Cal-t constructs and uses thereof

Abstract

To provide compositions comprising components of multi-component CALs or CARs, and to provide methods for treating autoimmune diseases and the like.SOLUTION: A composition is provided comprising a TCR recognition domain and one or both of (a) an intracellular signaling domain and (b) a first-type protein interaction domain. Further provided is a method for treating or preventing autoimmune disease, transplant rejection, or graft-versus-host disease.SELECTED DRAWING: Figure 10

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62 / 916,924, filed October 18, 2019, the contents of which are incorporated herein by reference in their entirety.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format, which is incorporated by reference in its entirety. This ASCII copy, created on October 16, 2020, is named 701586-096030WOPT_SL.txt and is 540,018 bytes in size.

[0003] Technical Field The technology described herein relates to chimeric antigen ligand (CAL) technology, for example, for treating autoimmune and / or T-cell mediated conditions. [Background technology]

[0004] background The main cause of autoimmunity and graft rejection (e.g., after organ transplantation) is autoreactive and alloreactive T cells that attack the patient's or donor's organs, tissues, and / or cells. Current treatments for preventing autoimmunity and graft rejection require severe immunosuppression, which can lead to unnecessary side effects such as extremely high susceptibility to infection, malignant and benign neoplasms, and multiorgan and multiorgan system failure. To prevent autoimmunity and graft rejection without inducing unnecessary side effects, it would be beneficial to delete autoreactive and alloreactive T cells in patients undergoing organ transplantation without requiring the administration of additional immunosuppressive drugs or a reduction in the dose of the concurrently delivered immunosuppressive regimen. However, this type of treatment has not yet been successfully demonstrated.

[0005] T cells recognize their respective target cells using receptors on their cell surface called T cell receptors (TCRs). TCRs have a recognition portion and a signaling portion. When the recognition portion binds to a natural complex formed in the body in the presence of diseased cells, the signaling portion is activated, and the T cell engages in killing activity or recruits other immune cells to destroy the diseased cells. This natural complex is also known as the peptide-major histocompatibility complex (pMHC complex). CAR-T cell therapy is known and aims to help T cells recognize autoreactive and alloreactive T cells. This is achieved by genetically modifying T cells so that they express chimeric antigen receptors (CARs). CARs are modified TCRs in which the natural recognition portion has been removed and replaced with a synthetic recognition portion designed to more effectively recognize autoreactive and alloreactive T cells by highly specifically detecting the presence of molecules unique to autoreactive and alloreactive T cells. These CAR-T cells are then administered to patients. In the patient's body, the synthetic CAR molecules bind to autoreactive and alloreactive T cells, activating them during this binding process, resulting in the patient's own immune system attacking the diseased T cells. Summary of the Invention

[0006] overview Described herein are methods and compositions related to chimeric antigen ligands (CALs). The CALs described herein comprise a TCR recognition domain and a biomolecular interaction domain. The TCR recognition enables the CAL to antigen-specifically bind to autoreactive and / or alloreactive T cells. The biomolecular interaction domain enables immune killer cells (e.g., NK cells, T cells, or dendritic cells) to bind to the CAL, thereby promoting the killing of autoreactive and / or alloreactive T cells by the immune killer cells.

[0007] The TCR recognition domain of a CAL specifically binds 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, for example, monomeric, oligomeric, and multimeric peptide-MHC complexes. The TCR recognition domain can comprise a natural sequence or a synthetic sequence. Specific examples of peptide-MHC complexes containing autoreactive or alloreactive antigens are provided elsewhere herein.

[0008] The biomolecule interaction domain of the CAL allows the CAL to specifically bind to a second biomolecule (e.g., a receptor on an immune killer cell). In some embodiments, the biomolecule interaction domain of the CAL is recognized by an endogenous receptor on an immune killer cell. In some embodiments, the biomolecule interaction domain of the CAL is recognized by a modified receptor on an immune killer cell. Exemplary, but non-limiting, biomolecular interaction domains include FITC (which can be recognized by FITC CAR-T cell lines), leucine zipper domains, zinc finger domains, PSD95-Dlg1-zo-1 (PDZ) domains, streptavidin and streptavidin-binding protein (SBP) domains, gibberellin insensitive (GIA) and / or gibberellin insensitive dwarf1 (GID1), PYL and ABI domains, chemically derived interaction domain pairs described elsewhere herein, Snap tags, Halo tags, the C-terminal peptides of T14-3-3-cΔC and / or PMA2 (CT52), (R)-phycoerythrin (R-PE / PE) and / or R-PE / PE binding proteins, Fab domains, and / or anti-CD3 domains.

[0009] As mentioned above, CAL described herein can be used in combination with CAR-T system.The exemplary but non-limiting CAR-T system suitable for use with the methods and compositions described herein includes SUPRA CAR and SPLIT CAR.CAR-T system will be discussed in more detail elsewhere in this specification, and is known in the art.

[0010] In one aspect of any embodiment, 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).

[0011] In one aspect of any embodiment, described herein is a composition comprising a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain, and a signaling polypeptide comprising a second type of biomolecular interaction domain and an intracellular signaling domain, wherein the first type and the second type of biomolecular interaction domain specifically bind to each other. In one aspect of any embodiment, described herein is a composition comprising a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain, and a recognition polypeptide comprising a second recognition domain and a third type of biomolecular interaction domain, wherein the first type and the third type of biomolecular interaction domain specifically bind to each other. In one aspect of any embodiment, described herein is a composition comprising a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain, a signaling polypeptide comprising a second type of biomolecular interaction domain and an intracellular signaling domain, and a recognition polypeptide comprising the second recognition domain and a third type of biomolecular interaction domain, wherein the second type and the third type of biomolecular interaction domain compete for binding to the first type of biomolecular interaction domain. In some embodiments of any aspect, the third type biomolecular interaction domain and the first type biomolecular interaction domain have a higher affinity for each other than the second type biomolecular interaction domain and the first type biomolecular interaction domain.

[0012] In one aspect of any embodiment, the present specification describes a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; a signaling polypeptide comprising a second type of biomolecular interaction domain, a fourth type of biomolecular interaction domain, and an intracellular signaling domain; and a recognition polypeptide comprising a second recognition domain and a fifth type of biomolecular interaction domain, wherein the first type of biomolecular interaction domain and the second type of biomolecular interaction domain specifically bind to each other, and the fourth type of biomolecular interaction domain and the fifth type of biomolecular interaction domain specifically bind to each other. In some embodiments of any aspect, the fourth type of biomolecular interaction domain and the fifth type of biomolecular interaction domain have a weaker affinity than the second type of biomolecular interaction domain and the first type of protein interaction domain. In some embodiments of any aspect, the first polypeptide further comprises a sixth type of biomolecular interaction domain, and the recognition polypeptide further comprises a seventh type of biomolecular interaction domain, and they specifically bind to each other.

[0013] In some embodiments of any aspect, the first polypeptide comprises the entire TCR recognition domain. In some embodiments of any aspect, the TCR recognition domain comprises at least two distinct polypeptide sequences, the first polypeptide comprises at least one of the distinct polypeptide sequences of the TCR recognition domain, and the first polypeptide binds to or complexes with a second or additional polypeptide sequence of the TCR recognition domain to form the TCR recognition domain. In some embodiments of any aspect, the TCR recognition domain comprises a non-polypeptide component.

[0014] In some embodiments of any aspect, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain. In some embodiments of any aspect, the second recognition domