptide sequence is presented in SEQ ID NO: 2. In some embodiments of any of the aspects, the CD45RO polypeptide can be an ortholog, variant, and/or allele of SEO ID NO: 2.

(SEQ ID NO: 2)

1	MTMYLWLKLL AFGFAFLDTE VFVTGQSPTP SPTGHLQAEE QGSQSKSPNL KSREADSSAF
61	SWWPKAREPL TNHWSKSKSP KAEELGV

For convenience, the meaning of some terms and phrases used in the specification, examples, and appended claims, are provided below. Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is an apparent discrepancy between the usage of a term in the art and its definition provided herein, the definition provided within the specification shall prevail.

For convenience, certain terms employed herein, in the specification, examples and appended claims are collected here.

The terms “decrease”, “reduced”, “reduction”, or “inhibit” are all used herein to mean a decrease by a statistically significant amount. In some embodiments, “reduce,” “reduction” or “decrease” or “inhibit” typically means a decrease by at least 10% as compared to a reference level (e.g. the absence of a given treatment) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, “reduction” or “inhibition” does not encompass a complete inhibition or reduction as compared to a reference level. “Complete inhibition” is a 100% inhibition as compared to a reference level. A decrease can be preferably down to a level accepted as within the range of normal for an individual without a given disorder.

The terms “increased”, “increase”, “enhance”, or “activate” are all used herein to mean an increase by a statically significant amount. In some embodiments, the terms “increased”, “increase”, “enhance”, or “activate” can mean an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level. In the context of a marker or symptom, a “increase” is a statistically significant increase in such level.

As used herein, a “subject” means a human or animal. Usually the animal is a vertebrate such as a swine, primate, rodent, domestic animal or game animal. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents include mice, rats, woodchucks, ferrets, rabbits and hamsters. Domestic and game animals include cows, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon. In some embodiments, the subject is a mammal, e.g., a primate, e.g., a human. The terms, “individual,” “patient” and “subject” are used interchangeably herein.

Preferably, the subject is a mammal. The mammal can be a human, non-human swine, primate, mouse, rat, dog, cat, horse, or cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects that represent animal models of autoimmune diseases or conditions; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. A subject can be male or female.

A subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment (e.g. an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD) or one or more complications related to such a condition, and optionally, have already undergone treatment for an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or the one or more complications related to an autoimmune disease, transplant rejection, or GvHD. Alternatively, a subject can also be one who has not been previously diagnosed as having an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or one or more complications related to an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. For example, a subject can be one who exhibits one or more risk factors for an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or one or more complications related to an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or a subject who does not exhibit risk factors.

A “subject in need” of treatment for a particular condition can be a subject having that condition, diagnosed as having that condition, or at risk of developing that condition, e.g., an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD.

In some embodiments, a nucleic acid encoding a CAL, CAR, a multi-component CAL and/or CAR or portion thereof as described herein is comprised by a vector. In some of the aspects described herein, a nucleic acid sequence encoding a multi-component CAL and/or CAR, or portion thereof as described herein, or any module thereof, is operably linked to a vector. The term “vector”, as used herein, refers to a nucleic acid construct designed for delivery to a host cell or for transfer between different host cells. As used herein, a vector can be viral or non-viral. The term “vector” encompasses any genetic element that is capable of replication when associated with the proper control elements and that can transfer gene sequences to cells. A vector can include, but is not limited to, a cloning vector, an expression vector, a plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc.

As used herein, the term “expression vector” refers to a vector that directs expression of an RNA or polypeptide from sequences linked to transcriptional regulatory sequences on the vector. The sequences expressed will often, but not necessarily, be heterologous to the cell. An expression vector may comprise additional elements, for example, the expression vector may have two replication systems, thus allowing it to be maintained in two organisms, for example in human cells for expression and in a prokaryotic host for cloning and amplification. The term “expression” refers to the cellular processes involved in producing RNA and proteins and as appropriate, secreting proteins, including where applicable, but not limited to, for example, transcription, transcript processing, translation and protein folding, modification and processing. “Expression products” include RNA transcribed from a gene, and polypeptides obtained by translation of mRNA transcribed from a gene. The term “gene” means the nucleic acid sequence that is transcribed (DNA) to RNA in vitro or in vivo when operably linked to appropriate regulatory sequences. The gene may or may not include regions preceding and following the coding region, e.g. 5′ untranslated (5′UTR) or “leader” sequences and 3′ UTR or “trailer” sequences, as well as intervening sequences (introns) between individual coding segments (exons).

As used herein, the term “viral vector” refers to a nucleic acid vector construct that includes at least one element of viral origin and has the capacity to be packaged into a viral vector particle. The viral vector can contain the nucleic acid encoding a CAL and/or CAR described herein, e.g., a multi-component CAL and/or CAR, or portion thereof as described herein in place of non-essential viral genes. The vector and/or particle may be utilized for the purpose of transferring any nucleic acids into cells either in vitro or in vivo. Numerous forms of viral vectors are known in the art.

By “recombinant vector” is meant a vector that includes a heterologous nucleic acid sequence, or “transgene” that is capable of expression in vivo or in the transduced cells. It should be understood that the vectors described herein can, in some embodiments, be combined with other suitable compositions and therapies. In some embodiments, the vector is episomal. The use of a suitable episomal vector provides a means of maintaining the nucleotide of interest in the subject in high copy number extra chromosomal DNA thereby eliminating potential effects of chromosomal integration.

As used herein, the term “nucleic acid” or “nucleic acid sequence” refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid or an analog thereof. The nucleic acid can be either single-stranded or double-stranded. A single-stranded nucleic acid can be one nucleic acid strand of a denatured double-stranded DNA. Alternatively, it can be a single-stranded nucleic acid not derived from any double-stranded DNA. In one aspect, the nucleic acid can be DNA. In another aspect, the nucleic acid can be RNA, e.g., single-stranded or double-stranded RNA. Suitable nucleic acid molecules are DNA, including genomic DNA or cDNA. Other suitable nucleic acid molecules are RNA, including mRNA.

As used herein, the terms “protein” and “polypeptide” are used interchangeably herein to designate a series of amino acid residues, connected to each other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues. The terms “protein”, and “polypeptide” refer to a polymer of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogs, regardless of its size or function. “Protein” and “polypeptide” are often used in reference to relatively large polypeptides, whereas the term “peptide” is often used in reference to small polypeptides, but usage of these terms in the art overlaps. The terms “protein” and “polypeptide” are used interchangeably herein when referring to a gene product and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments and other equivalents, variants, fragments, and analogs of the foregoing.

As used herein an “antibody” refers to IgG, IgM, IgA, IgD or IgE molecules or antigen-specific antibody fragments thereof (including, but not limited to, a Fab, F(ab′)2, Fv, disulphide linked Fv, scFv, single domain antibody, closed conformation multispecific antibody, disulphide-linked scfv, diabody), whether derived from any species that naturally produces an antibody, or created by recombinant DNA technology; whether isolated from serum, B-cells, hybridomas, transfectomas, yeast or bacteria.

As described herein, an “antigen” is a molecule that is bound by a binding site on an antibody agent. Typically, antigens are bound by antibody ligands and are capable of raising an antibody response in vivo. An antigen can be a polypeptide, protein, nucleic acid or other molecule or portion thereof. The term “antigenic determinant” refers to an epitope on the antigen recognized by an antigen-binding molecule, and more particularly, by the antigen-binding site of said molecule.

As used herein, the term “antibody reagent” refers to a polypeptide that includes at least one immunoglobulin variable domain or immunoglobulin variable domain sequence and which specifically binds a given antigen. An antibody reagent can comprise an antibody or a polypeptide comprising an antigen-binding domain of an antibody. In some embodiments, an antibody reagent can comprise a monoclonal antibody or a polypeptide comprising an antigen-binding domain of a monoclonal antibody. For example, an antibody can include a heavy (H) chain variable region (abbreviated herein as VH), and a light (L) chain variable region (abbreviated herein as VL). In another example, an antibody includes two heavy (H) chain variable regions and two light (L) chain variable regions. The term “antibody reagent” encompasses antigen-binding fragments of antibodies (e.g., single chain antibodies, Fab and sFab fragments, F(ab′)2, Fd fragments, Fv fragments, scFv, and domain antibodies (dAb) fragments (see, e.g. de Wildt et al., Eur J. Immunol. 1996; 26(3):629-39; which is incorporated by reference herein in its entirety)) as well as complete antibodies. An antibody can have the structural features of IgA, IgG, IgE, IgD, IgM (as well as subtypes and combinations thereof). Antibodies can be from any source, including mouse, rabbit, pig, rat, and primate (human and non-human primate) and primatized antibodies. Antibodies also include midibodies, humanized antibodies, chimeric antibodies, and the like.

The VH and VL regions can be further subdivided into regions of hypervariability, termed “complementarity determining regions” (“CDR”), interspersed with regions that are more conserved, termed “framework regions” (“FR”). The extent of the framework region and CDRs has been precisely defined (see, Kabat, E. A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242, and Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917; which are incorporated by reference herein in their entireties). Each VH and VL is typically composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

The terms “antigen-binding fragment” or “antigen-binding domain”, which are used interchangeably herein are used to refer to one or more fragments of a full length antibody that retain the ability to specifically bind to a target of interest. Examples of binding fragments encompassed within the term “antigen-binding fragment” of a full length antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment including two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546; which is incorporated by reference herein in its entirety), which consists of a VH or VL domain; and (vi) an isolated complementarity determining region (CDR) that retains specific antigen-binding functionality.

As used herein, the term “specific binding” refers to a chemical interaction between two molecules, compounds, cells and/or particles wherein the first entity binds to the second, target entity with greater specificity and affinity than it binds to a third entity which is a non-target. In some embodiments, specific binding can refer to an affinity of the first entity for the second target entity which is at least 10 times, at least 50 times, at least 100 times, at least 500 times, at least 1000 times or greater than the affinity for the third nontarget entity. A reagent specific for a given target is one that exhibits specific binding for that target under the conditions of the assay being utilized. In some embodiments, binding described herein can be preferential binding, e.g., binding between two molecules, compounds, cells and/or particles wherein the first entity binds to the second, target entity with at least 2 times greater specificity and affinity than it binds to a third entity which is a non-target.

Additionally, and as described herein, a recombinant humanized antibody can be further optimized to decrease potential immunogenicity, while maintaining functional activity, for therapy in humans. In this regard, functional activity means a polypeptide capable of displaying one or more known functional activities associated with a recombinant antibody or antibody reagent thereof as described herein. Such functional activities include, e.g. the ability to bind to a target.

As used herein, the terms “treat,” “treatment,” “treating,” or “amelioration” refer to therapeutic treatments, wherein the object is to reverse, alleviate, ameliorate, inhibit, slow down or stop the progression or severity of a condition associated with a disease or disorder, e.g. autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. The term “treating” includes reducing or alleviating at least one adverse effect or symptom of a condition, disease or disorder associated with a, e.g. autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of a disease is reduced or halted. That is, “treatment” includes not just the improvement of symptoms or markers, but also a cessation of, or at least slowing of, progress or worsening of symptoms compared to what would be expected in the absence of treatment. Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, remission (whether partial or total), and/or decreased mortality, whether detectable or undetectable. The term “treatment” of a disease also includes providing relief from the symptoms or side-effects of the disease (including palliative treatment).

As used herein, the term “pharmaceutical composition” refers to the active agent in combination with a pharmaceutically acceptable carrier e.g. a carrier commonly used in the pharmaceutical industry. The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

As used herein, the term “administering,” refers to the placement of an agent, e.g., a CAL, CAR, composition, or cell as disclosed herein into a subject by a method or route which results in at least partial delivery of the agent at a desired site. Pharmaceutical compositions comprising the compounds disclosed herein can be administered by any appropriate route which results in an effective treatment in the subject.

Immune checkpoint inhibitors inhibit one or more immune checkpoint proteins. The immune system has multiple inhibitory pathways that are critical for maintaining self-tolerance and modulating immune responses. For example, in T-cells, the amplitude and quality of response is initiated through antigen recognition by the T-cell receptor and is regulated by immune checkpoint proteins that balance co-stimulatory and inhibitory signals. In some embodiments of any of the aspects, a subject or patient is treated with at least one inhibitor of an immune checkpoint protein. As used herein, “immune checkpoint protein” refers to a protein which, when active, exhibits an inhibitory effect on immune activity, e.g., T cell activity. Exemplary immune checkpoint proteins can include PD-1 (e.g., NCBI Gene ID: 5133); PD-L1 (e.g., NCBI Gene ID: 29126); PD-L2 (e.g., NCBI Gene ID: 80380); TIM-3 (e.g., NCBI Gene ID: 84868); CTLA4 (e.g., NCBI Gene ID: 1493); TIGIT (e.g., NCBI Gene ID: 201633); KIR (e.g., NCBI Gene ID: 3811); LAG3 (e.g., NCBI Gene ID: 3902); DD1-α (e.g., NCBI Gene ID: 64115); A2AR (e.g., NCBI Gene ID: 135); B7-H3 (e.g., NCBI Gene ID: 80381); B7-H4 (e.g., NCBI Gene ID: 79679); BTLA (e.g., NCBI Gene ID: 151888); IDO (e.g., NCBI Gene ID: 3620); TDO (e.g., NCBI Gene ID: 6999); HVEM (e.g., NCBI Gene ID: 8764); GALS (e.g., NCBI Gene ID: 3965); 2B4 (belongs to the CD2 family of molecules and is expressed on all NK, γδ, and memory CD8+(αβ) T cells) (e.g., NCBI Gene ID: 51744); CD160 (also referred to as BY55) (e.g., NCBI Gene ID: 11126); and various B-7 family ligands. B7 family ligands include, but are not limited to, B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6 and B7-H7.

Non-limiting examples of immune checkpoint inhibitors (with checkpoint targets and manufacturers noted in parantheses) can include: MGA271 (B7-H3: MacroGenics); ipilimumab (CTLA-4; Bristol Meyers Squibb); pembrolizumab (PD-1; Merck); nivolumab (PD-1; Bristol Meyers Squibb); atezolizumab (PD-L1; Genentech); galiximab (B7.1; Biogen); IMP321 (LAG3: Immuntep); BMS-986016 (LAG3; Bristol Meyers Squibb); SMB-663513 (CD137; Bristol-Meyers Squibb); PF-05082566 (CD137; Pfizer); IPH2101 (KIR; Innate Pharma); KW-0761 (CCR4; Kyowa Kirin); CDX-1127 (CD27; CellDex); MEDI-6769 (Ox40; MedImmune); CP-870,893 (CD40; Genentech); tremelimumab (CTLA-4; Medimmune); pidilizumab (PD-1; Medivation); MPDL3280A (PD-L1; Roche); MEDI4736 (PD-L1; AstraZeneca); MSB0010718C (PD-L1; EMD Serono); AUNP12 (PD-1; Aurigene); avelumab (PD-L1; Merck); durvalumab (PD-L1; Medimmune); IMP321, a soluble Ig fusion protein (Brignone et al., 2007, J. Immunol. 179:4202-4211); the anti-B7-H3 antibody MGA271 (Loo et al., 2012, Clin. Cancer Res. July 15 (18) 3834); TIM3 (T-cell immunoglobulin domain and mucin domain 3) inhibitors (Fourcade et al., 2010, J. Exp. Med. 207:2175-86 and Sakuishi et al., 2010, J. Exp. Med. 207:2187-94); anti-CTLA-4 antibodies described in U.S. Pat. Nos. 5,811,097; 5,811,097; 5,855,887; 6,051,227; 6,207,157; 6,682,736; 6,984,720; and 7,605,238; tremelimumab, (ticilimumab, CP-675,206); ipilimumab (also known as 10D1, MDX-D010); PD-1 and PD-L1 blockers described in U.S. Pat. Nos. 7,488,802; 7,943,743; 8,008,449; 8,168,757; 8,217,149, and PCT Published Patent Application Nos: WO03042402, WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400, and WO2011161699; nivolumab (MDX 1106, BMS 936558, ONO 4538); lambrolizumab (MK-3475 or SCH 900475); CT-011; AMP-224; and BMS-936559 (MDX-1105-01). The foregoing references are incorporated by reference herein in their entireties.

The term “statistically significant” or “significantly” refers to statistical significance, e.g., a rejection of the null hypothesis with a p-value of less than 0.05 and generally means a two standard deviation (2SD) or greater difference.

Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term “about.” The term “about” when used in connection with percentages can mean±1%.

As used herein the term “comprising” or “comprises” is used in reference to compositions, methods, and respective component(s) thereof, that are essential to the method or composition, yet open to the inclusion of unspecified elements, whether essential or not.

The term “consisting of” refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.

As used herein the term “consisting essentially of” refers to those elements required for a given embodiment. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment.

The singular terms “a,” “an,” and “the” include plural referents unless context clearly indicates otherwise. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. The abbreviation, “e.g.” is derived from the Latin exempli gratia and is used herein to indicate a non-limiting example. Thus, the abbreviation “e.g.” is synonymous with the term “for example.”

Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined solely by the claims. Definitions of common terms in immunology and molecular biology can be found in The Merck Manual of Diagnosis and Therapy, 19th Edition, published by Merck Sharp & Dohme Corp., 2011 (ISBN 978-0-911910-19-3); Robert S. Porter et al. (eds.), The Encyclopedia of Molecular Cell Biology and Molecular Medicine, published by Blackwell Science Ltd., 1999-2012 (ISBN 9783527600908); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8); Immunology by Werner Luttmann, published by Elsevier, 2006; Janeway's Immunobiology, Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), Taylor & Francis Limited, 2014 (ISBN 0815345305, 9780815345305); Lewin's Genes XI, published by Jones & Bartlett Publishers, 2014 (ISBN-1449659055); Michael Richard Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (2012) (ISBN 1936113414); Davis et al., Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (2012) (ISBN 044460149X); Laboratory Methods in Enzymology: DNA, Jon Lorsch (ed.) Elsevier, 2013 (ISBN 0124199542); Current Protocols in Molecular Biology (CPMB), Frederick M. Ausubel (ed.), John Wiley and Sons, 2014 (ISBN 047150338X, 9780471503385), Current Protocols in Protein Science (CPPS), John E. Coligan (ed.), John Wiley and Sons, Inc., 2005; and Current Protocols in Immunology (CPI) (John E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach, Warren Strobe, (eds.) John Wiley and Sons, Inc., 2003 (ISBN 0471142735, 9780471142737), the contents of which are all incorporated by reference herein in their entireties.

Other terms are defined herein within the description of the various aspects of the invention.

All patents and other publications; including literature references, issued patents, published patent applications, and co-pending patent applications; cited throughout this application are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that might be used in connection with the technology described herein. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

Specific elements of any of the foregoing embodiments can be combined or substituted for elements in other embodiments. Furthermore, while advantages associated with certain embodiments of the disclosure have been described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all embodiments need necessarily exhibit such advantages to fall within the scope of the disclosure.

All patents and other publications identified in the specification and examples are expressly incorporated herein by reference for all purposes. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the invention and these are therefore considered to be within the scope of the invention as defined in the claims which follow. Further, to the extent not already indicated, it will be understood by those of ordinary skill in the art that any one of the various embodiments herein described and illustrated can be further modified to incorporate features shown in any of the other embodiments disclosed herein.

Some embodiments of the technology described herein can be defined according to any of the following numbered paragraphs:

• • 1. A composition, comprising:
    • a. a TCR recognition domain; and
    • one or both of:
    • b. an intracellular signaling domain; and
    • c. a first-type protein interaction domain.
  • 2. A composition comprising:
    • a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and
    • b. a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain;
    • wherein the first-type and second-type protein interaction domains bind specifically to each other.
  • 3. A composition comprising:
    • a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and
    • b. a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain;
    • wherein the first-type and third-type protein interaction domains bind specifically to each other.
  • 4. A composition comprising:
    • a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and
    • b. a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and
    • c. a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain;
    • wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain.
  • 5. The composition of any of paragraphs 3-4, wherein the third-type protein interaction domain and first-type protein interaction domain have a higher affinity for each other than the second-type protein interaction domain and first-type protein interaction domain.
  • 6. A composition comprising:
    • a. a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain;
    • b. a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and
    • c. a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain;
    • wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and
    • wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
  • 7. The composition of paragraph 6, wherein the fourth-type protein interaction domain and fifth-type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first-type protein interaction domain.
  • 8. The composition of any of paragraphs 6-7, wherein the first polypeptide further comprises a sixth-type protein interaction domain and the recognition polypeptide further comprises a seventh-type protein interaction domain which bind specifically to each other.
  • 9. The composition of any of paragraphs 3-8, wherein the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.
  • 10. The composition of any of paragraphs 3-9, wherein the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
  • 11. The composition of any of the preceding paragraphs, wherein the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; or a MHC-peptide fusion.
  • 12. The composition of paragraph 11, wherein the peptide is a human or non-human peptide.
  • 13. The composition of any of paragraphs 11-12, wherein the peptide is a Minor Histocompatibility Antigen (MiHA).
  • 14. The composition of any of paragraphs 11-13, wherein the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 15. The composition of any of the preceding paragraphs, wherein the protein interaction domains are found on an extracellular portion of the respective polypeptides.
  • 16. The composition of any of the preceding paragraphs,
    • a. wherein the protein interaction domain(s) is a leucine zipper, or any binding pair of protein interaction domains are collectively a pair of leucine zippers;
    • b. wherein the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE);
    • c. wherein the protein interaction domain(s) is a PSD95-Dlg1-zo-1 (PDZ) domain;
    • d. wherein the protein interaction domain(s) is a streptavidin and/or a streptavidin binding protein (SBP) or any binding pair of protein interaction domains are collectively a streptavidin and a streptavidin binding protein (SBP);
    • e. wherein the protein interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding protein (FKBP);
    • f. wherein the protein interaction domain(s) is a cyclophilin-Fas fusion protein (CyP-Fas) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a cyclophilin-Fas fusion protein (CyP-Fas) and a FK506 binding protein (FKBP);
    • g. wherein the protein interaction domain(s) is a calcineurinA (CNA) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a calcineurinA (CNA) and a FK506 binding protein (FKBP);
    • h. wherein the protein interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of protein interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1);
    • i. wherein the protein interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of protein interaction domains are collectively a Snap-tag and a Halo tag;
    • j. wherein the protein interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of protein interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52);
    • k. wherein the protein interaction domain(s) is a PYL and/or a ABI, or any binding pair of protein interaction domains are collectively a PYL and a ABI; and/or
    • l. wherein the protein interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of protein interaction domains are collectively a nucleotide tag and a zinc finger domain.
  • 17. The composition of paragraph 16, wherein the nucleotide tag is a DNA tag or dsDNA tag.
  • 18. The composition of any of the preceding paragraphs, wherein the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of:
    • TCRC; FcRγ; FcRp; CD3γ; CD35; CD3s; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; ZAP70; and 41BB.
  • 19. A cell comprising and/or expressing the composition of any of the preceding paragraphs.
  • 20. The cell of paragraph 19, wherein the TCR recognition domain comprises a MHC allogenic to the cell.
  • 21. The cell of paragraph 19, wherein the TCR recognition domain comprises a peptide allogenic to the cell.
  • 22. The cell of any of paragraphs 19-21, wherein the cell is a dendritic cell, regulatory T cell, or effector T cell.
  • 23. The cell of any of paragraphs 19-22, wherein the cell is engineered to express the polypeptide(s) of the composition.
  • 24. The cell of any of paragraphs 19-22, wherein the cell is engineered to express the signaling polypeptide of the composition.
  • 25. The cell of any of paragraphs 19-24, wherein the cell is further engineered to knockout the native MHCI/II.
  • 26. A chimeric antigen receptor (CAR) comprising:
    • a. an anti-CD127 and/or anti-CD45RO recognition domain;
    • b. an intracellular signaling domain.
  • 27. A composition comprising:
    • a first polypeptide comprising:
    • a. an anti-CD127 and/or anti-CD45RO recognition domain;
    • b. a first-type protein interaction domain; and
    • a second polypeptide comprising:
    • c. a second-type protein interaction domain; and
    • d. an intracellular signaling domain;
    • wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.
  • 28. A composition comprising:
    • a first polypeptide comprising:
    • a. an anti-CD127 recognition domain;
    • b. a first-type protein interaction domain;
    • a second polypeptide comprising:
    • c. an anti-CD45RO recognition domain;
    • d. a fifth-type protein interaction domain; and
    • a third polypeptide comprising:
    • e. a second-type and a fourth-type protein interaction domain; and
    • f. an intracellular signaling domain;
    • wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and
    • wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
  • 29. A cell comprising the CAR or composition of any of paragraphs 26-28.
  • 30. A method of treating an autoimmune disease or treating or preventing transplant rejection or GVHD in a subject in need thereof, the method comprising administering to the subject a composition and/or cell of any of the preceding paragraphs.
  • 31. The method of paragraph 30, wherein the TCR recognition domain comprises a MHC allogenic to the subject.
  • 32. The method of paragraph 30, wherein the TCR recognition domain comprises a MHC autologous to the transplant cells.
  • 33. The method of paragraph 30, wherein the TCR recognition domain comprises a peptide allogenic to the subject.
  • 34. The method of paragraph 30, wherein the TCR recognition domain comprises a peptide autologous to the transplant cells.
  • 35. The method of any of paragraphs 32 or 34, wherein the transplant is vascularized composite allotransplantation (VCA).
  • 36. The method of any of paragraphs 35, wherein the autoimmune disease is type 1 diabetes, multiple sclerosis, rheumatoid arthritis, or scleroderma.

Some embodiments of the technology described herein can be defined according to any of the following numbered paragraphs:

• • 1. A composition, comprising:
    • a. a TCR recognition domain; and
    • one or both of:
    • b. an intracellular signaling domain; and
    • c. a first-type biomolecular interaction domain.
  • 2. A composition comprising:
    • a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and
    • b. a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain;
    • wherein the first-type and second-type biomolecular interaction domains bind specifically to each other.
  • 3. A composition comprising:
    • a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and
    • b. a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain;
    • wherein the first-type and third-type biomolecular interaction domains bind specifically to each other.
  • 4. A composition comprising:
    • a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and
    • b. a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; and
    • c. a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain;
    • wherein the second-type and third-type biomolecular interaction domains compete for binding to the first-type biomolecular interaction domain.
  • 5. The composition of any of paragraphs 3-4, wherein the third-type biomolecular interaction domain and first-type biomolecular interaction domain have a higher affinity for each other than the second-type biomolecular interaction domain and first-type biomolecular interaction domain.
  • 6. A composition comprising:
    • a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain;
    • b. a signaling polypeptide comprising a second-type biomolecular interaction domain, a fourth-type biomolecular interaction domain, and an intracellular signaling domain; and
    • c. a recognition polypeptide comprising a second recognition domain and a fifth-type biomolecular interaction domain;
    • wherein the first-type biomolecular interaction domain and the second-type biomolecular interaction domain bind specifically to each other; and
    • wherein the fourth-type biomolecular interaction domain and the fifth-type biomolecular interaction domain bind specifically to each other.
  • 7. The composition of paragraph 6, wherein the fourth-type biomolecular interaction domain and fifth-type biomolecular interaction domain have a weaker affinity than the second-type biomolecular interaction domain and first-type protein interaction domain.
  • 8. The composition of any of paragraphs 6-7, wherein the first polypeptide further comprises a sixth-type biomolecular interaction domain and the recognition polypeptide further comprises a seventh-type biomolecular interaction domain which bind specifically to each other.
  • 9. The composition of any of paragraphs 2-8, wherein the first polypeptide comprises the entire TCR recognition domain.
  • 10. The composition of any of paragraphs 2-8, wherein the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.
  • 11. The composition of any of paragraphs 1-10, wherein the TCR recognition domain comprises a non-polypeptide component.
  • 12. The composition of any of paragraphs 3-11, wherein the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.
  • 13. The composition of any of paragraphs 3-12, wherein the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.
  • 14. The composition of any of the preceding paragraphs, wherein the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; featureless peptide MHC; or a MHC-peptide fusion.
  • 15. The composition of paragraph 14, wherein the peptide is a human or non-human peptide.
  • 16. The composition of any of paragraphs 14-15, wherein the peptide is a Minor Histocompatibility Antigen (MiHA).
  • 17. The composition of any of paragraphs 14-16, wherein the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 18. The composition of any of paragraphs 14-17, wherein the MHC-peptide complex is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 19. The composition of any of paragraphs 14-17, wherein the MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 20. The composition of any of paragraphs 14-17, wherein the MHC is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 21. The composition of any of paragraphs 14-17, wherein the MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 22. The composition of any of paragraphs 14-17, wherein the MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 23. The composition of any of paragraphs 14-22, wherein the MHC is a MHC class I or a MHC class II.
  • 24. The composition of any of paragraphs 1-13, wherein the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion.
  • 25. The composition of paragraph 24, wherein the CD1 is CD1d.
  • 26. The composition of any of the preceding paragraphs, wherein the biomolecular interaction domains are found on an extracellular portion of the respective polypeptides.
  • 27. The composition of any of the preceding paragraphs,
    • a. wherein the biomolecular interaction domain(s) is a leucine zipper, or any binding pair of biomolecular interaction domains are collectively a pair of leucine zippers;
    • b. wherein the biomolecular interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE);
    • c. wherein the biomolecular interaction domain(s) is a PSD95-Dlg1-zo-1 (PDZ) domain;
    • d. wherein the biomolecular interaction domain(s) is a streptavidin and/or a streptavidin binding biomolecular (SBP) or any binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding biomolecular (SBP);
    • e. wherein the biomolecular interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding biomolecular (FKBP);
    • f. wherein the biomolecular interaction domain(s) is a cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506 binding biomolecular (FKBP);
    • g. wherein the biomolecular interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding biomolecular (FKBP);
    • h. wherein the biomolecular interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1);
    • i. wherein the biomolecular interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag;
    • j. wherein the biomolecular interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52);
    • k. wherein the biomolecular interaction domain(s) is a PYL and/or a ABI, or any binding pair of biomolecular interaction domains are collectively a PYL and a ABI;
    • l. wherein the biomolecular interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain;
    • m. wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags;
    • n. wherein the biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding biomolecular or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or
    • o. wherein the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE binding protein.
  • 28. The composition of paragraph 27, wherein the nucleotide tag is a DNA tag or dsDNA tag.
  • 29. The composition of any of the preceding paragraphs, wherein the intracellular signaling domain comprises or is a signaling domain from one or more proteins selected from the group consisting of:
    • TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.
  • 30. A cell comprising and/or expressing the composition of any of the preceding paragraphs.
  • 31. A composition comprising a first polypeptide of any of the preceding paragraphs and a cell expressing or comprising the signaling polypeptide of any of the preceding paragraphs.
  • 32. The cell or composition of any of paragraphs 30-31, wherein the TCR recognition domain comprises a MHC allogeneic, autologous, or xenogeneic to the cell.
  • 33. The cell or composition of any of paragraphs 30-32, wherein the TCR recognition domain comprises a synthetic MHC.
  • 34. The cell or composition of any of paragraphs 30-33, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogeneic to the cell.
  • 35. The cell or composition of any of paragraphs 30-34, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.
  • 36. The cell or composition of any of paragraphs 30-35, wherein the cell is a NK cell, dendritic cell, regulatory T cell, or effector T cell.
  • 37. The cell or composition of any of paragraphs 30-36, wherein the cell is engineered to express one of more of the polypeptide(s) of the composition.
  • 38. The cell or composition of any of paragraphs 30-37, wherein the cell is engineered to express the signaling polypeptide of the composition.
  • 39. The cell or composition of any of paragraphs 30-38, wherein the cell is further engineered to knockout or knockdown the native MHCI/II.
  • 40. The cell or composition of any of paragraphs 30-39, wherein the cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.
  • 41. A chimeric antigen receptor (CAR) comprising:
    • a. an anti-CD127 and/or anti-CD45RO recognition domain;
    • b. an intracellular signaling domain.
  • 42. A composition comprising:
    • a first polypeptide comprising:
    • a. an anti-CD127 and/or anti-CD45RO recognition domain;
    • b. a first-type protein interaction domain; and
    • a second polypeptide comprising:
    • c. a second-type protein interaction domain; and
    • d. an intracellular signaling domain;
    • wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.
  • 43. A composition comprising:
    • a first polypeptide comprising:
    • a. an anti-CD127 recognition domain;
    • b. a first-type protein interaction domain;
    • a second polypeptide comprising:
    • c. an anti-CD45RO recognition domain;
    • d. a fifth-type protein interaction domain; and
    • a third polypeptide comprising:
    • e. a second-type and a fourth-type protein interaction domain; and
    • f. an intracellular signaling domain;
    • wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and
    • wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.
  • 44. A cell comprising the CAR or composition of any of paragraphs 41-43.
  • 45. A method of treating or preventing an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD in a subject in need thereof, the method comprising administering to the subject a composition and/or cell of any of the preceding paragraphs.
  • 46. The method of paragraph 45, wherein the TCR recognition domain comprises a MHC allogeneic to the subject.
  • 47. The method of paragraph 45, wherein the TCR recognition domain comprises a MHC autologous to the transplant cells.
  • 48. The method of paragraph 45, wherein the TCR recognition domain comprises a MHC xenogeneic to the transplant cells.
  • 49. The method of paragraph 45, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic to the subject.
  • 50. The method of paragraph 45, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells.
  • 51. The method of paragraph 45, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells.
  • 52. The method of any of paragraphs 45-51, wherein the MHC and/or the peptide is synthetic.
  • 53. The method of any of paragraphs 45-52, wherein the transplant is any human or non-human cell, tissue, or organ.
  • 54. The method of any of paragraphs 45-53, wherein the transplant is an allogeneic hematopoietic stem cell or solid organ transplantation.
  • 55. The method of any of paragraphs 45-52, wherein the malignant T cell condition is T cell acute lymphoblastic leukemia or T cell lymphoblastic lymphoma.
  • 56. The method of any of paragraphs 45-52, wherein the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma.
  • 57. The method of any of paragraphs 45-52, wherein the autoimmune disease is thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis), palindromic arthritis, inflammatory hyperproliferative skin diseases, psoriasis such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and psoriasis of the nails, atopy including atopic diseases such as hay fever and Job's syndrome, dermatitis including contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, non-specific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic intraocular inflammatory diseases, urticaria such as chronic allergic urticaria and chronic idiopathic urticaria, including chronic autoimmune urticaria, myositis, polymyositis/dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic scleroderma), sclerosis such as systemic sclerosis, multiple sclerosis (MS) such as spino-optical MS, primary progressive MS (PPMS), and relapsing remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, sclerosis disseminata, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (for example, Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis such as ulcerative colitis, colitis ulcerosa, microscopic colitis, collagenous colitis, colitis polyposa, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), bowel inflammation, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome, including adult or acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, an autoimmune hematological disorder, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as in meningitis, herpes gestationis, pemphigoid gestationis, pruritis scroti, autoimmune premature ovarian failure, sudden hearing loss due to an autoimmune condition, IgE-mediated diseases such as anaphylaxis and allergic and atopic rhinitis, encephalitis such as Rasmussen's encephalitis and limbic and/or brainstem encephalitis, uveitis, such as anterior uveitis, acute anterior uveitis, granulomatous uveitis, nongranulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome such as chronic or acute glomerulonephritis such as primary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membrano- or membranous proliferative GN (MPGN), including Type I and Type II, and rapidly progressive GN, proliferative nephritis, autoimmune polyglandular endocrine failure, balanitis including balanitis circumscripta plasmacellularis, balanoposthitis, erythema annulare centrifugum, erythema dyschromicum perstans, eythema multiform, granuloma annulare, lichen nitidus, lichen sclerosus et atrophicus, lichen simplex chronicus, lichen spinulosus, lichen planus, lamellar ichthyosis, epidermolytic hyperkeratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, allergic reaction, eczema including allergic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and vesicular palmoplantar eczema, asthma such as asthma bronchiale, bronchial asthma, and auto-immune asthma, conditions involving infiltration of T cells and chronic inflammatory responses, immune reactions against foreign antigens such as fetal A-B-O blood groups during pregnancy, chronic pulmonary inflammatory disease, autoimmune myocarditis, leukocyte adhesion deficiency, lupus, including lupus nephritis, lupus cerebritis, pediatric lupus, non-renal lupus, extra-renal lupus, discoid lupus and discoid lupus erythematosus, alopecia lupus, systemic lupus erythematosus (SLE) such as cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and lupus erythematosus disseminatus, juvenile onset (Type I) diabetes mellitus, including pediatric insulin-dependent diabetes mellitus (IDDM), adult onset diabetes mellitus (Type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic large-artery disorder, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, sarcoidosis, granulomatosis including lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitides, including vasculitis, large-vessel vasculitis (including polymyalgia rheumatica and giant-cell (Takayasu's) arteritis), medium-vessel vasculitis (including Kawasaki's disease and polyarteritis nodosa/periarteritis nodosa), microscopic polyarteritis, immunovasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, necrotizing vasculitis such as systemic necrotizing vasculitis, and ANCA-associated vasculitis, such as Churg-Strauss vasculitis or syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs positive anemia, Diamond Blackfan anemia, hemolytic anemia or immune hemolytic anemia including autoimmune hemolytic anemia (AIHA), pernicious anemia (anemia perniciosa), Addison's disease, pure red cell anemia or aplasia (PRCA), Factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases involving leukocyte diapedesis, CNS inflammatory disorders, multiple organ injury syndrome such as those secondary to septicemia, trauma or hemorrhage, antigen-antibody complex-mediated diseases, anti-glomerular basement membrane disease, anti-phospholipid antibody syndrome, allergic neuritis, Behcet's disease/syndrome, Castleman's syndrome, Goodpasture's syndrome, Reynaud's syndrome, Sjogren's syndrome, Stevens-Johnson syndrome, pemphigoid such as pemphigoid bullous and skin pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, pemphigus mucus-membrane pemphigoid, and pemphigus erythematosus), autoimmune polyendocrinopathies, Reiter's disease or syndrome, an immune complex disorder such as immune complex nephritis, antibody-mediated nephritis, polyneuropathies, chronic neuropathy such as IgM polyneuropathies or IgM-mediated neuropathy, and autoimmune or immune-mediated thrombocytopenia such as idiopathic thrombocytopenic purpura (ITP) including chronic or acute ITP, scleritis such as idiopathic cerato-scleritis, episcleritis, autoimmune disease of the testis and ovary including autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis such as autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis), or subacute thyroiditis, idiopathic hypothyroidism, Grave's disease, polyglandular syndromes such as autoimmune polyglandular syndromes (or polyglandular endocrinopathy syndromes), paraneoplastic syndromes, including neurologic paraneoplastic syndromes such as Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis such as allergic encephalomyelitis or encephalomyelitis allergica and experimental allergic encephalomyelitis (EAE), myasthenia gravis such as thymoma-associated myasthenia gravis, cerebellar degeneration, neuromyotonia, opsoclonus or opsoclonus myoclonus syndrome (OMS), and sensory neuropathy, multifocal motor neuropathy, Sheehan's syndrome, autoimmune hepatitis, lupoid hepatitis, giant-cell hepatitis, autoimmune chronic active hepatitis, lymphoid interstitial pneumonitis (LIP), bronchiolitis obliterans (non-transplant) vs NSIP, Guillain-Barre syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis such as primary biliary cirrhosis and pneumonocirrhosis, autoimmune enteropathy syndrome, Celiac or Coeliac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, amylotrophic lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear disease such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsed or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome/nonsyphilitic interstitial keratitis, Bell's palsy, Sweet's disease/syndrome, rosacea autoimmune, zoster-associated pain, amyloidosis, a non-cancerous lymphocytosis, a primary lymphocytosis, which includes monoclonal B cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathy, paraneoplastic syndrome, channelopathies including channelopathies of the CNS, autism, inflammatory myopathy, focal or segmental or focal segmental glomerulosclerosis (FSGS), endocrine opthalmopathy, uveoretinitis, chorioretinitis, autoimmune hepatological disorder, fibromyalgia, multiple endocrine failure, Schmidt's syndrome, adrenalitis, gastric atrophy, presenile dementia, demyelinating diseases such as autoimmune demyelinating diseases and chronic inflammatory demyelinating polyneuropathy, Dressler's syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility, e.g., due to anti-spermatozoan antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortion, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, bird-fancier's lung, allergic granulomatous angiitis, benign lymphocytic angiitis, Alport's syndrome, alveolitis such as allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reaction, Sampter's syndrome, Caplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial lung fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum et diutinum, erythroblastosis fetalis, eosinophilic faciitis, Shulman's syndrome, Felty's syndrome, cyclitis such as chronic cyclitis, heterochronic cyclitis, iridocyclitis (acute or chronic), or Fuch's cyclitis, Henoch-Schonlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndromes, Evan's syndrome, autoimmune gonadal failure, Sydenham's chorea, post-streptococcal nephritis, thromboangitis ubiterans, thyrotoxicosis, tabes dorsalis, chorioiditis, giant-cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplant organ reperfusion, retinal autoimmunity, aphthae, aphthous stomatitis, arteriosclerotic disorders, aspermiogenesis, autoimmune hemolysis, Boeck's disease, enteritis allergica, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, febris rheumatica, Hamman-Rich's disease, sensoneural hearing loss, ileitis regionalis, leucopenia, transverse myelitis, primary idiopathic myxedema, ophthalmia symphatica, polyradiculitis acuta, pyoderma gangrenosum, acquired spenic atrophy, vitiligo, toxic-shock syndrome, conditions involving infiltration of T cells, leukocyte-adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, diseases involving leukocyte diapedesis, multiple organ injury syndrome, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathies, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine failure, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid, frontal, maxillary, or sphenoid sinusitis, an eosinophil-related disorder such as eosinophilia, pulmonary infiltration eosinophilia, eosinophilia-myalgia syndrome, Loffler's syndrome, chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, granulomas containing eosinophils, seronegative spondyloarthritides, polyendocrine autoimmune disease, sclerosing cholangitis, sclera, episclera, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia telangiectasia syndrome, angiectasis, autoimmune disorders associated with collagen disease, rheumatism, allergic hypersensitivity disorders, glomerulonephritides, reperfusion injury, ischemic re-perfusion disorder, lymphomatous tracheobronchitis, inflammatory dermatoses, dermatoses with acute inflammatory components, and autoimmune uveoretinitis (AUR).
  • 58. The method of any of paragraphs 45-52, wherein the T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy.
  • 59. The method of paragraph 58, wherein the biologic, cell-therapy, or gene therapy is an adeno-associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.
  • 60. The method of any of paragraphs 45-52, wherein the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129.
  • 61. The method of any of paragraphs 45-52, wherein the disease is vitiligo and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO: 2021.
  • 62. The method of any of paragraphs 45-52, wherein the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO: 2039.
  • 63. The method of any of paragraphs 45-52, wherein the disease is type 1 diabetes and the TCR recognition domain comprises one or more of SEQ ID NOs: 8-17; comprises HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or comprises HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129.
  • 64. The method of any of paragraphs 45-52, wherein the disease is vitiligo and the TCR recognition domain comprises SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or comprises HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or comprises HLA-A*0101 and SEQ ID NO: 2021.
  • 65. The method of any of paragraphs 45-52, wherein the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or comprises HLA-B*0702 and SEQ ID NO: 2038; or comprises HLA-B*0801 and SEQ ID NO: 2039.

Some embodiments of the technology described herein can be defined according to any of the following numbered paragraphs:

• • 1. A composition, comprising:
    • a. a TCR recognition domain; and
    • one or both of:
    • b. an intracellular signaling domain; and
    • c. a biomolecular interaction domain.
  • 2. The composition of paragraph 1, comprising a TCR recognition domain and a biomolecular interaction domain.
  • 3. The composition of paragraph 1, comprising a TCR recognition domain and an intracellular signaling domain.
  • 4. The composition of paragraph 1, wherein the biomolecular interaction domain of c) is a first-type biomolecular interaction domain and the composition further comprises a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; wherein the first-type and second-type biomolecular interaction domains bind specifically to each other.
  • 5. The composition of paragraph 1, wherein the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; a featureless peptide MHC; or a MHC-peptide fusion.
  • 6. The composition of paragraph 1, wherein the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion.
  • 7. The composition of paragraph 6, wherein the CD1 is CD1d.
  • 8. The composition of paragraph 1, wherein the TCR recognition domain is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
  • 9. The composition of paragraph 1,
    • a. wherein the biomolecular interaction domain(s) is a leucine zipper, or any binding pair of biomolecular interaction domains are collectively a pair of leucine zippers;
    • b. wherein the biomolecular interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE);
    • c. wherein the biomolecular interaction domain(s) is a PSD95-Dlg1-zo-1 (PDZ) domain;
    • d. wherein the biomolecular interaction domain(s) is a streptavidin and/or a streptavidin binding biomolecular (SBP) or any binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding biomolecular (SBP);
    • e. wherein the biomolecular interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding biomolecular (FKBP);
    • f. wherein the biomolecular interaction domain(s) is a cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506 binding biomolecular (FKBP);
    • g. wherein the biomolecular interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding biomolecular (FKBP) or any binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding biomolecular (FKBP);
    • h. wherein the biomolecular interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1);
    • i. wherein the biomolecular interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag;
    • j. wherein the biomolecular interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52);
    • k. wherein the biomolecular interaction domain(s) is a PYL and/or a ABI, or any binding pair of biomolecular interaction domains are collectively a PYL and a ABI;
    • l. wherein the biomolecular interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain;
    • m. wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags;
    • n. wherein the biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding biomolecular or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or
    • o. wherein the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE binding protein.
  • 10. The composition of paragraph 9, wherein the nucleotide tag is a DNA tag or dsDNA tag.
  • 11. The composition of paragraph 2, further comprising a cell expressing or comprising the signaling polypeptide.
  • 12. The composition of paragraph 11, wherein the TCR recognition domain is allogeneic, autologous, or xenogenic to the cell.
  • 13. The composition of paragraph 11, wherein the TCR recognition domain is synthetic.
  • 14. The composition of paragraph 11, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogenic to the cell.
  • 15. The composition of paragraph 11, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.
  • 16. The composition of paragraph 11, wherein the cell is a NK cell, dendritic cell, regulatory T cell, or effector T cell.
  • 17. The composition of paragraph 11, wherein the cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.
  • 18. A method of treating or preventing an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD in a subject in need thereof, the method comprising administering to the subject a composition of paragraph 1.
  • 19. The method of paragraph 18, wherein the transplant is an allogeneic hematopoietic stem cell or solid organ transplantation.
  • 20. The method of paragraph 18, wherein the malignant T cell condition is T cell acute lymphoblastic leukemia or T cell lymphoblastic lymphoma.
  • 21. The method of paragraph 18, wherein the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma.
  • 22. The method of paragraph 18, wherein the T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy.
  • 23. The method of paragraph 22, wherein the biologic, cell-therapy, or gene therapy is an adeno-associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.
  • 24. The method of paragraph 18, wherein the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129.
  • 25. The method of paragraph 18, wherein the disease is vitiligo and the TCR recognition domain comprises sequences with at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO: 2021.
  • 26. The method of paragraph 18, wherein the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO: 2039.

EXAMPLES

Example 1

Auto- and allo-reactive T cells attacking patient's or donor's cells or organs is the major cause of autoimmunity and transplant rejection. Current treatments involve stringent immunosuppressant therapy, which can lead to severe side effects. Specifically, depletion of auto and alloreactive T cells prevents autoimmunity and transplant rejection without immunosuppressant modalities. T cells engineered with a Chimeric antigen ligand (CAL) can redirect their specificity toward the pathologic T cells. T cells engineered with a Chimeric antigen receptor (CAR) can redirect their specificity and have already been approved to treat some types of B cell malignancies. Currently, engineered regulatory T cell, which can inhibit immune reactions in an antigen-dependent manner, is under investigation to expand the application of CAR T cells therapies such as autoimmune disease and transplant rejection.

To specifically deplete disease causing T cells, described herein is a CAL and/or CAR that can recognize and bind to the specific T cell receptor on the disease causing T cells. This CAL and/or CAR is a composed of a peptide-HLA (e.g., a monomer or multimer or oligomer thereof) as the recognition domain fused to signaling domains from T cell receptors. A split version of the system can also be generated where the system is composed of two pieces. One piece is an universal CAR with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain. The second piece is an adaptor molecule, e.g, a CAL composed of a peptide-HLA oligomers (or monomer or multimer) fused to the cognate biomolecular interaction domain.

Most CAR T cells therapies for autoimmune disease and transplant rejection uses regulatory T cells and are designed to target the organ to provide organ-specific, e.g., local immunosuppression. In contrast, the present, e.g., CAL, design is targeting the source immune cells inside that patient that are causing the disease. Accordingly, provided herein is a cell-based therapy for autoimmune diseases and transplant rejection.

Example 2

Dendritic cells (DCs) are the interconnection between innate and adaptive immune system and are able to induce protective immune responses following stimulation by a variety of stimuli. Based on their phenotype and function, DCs can be divided into conventional DCs and plasmacytoid DCs (pDCs). Many studies have shown that DCs are essential mediators of pro-inflammatory or anti-inflammatory (tolerogenic) responses. The subsets of DCs that suppress immune responses are generally known as tolerogenic DCs because of their functions in inducing T cell apoptosis, anergy and regulatory T cells (Tregs).

A tolerogenic state in DCs (tol-DC) can be induced using several pharmacological agents, such as cyclosporine A, rapamycin, dexamethasone, vitamin A, vitamin D or other cytokines and growth factors. Recently, the insertion of exogenous DNA to enhance tol-DC function has been investigated as a therapeutic possibility for treating autoimmune diseases. ‘Killer’ DCs, obtained by transfection with DNA encoding FasL or TNF-related apoptosis-inducing ligand, efficiently induce T-cell apoptosis and prevent the rejection of heart grafts in animal models.

However, the effect of these DCs are not specific. Overexpression of inhibitory molecules, including IL-10, TGF-β, CTLA-4 and SOCS1, enable tol-DCs to more efficiently induce Tregs which might cause systemic immune suppression.

Alloreactive immune cells can be suppressed in a specific manner using the methods described herein, e.g., by using a universal UniCAR DC system that presents the donors' pMHC tetra/dextramers (e.g., the CAL) and binds to a genetically engineered DC (K/O MHC I/II but presents donor peptide on the tetra/dextramer). In some embodiments, the pMHC can be provided as a monomer, oligomer, or multimer.

Any form of the recipient pMHC (e.g., monomer/oligomer/tetramer/dextramer)+targeting module (e.g, the CAL) can be used with CAR-DC for the suppression of allo-reactive T-cells in the recipients. A library of identified different alloantigens+correlated or associated with MHC monomers/oligomers/tetramers/dextramers (e.g. HLA-A2+insulin peptide) and attached to the targeting molecules of the CAL and/or CAR system can be produced. Such a library can be commercially available to be combined with DCs for targeting and destroying the auto reactive T-cells.

Cells can be produced that are genetically modified with the specific gene deletion of some genes (IL-12 and NF-κB, MHC I and MHC II [The targeting module will provide these cells with the donor's MHCs]) and insertion of some other genes (IL-10, TGF-β, CTLA-4 and SOCS1). These cells can be commercially ready to use and could be combined with the specific donor, pMHC and targeting module chosen from the aforementioned library.

Example 3

Alloreactive memory T cells have been shown to have the key role in the activation of the recipient immune system and rejection of the allograft. They are known to be the main barrier for tolerance induction through mixed chimerism and other strategies. These cells are very resilient and not responsive to the preconditioning protocols and they recover from irradiation and T cell depletion rapidly. By using CAR-T-effector (Conventional, Supra, Universal CAL or CAR) and attaching any form of the donors' pMHC (monomer, oligomer, tetramer or dextramer) (e.g., a CAL) to the system, the resulting compositions of the invention can be used to decrease or eliminate the need for improved immunosuppressant therapy in the context of transplantation. The engraftment of hematopoietic stem cells can achieve durable mixed chimerism with minimal or no need for toxic preconditioning protocols.

If donor pMHC (monomer/oligomer/tetramer/dextramer; e.g., a CAL) is used in combination with CAR T effector for the abrogation of the allo/xeno reactive T-cells in the recipient, the need for immunosuppressants would be decreased or eliminated. A library of different Donor MHC monomers/oligomers/tetramers attached to the targeting molecules (e.g., the CALs) of the SUPRA/UNI/universal CAL and/or CAR system can be produced to be commercially available. A shelf ready library of the most prevalent MHC (+immunologic wildtype or synthetic peptide molecules that could be used to be combined with the MHC) monomers/oligomers/tetramers could be commercially produced and offered to be used with this system.

Example 4

The involvement of the adaptive immune system in auto-immune diseases has been extensively characterized. T cells are critical contributors to autoimmune diseases. Conventional T (Teff or T helper) cell subsets that play a role in B cell activation and differentiation produce various inflammatory cytokines and destroy target cells with direct cytotoxicity. CAR-T cells have been used to destroy autoimmune B and T cells in a fashion similar to the way in which CD19CAR T cells target and destroy leukemia cells. Targeting autoreactive memory T and B-cells has shown some results. As these methods are nonspecific, they induce some extent of generalized immune suppression and they are not completely effective. The correlation of particular peptide+MHC molecules with certain autoimmune conditions have been expansively studied. By using the available human/animal model pMHC monomer/oligomer/tetra/dextramers and combining them with the SUPRA/UNI/universal CAL and/or CAR technology, we are able to target the autoreactive immune cells in a highly specific manner.

If any form of auto-antigen on pMHC (monomer/oligomer/tetramer/dextramer) (e.g., the CAL) is used in combination with CAR T effector/Treg to abrogate autoreactive T-cells in the recipient, the need for immunomodulatory drugs would be decreased or eliminated. A library of identified autoantigens and correlated MHC monomer/oligomer/tetra/dextramers (e.g. HLA-A2+insulin peptide) attached to the targeting molecules of the SUPRA/UNI/universal CAL and/or CAR system can be produced to be commercially available to be combined with ideal SUPRA or universal CAR T-reg/T effector for targeting and destroying the autoreactive T-cells.

Example 5

Although new advances have increased survival after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) is still the leading cause of late morbidity and mortality after transplant. Current treatment choices are limited in efficacy specifically in steroid-refractory disease, and there is no robust data to help with management decisions.

Adoptive T cell therapy (ACT) refers to the therapeutic use of T cells. T cells genetically engineered to express chimeric antigen receptors (CAR) constitute the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemias and lymphomas and have produced remarkable results in the clinic. The technology described herein permits the opportunity to target diseased cells with specific antigens or receptors very accurately. In the context of GvHD, universal UNICAR T cells can be designed to find the recipient reactive T cell in the donor T cell populations. If the pMHC of the recipients is fused to a Target Module that binds to the UniCAR (e.g, to form a CAL), this system can recognize the TCR repertoires in the donor T/B cell population that can bind to those MHCs. If the recipients' pMHCs are fused to a target module that binds to the universal CAR, this system can recognize and bind to the T cell repertoires in the donor T cell population that can bind to those MHCs.

If the recipient pMHC (monomer/oligomer/tetramer/dextramer) (e.g., a CAL) is used in combination with CAR T effector/Treg for the abrogation of the recipient reactive T-cells in the donor HSC, these reactive T-cells will be depleted and GvHD would not happen. A library of identified different antigens+correlated human/animal models MHC tetramers/dextramers (e.g. HLA-A2+ peptide) attached to the targeting molecules of the SUPRA/UNI CAR system can be produced to be commercially being available to be combined with ideal SUPRA CAR T-reg/T effector for targeting and destroying the reactive T-cells against the recipients. A library of wild type and/or synthetic MHC monomers/oligomers (e.g. HLA-A2+ peptide) attached to the targeting molecules of the universal CAL system can be generated to be mixed with universal CAL T-reg/T effector for targeting and killing the reactive T-cells against the recipients within the donor T cell population.

Example 6

Although new advances have increased survival after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) is still the leading cause of late morbidity and mortality after transplant. Current treatment choices are limited in efficacy specifically in steroid-refractory disease, and there is no robust data to help with management decisions.

Adoptive T cell therapy (ACT) refers to the therapeutic use of T cells. T cells genetically engineered to express chimeric antigen receptors (CAR) constitute the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemias and lymphomas and produced remarkable results in clinical. This technology provides the opportunity to target the cells with specific antigens or receptors very accurately. In the context of GvHD, UNICAR T cells can be designed to find the recipient reactive T cell in the donor T cell populations. If the pMHC of the recipients be fused to a Target Module that bind to the UniCAR, this system can recognize the TCR repertoires in the donor TB cell population that can bind to those MHCs.

If the recipient pMHC (tetramer/dextramer) is used in combination with CAR T effector/Treg for the abrogation of the recipient reactive T-cells in the donor HSC, these cells will be depleted and GvHD would not happen. A library of identified different antigens+correlated human/animal models MHC tetramers/dextramers (e.g. HLA-A2+peptide) attached to the targeting molecules of the SUPRA/UNI CAR system can be produced to be commercially being available to be combined with ideal SUPRA CAR T-reg/T effector for targeting and destroying the reactive T-cells against the recipients.

Example 7—Engineered Lymphocytes for Prevention of Pediatrics Vascularized Composite Allograft Rejection Engineered

Described herein is the development of a clinically applicable tolerance-inducing regimen for VCA transplantation through the establishment of stable mixed chimerism, augmented by the state-of-the-art CAL and/or CAR T cell adoptive immunotherapy and synthetic biology.

Mixed chimerism in animal models and human allograft recipients have only shown to be transient, suggesting that tolerance here relies on the peripheral inactivation of donor-specific T cell. The difficulty in achieving durable mixed chimerism and long-term graft acceptance is the presence of high levels of alloreactive memory T cells that are known to hinder tolerance induction in sensitized rodents, NHPs, and human. The present approach to achieving stable mixed chimerism is to utilize advanced engineering techniques to generate CAL and/or CAR T cell therapeutics to specifically delete the recipient's alloreactive memory immune cells that are reactive against the donor bone marrow and tissue cells to help with engraftment of HSCT and stable mixed chimerism. Selective depletion of memory T lymphocytes with CAL and/or CAR-T cell therapy can help to achieve durable mixed chimerism and tolerance which eventually leads to an immunosuppressant free regimen. Currently this challenge is addressed by administering several immunosuppressant combinations in combination with intense whole-body irradiation which result in depletion of all the immune cells as a consequence of nonspecific targeting. This shotgun approach imposes a severe immune compromised state to the recipients which subsequently brings myriads of consequences including opportunistic infections and malignancies. The technology herein can delete specifically memory T cells (Aim 1) or only alloreactive memory T cells (Aim 2):

Aim 1: It has been shown that CD127+/CD45RO+ memory T cells have a significant role as central, effector and stem cell memory T cells and are known as the most potent constituents of the alloreactive T cell repertoire. These cells have been shown to be the major contributor in chronic rejection of the allograft. To address the challenge of memory T cells a CAL and/or CAR T-effector cell can be generated with scfv against two general markers of memory T cells (CD127+ and CD45RO+). This will be followed by the induction of mixed chimerism protocol in a double humanized mouse model. The results from this aim will show depletion of memory T-cells increases the efficacy of mixed chimerism protocol by improving the engraftment of hematopoietic stem cells hence helping to eliminate or decrease the need for immunosuppressant in mixed chimerism induction protocols.

Aim 2: Due to the diversity of antigen-specific T cells in the context of transplantation, a CAL and/or CAR system is provided that has the flexibility to locate and attack different alloreactive T cells simultaneously. CAL and/or CAR T-eff cells with donor pMHC can target alloreactive T-cells that have TCR against the donor MHCs. We will start with single and double specific antigen-MHC systems. Then, we will utilize a peptide library that is generated from the donor's allograft peptidome to be loaded on commercially-available, exchangeable peptide-MHC multimers. The combination of these pMHCs and CAL and/or CAR T cells will be used to target and destroy alloreactive T cells of the recipients. This development provides a flexible CAL and/or CAR design that can target antigen specific alloreactive T cells.

Described herein is the investigation of a novel adaptive immune cell therapy strategy, making use of the CAL and/or CAR T-cell technology, for VCA tolerance induction.

Example 8—Engineered Lymphocytes for Prevention of Pediatrics Vascularized Composite Allograft Rejection

Significance: There are nearly 2 million people living with limb loss in the US, where over 185,000 amputations occur each year, most of which are sustained by victims of burn injuries, traffic accidents, and medical conditions¹. 112600 children with amputations were treated in US emergency departments alone from 1990 to 2002. Children are frequently the victims of severe burns with limb loss² and they will suffer from its consequences for the rest of their lives. Over the last decade, vascularized composite allotransplantation (VCA), the transplantation of limbs and face from a deceased donor, has become a good alternative for the reconstruction of devastating injuries of these specialized tissues. VCA represents a unique new treatment option for severe soft tissue defects following burn injury to achieve both psychosocial and functional rehabilitation³. The world's first pediatric bilateral hand transplant was successfully performed between an unrelated donor-recipient pair in 2016⁴. The shortcoming is that despite the use of potent immunosuppressive drugs, acute rejection of “foreign” VCA occurs in up to 90% of patients⁵. Increased doses of immunosuppression, with numerous life-threatening complications, is the current approach to prevent loss of the VCA⁵.

During the course of evolution, our immune system has gained the ability to recognize self from nonself cells and attack the “foreigners” by cells such as T Lymphocytes. The Human Leukocyte Antigen (HLA) system (Major Histocompatibility Complex [MHC] in human) comprises cell surface molecules specialized to present antigenic peptides to the T-cell⁶⁷. These peptides are known as Minor Histocompatibility Antigens (MiHA). MiHA sequences can differ among individuals and many of these differences can be recognized by T cells, hence causing the initiation of the rejection process. T-cells are trained to recognize self from non-self MHC/peptides. Each person can present a combination of 12 different MHC class I/II on the surface of his/her cells. (HLA) typing is used to match recipients and donors MHCs for transplants. A close match between a donor's and a recipient's HLA markers is essential for a successful transplant outcome. A potential donor must match a minimum of 6-8 HLA markers which makes finding an ideal donor very difficult. Even after finding the semi ideal donor these patients will be on immunosuppressants for the rest of their life⁸. While these drugs are generally effective, the sequelae of such chronic immunosuppression are well known, and most recipients continue to develop myriad side effects and complications, including opportunistic infections, multiple organ dysfunction, and malignancies.

Transplantation tolerance that allows for the elimination of immunosuppressive drugs, has been the “holy grail” for transplantation medicine since its beginnings⁹. Developing a reproducible, safe tolerance induction protocol would expand VCA use in burn patients as well as in congenital anomalies as demonstrated by the successful transplantation of the arm and hand⁶ and the lower extremity¹⁰. The extension into other organ transplants is also a clear direction to broaden the significance. The overall goal of this research is to enable the clinically applicable tolerance-inducing regimen for VCA transplantation through the establishment of stable mixed chimerism, augmented by the state-of-the-art adoptive immunotherapy and synthetic biology.

Tolerance can be induced through the development of a mixed chimerism protocol for VCA transplants in partially mismatched subjects^8,11. Mixed chimerism is the use of a donor hematopoietic stem cell transplant (HSCT) along with a VCA to induce a hybrid immune system in the recipient that recognizes the donor VCA as “self” and therefore does not reject it. Mixed chimerism in nonhuman primates (NHPs) and human allograft recipients has only shown to be transient, suggesting that tolerance here relies on the peripheral inactivation of donor-specific T cell. The difficulty in achieving durable mixed chimerism and long-term graft acceptance is the presence of high levels of alloreactive memory T cells that are known to hinder tolerance induction in both sensitized rodents^{12, 13} and NHPs^14,15. The present approach to achieving stable mixed chimerism is to utilize advanced engineering techniques to generate T cell therapeutics to specifically delete the recipient's alloreactive memory immune cells that are reactive against the donor bone marrow and tissue cells.

Cellular Immunotherapy

Recently, T cells that are genetically engineered to express chimeric antigen receptor (CAR) produced remarkable results for the treatment of CD19-positive leukemias and lymphomas, leading to complete remissions in pediatric patients^16,17 and evidencing that immune cells can be targeted and eliminated by CAR T cell technology. Thus far, CAR T therapeutics have led to two FDA-approved therapies—the two medication were licensed by Novartis and Gilead in the value 9 and 12 billion dollars¹⁸. First-generation CARs contained only an extracellular antigen-binding domain, a transmembrane domain, and the signaling domain of CD3z. In later generation CARs, intracellular costimulatory domains, derived from either CD28/4-1BB were added to enhance proliferation, persistence, and activity¹⁹. Recently, to expand the capability of CART cells, we introduced a split, universal, and “programmable” (SUPRA) CAR system that simultaneously encompasses multiple critical features²⁰. This system has the ability to switch targets without reengineering the T cells, finely tune T cell activation strength, and sense and respond to multiple antigens. These features make the split CAR-T technology uniquely suitable for targeting alloreactive T-cells in recipients and enhancing transplantation tolerance.

Mixed Chimerism and Tolerance Induction in VCA

Large animal models of Mixed Chimerism: Our laboratory achieved transient mixed chimerism in two-haplotype full-mismatch MGH miniature swine after musculocutaneous VCA and nonmyeloablative recipient conditioning (total body irradiation (TBI) and transient T-cell depletion) concomitant to bone marrow transplantation (BMT)²¹. While the immunomodulatory effect of BMT was demonstrated through in vitro unresponsiveness to the donor, the chimerism levels fell rapidly following cessation of immunosuppression with resulting VCA rejection and loss. This was the first experimental VCA study to demonstrate that transient chimerism alone is neither sufficient for achieving transplant tolerance nor prolonging VCA survival.

Recently, in order to progress towards clinical translation, the previous Mixed Chimerism Induction Protocols (MCIP) in miniature swine were modified to remove T-cell depletion completely in exchange for a higher dose, TBI and TI commencing two days prior to surgery. Following osteomyocutaneous VCA, a peri-operative course of co-stimulatory blockade was given (on PODO, 2, 4 and 6) in addition to a 30-day regimen of tacrolimus (target levels: 10-15 ng/mL) before gradual taper to discontinuation on POD 45²². With this MCIP, we were indeed able to achieve stable mixed chimerism (n=2) following donor BMT without the development of split tolerance in MHC class I mismatched/class II matched recipients and have successfully withdrawn all immunosuppression for >100 days. Building on these results, the next iteration towards clinical translation of these studies is to decrease the toxicity of current protocol to be able to extend it for achieving stable mixed chimerism across full MHC-mismatch barriers—which can utilize immunomodulatory strategies such as CAL and/or CAR T cell therapy for depletion of alloreactive memory T cells that may eventually result in elimination or decrease in the immunosuppressive regimen for VCA transplantation.

Small animal models of Mixed Chimerism: The principles that underlie tolerance induction through mixed chimerism induction protocol are the same for all species. Briefly, mice can be conditioned with intraperitoneal mAb injections including, anti-CD8 and anti-CD40L, anti-CD154, costim. blockade (on day 0 with respect to the BMT). This will be combined with, TBI to be given 6 h before injection of 2.5 10⁵ bone marrow cells (BMC). The compositions and methods described herein can be used for improving mixed chimerism induction protocol in mice models.

Described herein is a CAL and/or CAR designed to target and destroy alloreactive memory T cells thereby improving the engraftment of hematopoietic stem cells and achieving durable mixed chimerism with minimal or no need for toxic preconditioning protocols. To test this multi-pronged approach, we will first evaluate two different strategies of targeting alloreactive memory T cells. The first approach is to design a CAL and/or CAR system that will specifically attack memory T cells through surface markers that uniquely identify this cell population regardless of their TCR specificity. This approach will spare the other immune cells, and, thus retain a healthy immune system.

A second approach is to design CAL and/or CARs that specifically bind to alloreactive TCRs on the recipient's T cells. To do so, instead of using a single-chain variable fragments (scFvs) that is in all classical CAR T cells, we will use peptide-MHC multimers as the ligand recognition domain on the CAL and/or CAR. However, due to a large number of unique alloreactive TCR, many different CAL and/or CAR/pMHCs are needed. This poses a challenge in designing and introducing these CAL and/or CARs into T cells. To circumvent this challenge, we propose to use a two-component split CAL and/or CAR design composed of universal CAL and/or CAR and an adaptor molecule that bridges the CAL and/or CAR T cells to alloreactive T cells. The universal CAL and/or CAR comprises a scFv targeting FITC fused to TCR signaling domains. The adaptor molecules consist of pMHC tetramer fused to a FITC. The addition of pMHC tetramer will bind to the alloreactive TCR and recruit the universal CAL and/or CAR T cell to kill alloreactive T cells. This design allows us to be able to target many variants of TCR with only one CAL and/or CAR. Next, the donor MHCs will be loaded with donor tissue specific peptides that will be generated by utilizing previously published protocols. We can first design CAL and/or CARs to target mouse MHC in a humanized/skin graft mouse model.

Data-Targeting Alloreactive T-cells with peptide-MHC tetramers

The following have been addressed by in vitro data:

• • 1. Whether targeting a memory T cell by using CAR T+pMHC multimer is obtainable. In this embodiment, the anti-FITC CAR expressed by the Jurkat cells is a CAR and the pMHC multimer is a CAL.
    • A. Would the Tetramer efficiently bind to the CAL and/or CAR and target T cells?
    • B. Would the Jurkat CAL and/or CAR T cells get activated by attaching to the adaptor+Target
    • C. Would the Target OT I cells get activated and try to kill the Killer Target T cells?
  • 2. Whether killing a memory T cell with primary CD8 T cell+pMHC multimer would be feasible?

1. Peptide-MHCs Tetramers Activate Jurkat Cells but not the Target Cells

We used anti-FITC CAR Jurkat cells. These cells are an immortalized line of T lymphocyte cells that are used to study activation, signaling, and expression of various chemokine receptors of T cells. Jurkat cells cannot act as a killer cell but will respond to and get activated by the ligands in the same way as actual T cells. FIG. 1 shows tetramers attaching to and staining OTi target T cells in a dose dependent fashion.

The anti-FTC CAR Jurkat cells used in these experiments comprise the following nucleotide sequence and express the following polypeptide sequence.

DNA sequence of anti-FITC CAR. Nucleotides 1-9 = Kozak sequence.
Nucleotides 10-78 = CD8a leading peptide. Nucleotides 79-831 = anti-FITC ScFv. Nucleotides
832-861 = myc. Nucleotides 874-1008 = CD8 hinge. Nucleotides 1009-1211 = CD28. Nucleotides
1212-1338 = 4-1BB. Nucleotides 1339-1677 = CD3ζ. Nucleotides 1678-2388 mCHERRY.
SEQ ID NO: 2747
gccgccaccATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCC
AGGCCGGCTGCAcaagttcagctcgttgaatccggcggaaaccttgttcaaccggggggttccctccgccttagttgtgccgcatctggttttac
gtttggatcattttccatgtcatgggtgcggcaggctcccggggggggactcgaatgggttgcgggtcttagcgcccgatcaagcctcactcactatgca
gatagcgtaaaaggcaggtttacaatctcacgagacaacgcgaagaactcagtctatcttcagatgaactctctccgagtcgaggatacggctgtctatt
attgtgcacgccgctcctacgattcctcaggctacgccgggcacttttattcatacatggatgtctggggccagggaaccttggtaactgtgtctggagg
aggtggatcagggggtggcggaagtggcggaggtggatcctccgtcttgacccagccctcctcagtcagtgctgcccctggccaaaaggttacaatatca
tgttcaggtagtacgtcaaacataggcaacaactacgtgagttggtaccagcaacatcctggcaaagcacctaagttgatgatctatgatgtcagtaagc
ggccatctggggtacccgaccgattctcaggctcaaagagcggaaactccgcgtccctcgacataagtggcctccaatccgaggacgaggccgactacta
ctgtgctgcttgggacgactctctctcagagtttttgtttgggaccggtacaaaactgactgttcttgggGAACAAAAACTCATCTCAGAA
GAAGATCTGAATGGGGCCGCAACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCA
CCATCGCGTTGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGC
AGTGCACACGAGGGGGCTGGACTTCGCCTGTGATTTTTGGGTGCTGGTGGTGGTTGGTGGAG
TCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGA
GGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGC
AAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAAACGGGGCAG
AAAGAAACTCCTGTATATATTCAAACAACCATTTATGAGACCAGTACAAACTACTCAAGAGG
AAGATGGCTGTAGCTGCCGATTTCCAGAAGAAGAAGAAGGAGGATGTGAACTGAGAGTGAA
GTTCAGCAGGAGCGCAGACGCCCCCGCGTACAAGCAGGGCCAGAACCAGCTCTATAACGAG
CTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTG
AGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGA
AAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCA
AGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTT
CACATGCAGGCCCTGCCCCCTCGCGGCatgGTGAGCAAGGGCGAGGAGGATAACATGGCCAT
CATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTC
GAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAG
GTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGG
CTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCG
AGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCA
GGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCC
CCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGAT
GTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGG
CGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCC
GGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGT
GGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAG
taa
Peptide sequence of anti-FITC CAR. Residues 1-21 = CD8a leading peptide.
Residues 22-274 = anti-FITC ScFv. Residues 275-284 = myc. Residues 289-333 = CD8 hinge. Residues
334-401 = CD28. Residues 402-443 = 4-1BB. Residues 444-556 = CD3ζ. Residues 557-792 mCHERRY.
SEQ ID NO: 2748
MALPVTALLLPLALLLHAARPAAQVQLVESGGNLVQPGGSLRLSCAASGFTFGSFSMSWVRQAP
GGGLEWVAGLSARSSLTHYADSVKGRFTISRDNAKNSVYLQMNSLRVEDTAVYYCARRSYDSS
GYAGHFYSYMDVWGQGTLVTVSGGGGSGGGGSGGGGSSVLTQPSSVSAAPGQKVTISCSGSTS
NIGNNYVSWYQQHPGKAPKLMIYDVSKRPSGVPDRFSGSKSGNSASLDISGLQSEDEADYYCAA
WDDSLSEFLFGTGTKLTVLGEQKLISEEDLNGAATTTPAPRPPTPAPTIALQPLSLRPEACRPAAG
GAVHTRGLDFACDFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPT
RKHYQPYAPPRDFAAYRSKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFS
RSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKM
AEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRGMVSKGEEDNMAIIKEFM
RFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHP
ADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKT
MGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITS
HNEDYTIVEQYERAEGRHSTGGMDELYK

Next, we showed that binding of adaptors to the OTi target T cells would not activate them. To evaluate that, we looked at CD69 expression which is a general activation marker for T cells. Interestingly, although adaptor binds to and stains target T cells (FIG. 1) it did not cause activation of OTi target T cells (FIG. 2).

B. Subsequently, we studied the activation of Jurkat CAR T cells after binding to the adaptors. As shown in FIG. 3, binding of adaptor to the Jurkat T cells causes high expression of CD69 at all measured time points.

This activating binding of Jurkat cells—pMHC is in contrary to the binding of OTi (target)—adaptor (FIG. 2). The best explanation for this interesting finding may lie behind the fact that FITC/anti-FITC binding between Jurkat—pMHC is much stronger than OT I—pMHC as the first one is a high affinity antigen-antibody binding and the second one is a weaker TCR-pMHC binding. This difference in the strength of binding between CAR—adaptor and Target—adaptor makes this novel approach an ideal setting for the purpose of targeting memory T cells.

C. We also looked at the Jurkat cells count after 24 hrs. as an indicator of their condition and whether this activation would cause any cell death indicated by a decrease in their numbers (FIG. 2).

2. Primary CD8 Anti-FITC CAR T Cells can Specifically Target and Kill Alloreactive OTi Cells.

The previous set of experiments with Jurkat+adaptor+OTi target cells confirmed the feasibility of targeting alloreactive T cells by adaptor molecules and showed activation of CAR T cells against the target cells. Next, we wanted to study whether primary CD8 CAR T killer cells+adaptor molecules can be used not only to identify but also to kill the OTi T cells. As shown in the previous set of experiments the binding between pMHC and target cell was not causing activation of target T cells in contrary to the adaptor (FITC+pMHC, e.g., the CAL)—CAR T cells binding. Hence, we used human primary CD8 T cells to see whether the strength of CART+adaptor (e.g, CAL)+target is enough to initiate and accomplish the killing process. Different concentrations of adaptor+CD8 CART were co-cultured with OTi T cell to see the activation and cytotoxicity of each T cell type. As shown in FIGS. 4 and 5, CD8 CAR T cells cytotoxicity on OTi cells is dependent on the adaptor (e.g., CAL) concentration and was over to 50% with the maximum dose that we used. Additionally, this cytotoxicity is highly specific. (FIG. 5) Human CD8 anti-FITC CART cells were used here. Similar validations can be conducted with mouse anti-FITC CARs in both in vivo and in vitro experiments.

Described herein is a novel method to use pMHC multimers for specifically targeting and destroying allo/autoreactive T cells responsible for graft rejection and autoimmune diseases. Our approach is the first demonstration of utilizing pMHC multimer (e.g, CAL) in combination with CAR T cell technology for specific targeting of alloreactive T cell population. We believe this approach is the most specific strategy for eliminating the destructive role of alloreactive T cells in transplantation. The potential application of this method extends far beyond elimination of immunosuppressants in transplantation rejection and GVHD treatment and could be applicable to many autoimmune conditions including type 1 diabetes, multiple sclerosis, rheumatoid arthritis, scleroderma, and myriad disorders that originate from auto reactive T cells dysfunction.

Provided herein is a novel approach for targeting the memory repertoire with anti CD45RO/CD127 with CAL and/or CAR T cell therapy. Although this strategy might not be as specific as using pMHC multimers, it still conveys a significant novelty. By depleting the memory T cells population there is much less of a need for induction protocols.

Also contemplated herein are individualized peptide libraries based on the differences between the recipient and the donor proteome and immune-peptidome. Hundreds to thousands of MHC-associated peptides can now be identified in a single measurement using optimal biological model systems. As mentioned earlier, studies involving MHC identical grafts indicate that minor histocompatibility antigens may also mediate rejection. Human Immuno-Peptidome Project (HIPP) is an international project that was created to accelerate research toward robust and comprehensive analysis of immunopeptidome. HIPP has published the technical guidelines that represent the information required to sufficiently conduct and interpret all of the immunopeptidomics experiments. These nonhomogeneous peptides can potentially initiate rejection through activating alloreactive T cells.

By utilizing these peptides in combination with exchangeable MHC tetramer systems, we can employ these peptides to identify and target donor reactive T cells. The default peptide on exchangeable peptide MHC systems can be removed by UV light exposure and replaced by any desired peptide. The donor multimers MHCs can be generated using commercially available services, unloaded with UV and then loaded with donor tissue specific peptides that will be generated by utilizing previously published protocols. By utilizing these peptides in combination with exchangeable MHC tetramer systems, we can employ these peptides to identify and target donor reactive T cells.

As described herein, selective depletion of memory T cells with CAR-T cell therapy can help in achieving durable mixed chimerism and tolerance which lead to the elimination or decrease in immunosuppressive regimens. Currently this challenge is addressed by administering intense whole-body irradiation with several immunosuppressant combinations for targeting all the immune cell populations. This shotgun approach results in severe immune compromised state in the recipient which subsequently brings myriads of side effects and complications, including opportunistic infections, reno-vascular dysfunction, and malignancies.

Memory T cells (CD127+/CD45RO+) are known to be the main contributor in rejecting allografts 2425. Hence, their depletion is expected to help engraftment of HSC and increase the survival of allografts. To investigate this hypothesis, we will generate CAL and/or CAR T-effector cell with scfv against a general marker of memory T cells (CD127+ and CD45RO+). This will be followed by induction of mixed chimerism protocol in a humanized mice model. The engraftment and stability of mixed chimerism will be followed by the presence of donor lymphoid and myeloid cells in the recipient blood stream until the study endpoints.

Activity 1.1 Generation and evaluation of AND logic CAL and/or CAR T cells to only target cells co-expressing both CD127+ and CD45RO+. Since no single surface marker can uniquely specify memory T cells, we will need to target a combination of CD markers to ensure specificity. We chose CD127 and CD45RO because they have proven to uniquely identify memory T cells 2627. To accomplish this combinatorial targeting, we will leverage an advance CAR system called SUPRA. This (SUPRA) CAR is a two-component receptor system comprises a universal receptor (zipCAR) expressed on T cells and a targeting scFv adaptor (zipFv). The zipCAR universal receptor is generated from the fusion of intracellular signaling domains (FIG. 6) and a leucine zipper as the extracellular domain. The zipFv adaptor molecule is generated from the fusion of a cognate leucine zipper and an scFv. The scFv on the zipFv binds to the antigen, while the leucine zipper binds to and activates the zipCAL and/or zipCAR on the T cells. For transplantation immunotherapy, these features can mitigate over-activation and enhance specificity.

The SUPRA CAL and/or CAR system can also be designed to perform AND combinatorial logic of antigen recognition. Orthogonal leucine zipper pairs can be used to generate CAL and/or CARs with split signaling domains (e.g., CD3z, CD28, 4-1BB), thus enabling independent and simultaneous control of these pathways (FIG. 6B). T cells need both CD3z and costimulatory signaling simultaneously to be fully activated therefore, each CAL and/or CAR can be readily paired with scFvs that target different antigens, thereby enabling two antigen combinatorial and logical antigen sensing.

Using the SUPRA system in human primary T cells, we have demonstrated that the activity of SUPRA CARs can be finely regulated via multiple mechanisms to limit over-activation (FIG. 7). SUPRA CAR T cells can also logically respond to multiple antigens for improved target specificity. Furthermore, orthogonal SUPRA CARs can be used to inducibly regulate multiple signaling pathways or T cell subtypes to increase the breadth of immune responses that can be achieved. A summary of the SUPRA CAL and/or CAR T cell features used for this project can be found in FIGS. 6A-6B, 7A-7D.

zipCAR Receptor Construct Design: As described herein, AND logic CAL and/or CAR T are generated by fusing different leucine zippers to the hinge region of the human CD8a chain and transmembrane and cytoplasmic regions of the human CD28, or CD3z signaling endo-domains. All CAL and/or CARs can contain a myc tag to verify surface expression. Besides, these primary T cells will also be fused to mCherry after CD3z chain to visualize expression.

zipFv Construct Design: The zipFv molecules contain an scFv against CD127 fused to an SYN2 (to stimulate the CD3z SYN1 zipCAR), and an scFv against CD45RO fused to a JUN zipper (to stimulate the costimulatory FOS zipCAR). The scfvs sequences are available and can be constructed rapidly through commercial DNA synthesis²⁸.

zipCAR transduction: Human PBMC are purified using separation kits. are introduced into primary human T cells via retroviral transduction. Expression of zipCARs is quantified via myc and V5-tag immunostaining and flow cytometry. As controls, a zipCAR that contains both CD3z and CD28 (i.e. SYN1-CD28-CD3z) can be used. This zipCAR cannot perform logic computation. A CD45RO-SYN2 or CD127-SYN2 zipFv can be added to ensure that zipCARs can kill CD45RO+ or CD127+ cells. These controls will also serve as reference for specificity.

Evaluate the Activity and Efficacy of Anti-CD127+/CD45RO+CAR T Cells In Vitro

Co-culturing PBMCs with CD127+/CD45RO+ cells with SUPRA CART cells to investigate the specificity of targeting

This experiment examines whether this system specifically kills CD127+ and CD45RO+ double positive T cells amongst a large population of PBMCs. PBMCs are obtained either from known VCA recipients or Mass General Blood Bank Center after being approved by IRB. First, human CD45RO+ and CD127+ are stained to identify the initial percentage of memory cells in the whole population (CD45RO/CD127 double positive vs double neg). Next, PBMCs are co-cultured with zipCAR expressing CD8+ T cells with zipFvs (anti CD127 and anti CD45RO) with the same condition. At the study time points (24, 48, 72 hrs) significant changes in the ratio of CD45RO/CD127+/+ to −/− will be considered as an indicator of whether SUPRA CARs were efficient in targeting specific T cells rather than non-specific cytotoxic effects. Additionally, engineered T cells and zipFvs will be cocultured with the memory T cells at 3 different zipFvs concentrations (5, 25, 50 ng/well) to determine the correlation between cell killing and zipFv concentration. T cell activation (CD69 expression, IL-2 and IFN-g in the media) and cytotoxicity against memory T cells (no. of remaining live cells) will also be measured. The proliferation of T cells will be measured by cell counting. All conditions will be tested at least in triplicate.

TABLE 1
outline of experiments

	1	PBMC-Ctrl
	2	PBMC + SUPRA CAR T
	3	PBMC + SUPRA CAR T + scFv against CD45RO
	4	PBMC + SUPRA CAR T + scFv against CD127
	5	PBMC + SUPRA CAR T + scFvs CD127 + CD45RO

To Evaluate and Characterize the Effect of Memory T Cells on the HSCT Engraftment.

Generation of a Humanized Mouse Model with the VCA-Recipients PBMCs

To examine the effect of CD45RO/CD127 targeting with CAR T cells on the graft take and mixed chimerism sustainability, a humanized mouse model is generated by utilizing the PMBCs of VCA-recipients (FIGS. 8A, 8B). The humanized PBMC model has the fastest engraftment rate using adult PBMCs and enables studies that require a strong memory T cell function. VCA recipients' PBMCs hold an alloreactive population of memory T cells (CD45RO+/CD127+) and makes this a reliable model for studying their depletion effects on the rejection of the graft. The number of mice used per group in this study is chosen based on the recommendations from previous studies²⁹. In brief, irradiated NSG (MHC I/II K/O) (3 Gy) mice will be injected with 5×105PBMC cells and human peripheral reconstitution is measured by flow cytometry every other week³⁰.

Testing the effects of memory T cells depletion on mixed chimerism establishment cells in a skin graft humanized mouse model: The NSG (MHC I/II K/O) mouse model that was generated by using the VCA-recipient PBMCs are used as the foundation for induction of mixed chimerism. The presence of memory T cells in the human PBMCs of this model imitates the challenges for induction of mixed chimerism in human subjects. NSG Hu-PBMCs are treated with/without (ctrl) anti CD45RO/CD127 CAR T to deplete the memory compartment of their T cells population (FIGS. 8A, 8B).

Fetal CD34+ HSC (and skin tissue) can be obtained from Advanced Bioscience Resources. Fetal HSCs are utilized to evaluate the generation and establishment of multi-lineage engraftment by using previously published Mixed Chimerism Induction Protocols (MCIP) in mice 23. In short, male mice are treated with anti CD154, anti-CD40, CTLA-4 and receive 3 Gy TBI. Six hours later they receive 2×105CD34+ HSC bone marrow cells. Engraftment is determined by measuring the presence of donor myeloid and lymphoid cells at different time points. Mixed chimerism is defined as at least 5% of leukocytes in each of the lineages being donor derived. Less than 5% or loss of multi-lineage status is considered as failure of chimerism.

Next, the aforementioned humanized mouse model (VCA-Recipient PBMCs-CD34 HSC) is utilized as a skin graft model to test the effect of depleting CD45RO/CD127 cells on allograft survival. Fetal skin graft is obtained from Advanced Bioscience Resources from the same CD34+HSC donor. At week 10 post PBMC injection, skin grafts from the HSC donor and another allogeneic source are used to test whether nonspecific depletion of memory T cell repertoire affects the skin grafts survival (FIG. 8, 9). Mice are followed until graft rejection or POD 100.

Skin grafting will be conducted using previously published protocols 31. In short, 2 pieces of tail skin from the donor are transplanted on the back of one recipient. Following dressing removal (POD7), mice and their skin grafts are evaluated daily. Graft failure is determined by the physical appearance of the graft; including color, texture, and contour. Histology is also performed to provide supportive data regarding the clinical signs of graft rejection or engraftment. Grafts with more than 50% shrinkage in size or change in color and flexibility (dark and firm grafts) are considered as failed. Skin graft are followed for 100 days.

TABLE 2
outline of experiments

		Recipient (NSG-Hu-VCA-		Skin
#	N	Rec.-PBMCs)	Mixed Chimerism	Grafts
1	8	Control-No treatment	(MCIP)	+
2	8	Control-zipFv (anti CD45RO/	PBMCs + HSCT + (MCIP)	+
		CD127)
3	8	Control-SUPRA CAR T	PBMCs + HSCT + (MCIP)	+
4	8	SUPRA CAR T + zipFv (anti	PBMCs + HSCT + (MCIP)	+
		CD45RO/CD127)
5	8	SUPRA CAR T + zipFv (anti	No PBMCs + HSCT,	+
		CD45RO/CD127)	No MCIP
6	8	SUPRA CAR T + zipFv (anti	No MCIP	+
		CD45RO/CD127)
7	8	SUPRA CAR T + zipFv (anti	⅓ of regular dose MCIP	+
		CD45RO/CD127)

Off-target toxicity occurs when T cells are not able to distinguish normal cells from target cells. In this case CD45RO is the marker that is mainly expressed on the T cells but CD127 is a more common surface marker and can be found on B and dendritic cells. Even though the “Combinatorial AND Logic” CAR system that is described herein has shown to be highly specific, there is a chance for off-target effects in this system. As an alternative approach, all of the CD45RO+ population can be targeted, which consists of not only central memory T cells but also targets effector T cells as well. This will cause more T cells to be targeted nonspecifically but simultaneously might decrease the chance of immune reaction against the graft.

Some of the CART effector cells will express memory phenotype such as CD45RO/CD127 after remaining activated for a period of time. This means that CAR T cells might attack each other after changing phenotype. Long activation of CAR T cells within the patient's body has always been a matter of concern. This hypothetical situation can be considered as a safety step as CART cell will clear out themselves after completing their task which is destroying memory T cells. On the other hand, if turns out to be true, this self-eliminatory phenomenon might increase the number of CAR T cells that are needed to effectively treat the patients.

Benchmark for success: Blood drawn from the vain tail (40 ul) is used every other week for the assessment of peripheral blood chimerism. Monoclonal anti-donor HLA antibody is used for monitoring of chimerism in the various leukocyte lineages, including CD3+/CD8+, CD3+/CD4+, CD3+/CD8−/CD4− (naive T cells), T-Regs, naive and mature B cells (CD19+/CD78- and CD19−/CD78+). Animals are considered chimeric when at least 5% of WBCs in all of these lineages are donor derived. Additionally, the systemic immune status of humanized mice is assessed pre-HSC transplantation and at week 4 and week 8 post-transplant by carboxyfluorescein diacetate succinimidyl ester (CFSE) mixed lymphocyte reaction (MLR) proliferation assays. MLR results are compared before PMBC-HSC transplantation and after study end point.

Determine the Efficacy of pMHC-CAR T-Effector Cells on Specific Depletion of Recipient Alloreactive T Cells and Establishment of Mixed Chimerism.

Described herein is a flexible CAL and/or CAR design that can target any alloantigen specific T cells. Due to the diversity of alloantigen-specific T cells in the context of transplantation, the CAL and/or CAR system that has the flexibility to locate and attack different alloreactive T cells simultaneously. CAL and/or CAR T-eff. cells with (donor) pMHC can target alloreactive T-cells.

The Effect of CAL and/or CAR T Cell Therapy on Targeting a Double Antigen MHC System (OTi and OTii)

In short, pMHC multimer attached to adaptor molecule (e.g., the CAL)+CAR T effector cell is used in the single and double specific antigen-MHC systems. The antigen-specific CAL and/or CAR system can be tested in vitro and in the OTi & ii transgenic mouse model. This model is genetically engineered such that all T cells express receptors that are specific for recognizing chicken ovalbumin peptides (257-264 (OTi) and 329-337 (OTii) in the context of H2Kb (OTi) and I-A b (OTii)). The T cells of these mice models are engineered to express TCRs that specifically recognize pMHC class I/II+Ovalbumin peptides. More importantly, this highly controlled model can be leveraged establish correlation between T cell activities and CAL and/or CAR T cell experimental parameters, such coculture conditions, pMHC affinity, pMHC multimer concentration.

Design, build, and test anti-FITC CAR/pMHC (e.g., the CAL) system in vitro against antigen specific T cells.

As mentioned above, older generations of CAR T cells needed to be genetically redesigned to be able to recognize new or multiple targets. To overcome this challenge, mouse split universal CD8 CAR is generated as described herein. For generating mouse FITC-CAR similar steps that were described previously for human anti-FITC CAR can be utilized 20.To evaluate the efficacy of C67BL/6 mouse anti-FITC CAR T cells efficacy, the experiments that are conducted and mentioned in the section above with human CART cells (FIG. 1-5) are repeated with mouse CART cells.

TABLE 3
outline of experiments

Group	N	Recipient (B6-wild type + OTi/ii)
1	6	Control − No CAR/pMHC
2	6	Control + FITC pMHC multimer
3	6	Control + CAR T (no multimer)
4	6	CAR T + FITC pMHC multimer)

Evaluate the efficacy of CAR T-eff.+FITC-pMHC tetramers (e.g. a CAL) for targeting alloreactive T-cells in the OTi and OTii transgenic mice.

Ovalbumin-pMHC-CAR T-cells effective in recognizing and destroying OTi and OTii T cells a mixture of the OTi and OTii T-cells are transferred to the wild type mice (B6) recipients. The read out of this experiment is the cell count of OTi & ii before and after CAL and/or CAR T cell therapy. The results show the effect of the CAR T-eff cells with the Ovalbumin peptide+MHC tetramer (e.g., the CAL) is successful in targeting the specific clone of OTi & ii cells.

Establish the potency of multiple peptide/multiple MHC (e.g., CAL)−CAR T cell therapy for targeting alloreactive T cell repertoire on mixed chimerism and skin graft survival.

The most significant barrier against graft take is full mismatch across MI-ICs of donors and recipients³². This challenge is addressed herein by using MHC (e.g., CAL)+CART cells to target alloreactive T cells. These MHC multimers are commercially available and can be readily conjugated with FITC labeling to be recognized by the anti FITC CAR T cells. Alternatively, there are many other available options for split universal CAL and/or CAR technology including leucine zippers which provide the option of tuning the affinity of the binding between adaptor (e.g., CAL) and CAR. On the other hand, Minor Histocompatibility Antigens (MiHA) are short immunogenic peptides originating from digested intra/extracellular proteins presented by Major Histocompatibility Complex (human leukocyte antigen). Disparities in minor histocompatibility antigens between individuals who are even MHC matched can induce an immune response after transplantation. These differences have shown to impede tolerance induction through mixed chimerism³³.

Generating a donor-recipient disparities peptide library for evaluating the efficacy of targeting alloreactive T cells in a humanized mouse model.

The role of the default peptides on these exchangeable-peptide MHC systems can be switched with the desired peptides (from the donor) using UV light^34,35. The large collection of peptides associated with human leukocyte antigens (HLA) is referred to as the human immunopeptidome. The desired peptides for this study will are generated by the previously established protocols for immune-peptidome library generation^36,37,38.

Generating tissue-specific immune peptide library from the donor and recipient.

Utilizing previously established protocols we can extract and identify the immunopeptidome of the donor and the recipient. We then identify the peptide sequences that are distinctively different between the donor and the recipient using software tools (below). These disparities in the donors' and the recipients' immune-peptide sequences have the potential to mediate immune rejection. The sequences of the peptides from the donor-recipient disparity peptide library are generated in practical scale by commercially available services³⁹ and loaded on peptide-exchangeable MHC multimers (e.g., a CAL) (QuickSwitch MBL international or Biolegend FlexTetramer) to ultimately be bound to CAR T cells and used for targeting alloreactive T cells in the aforementioned in vitro and in vivo models (FIG. 8, 9). In brief, donor and recipient MHCs associated peptides are isolated independently by immunoaffinity purification using the anti-HLA monoclonal antibodies³⁶. Eluted peptides are identified by different LC-MS/MS systems in DIA (Data Independent A) mode. Mass Spectrometry output files are converted, searched, and statistically validated using software tools (NETMHC, SysteMHC). The identified peptides are then clustered (by GibbsCluster v.1) and annotated by length and predicted MHC binding affinity (NetMHC v.4). The final list of high-confidence donor/recipient MHC-associated peptides is compared between the recipient and the donor and the disparate sequences used for building high-quality donor-recipient disparity peptide library, which is employed as a source for generating peptides to be combined with MBL QuickSwitch (or Bio-legend FlexT) MHC tetramer system.

TABLE 4
outline of experiments

		Recipient (NSG-Hu-		Skin
Group	N	VCA-Rec.-PBMCs)	Mixed Chimerism	Grafts
1	8	Control-No treatment	(MCIP)	+
2	8	SUPRA CAR T + FITC/	PBMCs + HSCT + (MCIP)	+
		(peptide MHC multimer
3	8	Control-SUPRA CAR T	PBMCs + HSCT + (MCIP)	+
4	8	SUPRA CAR T + FITC/	PBMCs + HSCT + (MCIP)	+
		(peptide MHC multimer)
5	8	SUPRA CAR T + FITC/	No PBMCs + HSCT, No	+
		(peptide MHC multimer)	MCIP
6	8	SUPRA CAR T + FITC/	No MCIP	+
		(peptide MHC multimer)
7	8	SUPRA CAR T + FITC/	⅓ of regular dose MCIP	+
		(peptide MHC multimer)

Evaluating the efficacy of targeting alloreactive T cells in the multiple antigen/MHC setting on mixed chimerism establishment in a skin graft humanized mouse model.

In the actual context of allotransplantation, both MHC mismatch and MiHA will be a potential source of immune response and rejection. This experiment addresses the question of whether combing the donor-recipient disparate peptide sequences with exchangeable-peptide MHC system (e.g., a CAL) can be used in combination with split CAR T cells to target alloreactive T cells.

Prior to initiating this experiment, human fetal HSC+skin graft from the same donor is obtained from Advanced Bioscience Resources (CA, US). HLA typing of the donor cells is done and used for generating donor (FITC+exchangeable-peptide) MHC multimer. Peptides that are used with these multimer are generated based on the results of the work described above herein. Briefly, the exchangeable-peptide MHC (e.g., the CAL) is exposed to UV for the dissociation of their default UV-labile peptide. The disparity peptide library provides the desired peptides to replace the default peptide of this system. Using the same double humanized PBMC/HSCT mouse model the efficacy of this donor/recipient disparity peptide library+MHC is evaluated for specific targeting of alloreactive memory T cells and skin allograft survival. As shown in FIGS. 8A-8B, Hu-PBMC treated with exchanged peptide MHC (e.g., CAL)+CAR T. Subsequently, HSCT with fetal HSC is performed and 6 weeks later skin from the same HSC donor is grafted on the back of the mice. Presence of mixed chimerism and graft survival is evaluated using the criteria that was mentioned earlier herein.

The immunopeptidome library is novel and has not previously been adapted for the context of transplantation. As an alternative to immunopeptidome mapping and disparity library generation, we can apply whole-peptidome (donor-recipient) disparity library, a well-known method.

Specificity and killing efficiency of CAL and/or CAR T cells, and chimerism levels can be studied in vitro and in vivo. Peripheral mechanisms of rejection or regulation can be assessed with serial mixed lymphocyte reaction (MLR) assays, at experimental endpoint. For our in vivo studies, our standard clinical definition of tolerance in rodents, which will serve as a benchmark, is rejection and immunosuppression-free survival for 100 days post-HSCT will serve as a benchmark. Skin grafts are assessed as mentioned earlier, based on the color, texture and size of shrinkage.

Impact: Described herein is a novel adaptive immune cell therapy strategy, making use of the CAL and/or CAR T-cell technology, for VCA tolerance induction. Musculoskeletal extremity injuries, including traumatic amputations are frequently seen in the pediatric burn and trauma population. These patients often struggle with severe functional and psychosocial challenges and decreased quality and quantity of life. In turn, the bottleneck to wide application of VCA, with its obvious benefits, would be lifted and effect a paradigm shift in the current management of devastating pediatric burns, which often result in severe disfigurement and amputations. Additionally, successful results enable VCA to be performed for trauma or congenital defects.

REFERENCES

• 1. Ziegler-Graham, K., MacKenzie, E. J., Ephraim, P. L., Travison, T. G. & Brookmeyer, R. Estimating the Prevalence of Limb Loss in the United States: 2005 to 2050. Arch. Phys. Med. Rehabil. 89, 422-429 (2008).
• 2. Hostetler, S. G., Schwartz, L., Shields, B. J., Xiang, H. & Smith, G. A. Characteristics of pediatric traumatic amputations treated in hospital emergency departments: United States, 1990-2002. Pediatrics 116, (2005).
• 3. Brandacher, G., Lee, W. P. A. & Schneeberger, S. Minimizing immunosuppression in hand transplantation. Expert Review of Clinical Immunology vol. 8 673-684 (2012).
• 4. Sachs, D. H., Kawai, T. & Sykes, M. Induction of tolerance through mixed chimerism. Cold Spring Harb. Perspect. Med. 4, 1-19 (2014).
• 5. Spitzer, T. R. et al. Combined histocompatibility leukocyte antigen-matched donor bone marrow and renal transplantation for multiple myeloma with end stage renal disease: The induction of allograft tolerance through mixed lymphohematopoietic chimerism. Transplantation 68, 480-484 (1999).
• 6. Ahmad, S. & Bromberg, J. S. Current status of the microbiome in renal transplantation. Current Opinion in Nephrology and Hypertension vol. 25 570-576 (2016).
• 7. Lellouch, A. G. et al. Abstract 68. Plast. Reconstr. Surg.—Glob. Open 7, 48-49 (2019).
• 8. Leonard, D. A. et al. Vascularized composite allograft tolerance across MHC barriers in a large animal model. Am. J. Transplant. 14, 343-355 (2014).
• 9. Lakkis, F. G. & Sayegh, M. H. Memory T cells: A hurdle to immunologic tolerance. Journal of the American Society of Nephrology vol. 14 2402-2410 (2003).
• 10. Mendenhall, S. D. et al. Prevalence and distribution of potential vascularized composite allotransplant donors, implications for optimizing the donor-recipient match. Plast. Reconstr. Surg.—Glob. Open 6, (2018).
• 11. Yigiter, K. et al. Demography and function of children with limb loss. Prosthet. Orthot. Int. 29, 131-138 (2005).
• 12. Ozalp, B. & Calavul, A. Amputations in burn patients with a special emphasis on pediatric patients. Erciyes Med. J. 39, 7-11 (2017).
• 13. Jang, K. U., Joo, S. Y., Jo, J. H. & Hoon Seo, C. Burn and Amputations: A Retrospective Analysis 379 Amputation out of 19,958 Burns in 10-year. Int. J. Phys. Med. Rehabil. 06, (2018).
• 14. Benichou, G., Gonzalez, B., Marino, J., Ayasoufi, K. & Valujskikh, A. Role of memory T cells in allograft rejection and tolerance. Front. Immunol. 8, (2017).
• 15. Levesque, V. et al. B-cell-dependent memory T cells impede nonmyeloablative mixed chimerism induction in presensitized mice. Am. J. Transplant. (2011) doi:10.1111/j.1600-6143.2011.03683.x.
• 16. Valujskikh, A. The challenge of inhibiting alloreactive T-cell memory. American Journal of Transplantation vol. 6 647-651 (2006).
• 17. Nadazdin, O. et al. Host alloreactive memory T cells influence tolerance to kidney allografts in nonhuman primates. Sci. Transl. Med. 3, (2011).
• 18. Lo, D. J. et al. Selective targeting of human alloresponsive CD8+ effector memory T cells based on CD2 expression. Am. J. Transplant. 11, 22-33 (2011).
• 19. Ng, Z. Y., Read, C., Kurtz, J. M. & Cetrulo, C. L. Memory T Cells in Vascularized Composite Allotransplantation. Vasc. Compos. Allotransplantation 2, 75-79 (2015).
• 20. Tonsho, M. et al. Tolerance of Lung Allografts Achieved in Nonhuman Primates via Mixed Hematopoietic Chimerism. Am. J. Transplant. (2015) doi:10.1111/ajt.13274.
• 21. Hofmann, K., Clauder, A. K. & Manz, R. A. Targeting B cells and plasma cells in autoimmune diseases. Frontiers in Immunology (2018) doi:10.3389/fimmu.2018.00835.
• 22. Ehlers, M. R. & Rigby, M. R. Targeting Memory T Cells in Type 1 Diabetes. Current Diabetes Reports (2015) doi:10.1007/s11892-015-0659-5.
• 23. Rezvani, K., Rouce, R., Liu, E. & Shpall, E. Engineering Natural Killer Cells for Cancer Immunotherapy. Molecular Therapy (2017) doi: 10.1016/j.ymthe.2017.06.012.
• 24. Fesnak, A. D., June, C. H. & Levine, B. L. Engineered T cells: The promise and challenges of cancer immunotherapy. Nature Reviews Cancer vol. 16 566-581 (2016).
• 25. June, C. H., O'Connor, R. S., Kawalekar, 0. U., Ghassemi, S. & Milone, M. C. CART cell immunotherapy for human cancer. Science vol. 359 1361-1365 (2018).
• 26. Cho, J. H., Collins, J. J. & Wong, W. W. Universal Chimeric Antigen Receptors for Multiplexed and Logical Control of T Cell Responses. Cell 173, 1426-1438.ell (2018).
• 27. Yang, J. et al. Allograft rejection mediated by memory T cells is resistant to regulation. Proc. Natl. Acad. Sci. U.S.A. 104, 19954-19959 (2007).
• 28. Levesque, V. et al. B-cell-dependent memory T cells impede nonmyeloablative mixed chimerism induction in presensitized mice. Am. J. Transplant. 11, 2322-2331 (2011).
• 29. Farber, D. L., Yudanin, N. A. & Restifo, N. P. Human memory T cells: Generation, compartmentalization and homeostasis. Nature Reviews Immunology vol. 14 24-35 (2014).
• 30. Thome, J. J. C. et al. Spatial map of human t cell compartmentalization and maintenance over decades of life. Cell 159, 814-828 (2014).
• 31. Antibodies directed against cd127.
• 32. Yong, K. S. M., Her, Z. & Chen, Q. Humanized Mice as Unique Tools for Human-Specific Studies. Archivum Immunologiae et Therapiae Experimentalis (2018) doi:10.1007/s00005-018-0506-x.
• 33. Xia, J. et al. Modeling Human Leukemia Immunotherapy in Humanized Mice. EBioMedicine 10, 101-108 (2016).
• 34. Pakyari, M. et al. A new method for skin grafting in murine model. Wound Repair Regen. 24, 695-704 (2016).
• 35. Murray, A. G. et al. Human T-cell-mediated destruction of allogeneic dermal microvessels in a severe combined immunodeficient mouse. Proc. Natl. Acad. Sci. U.S.A. (1994) doi:10.1073/pnas.91.19.9146.
• 36. Morris, H. et al. Tracking donor-reactive T cells: Evidence for clonal deletion in tolerant kidney transplant patients. Sci. Transl. Med. (2015) doi:10.1126/scitranslmed.3010760.
• 37. Birnbaum, M. E. et al. Deconstructing the peptide-MHC specificity oft cell recognition. Cell (2014) doi:10.1016/j.cell.2014.03.047.
• 38. Gee, M. H. et al. Antigen Identification for Orphan T Cell Receptors Expressed on Tumor-Infiltrating Lymphocytes. Cell (2018) doi:10.1016/j.cell.2017.11.043.
• 39. Ledsgaard, L., Kilstrup, M., Karatt-Vellatt, A., McCafferty, J. & Laustsen, A. H. Basics of antibody phage display technology. Toxins (2018) doi:10.3390/toxins10060236.
• 40. Luimstra, J. J. et al. A flexible MHC class I multimer loading system for large-scale detection of antigen-specific T cells. J. Exp. Med. 215, 1493-1504 (2018).
• 41. Cherf, G. M. & Cochran, J. R. Applications of yeast surface display for protein engineering. Methods Mol. Biol. (2015) doi:10.1007/978-1-4939-2748-7_8.

Example 9

Jurkat+pMHC—OTi

Described herein is the investigation of whether killer CART cells activation by binding to the adaptor (pMHC, e.g, a CAL))+ Target. The experimental design is depicted in FIG. 11.

FITC-conjugated tetramer mediated activation was verified (FIG. 12) and followed in a time course (FIG. 13). Anti-FITC Jurkat vs OT-I experiments were conducted using 100 k cells anti-FITC CAR Jurkat, 100 k cells splenocytes from OT-I mouse, and 5 ug/mL to 78 ng/mL FITC-conjugated tetramer (positive, H2Kb-SIINFEKL (SEQ ID NO: 2750); negative, I-Ab-AAHAEINEA (SEQ ID NO: 2751)). Total Jurkat cell counts at 24 hours (FIG. 14) and tetramer staining levels (FIG. 15) were also determined. The levels of tetramer and CD69 were also determined (FIG. 16).

This experiment demonstrated that Jurkat cells get activated in a dose dependent fashion up to 1 ug/ml of tetramer and OTi binding and staining is dose dependent. No cytotoxicity effects were seen on the killer cells and no prominent activation was seen on the target cells.

Primary T Cells—OTi

Experimental design is shown in FIG. 17. Human CD8 pMHC-CAR T killing is highly specific and MHC-CAR T cell does not kill bystander CD4 T cells (FIG. 18). No cytotoxicity effect was seen on human CD8 pMHC-CAR T (FIG. 19). Cytotoxicity against CAR T cells was long-lived (FIG. 20) and levels of CD69 on OTi CD8 T cells was followed (FIG. 21).

This experiment demonstrated activation of target cells with no significant cytotoxicity on the killer cells. It is possible that Killer Cells activation might be suboptimal under these conditions and use of primary T cells could include fresh, syngeneic cells for improved activation.

Example 10

1E6 T cell clone is a well-established CD8+specific for the major β-cell autoantigen, preproinsulin (PPI). The 1E6 clone mediates β-cell-specific killing via recognition of a highly distinctive HLA A*0201-presented signal peptide epitope (PPI15-24) that exhibits glucose-dependent presentation on the surface of human β-cells (9). The inventors generated and tested a split pMHC-CAR that targets 1E6 T-cells through A*0201+peptides. Effective killing of 1E6 T-cells was observed (FIG. 29)

Example 11

CALs comprising ovalbumin MiHA (against OTi or OTii) were constructed (FIG. 26). pMHC tetramer+FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion, while OTii specific tetramer does not (FIG. 27). Binding of Jurkat+pMHC to OTi cells does not change Jurkat live count (FIGS. 28B and 30). However, human CD8 CAR T were cytotoxic with high specificity (FIGS. 31 and 32). Cytotoxicity was not seen with ctrl tetramer. No cytotoxicity was seen on killer CAR T cells or bystander CD4 T Cells after co-culturing with pMHC and splenocytes.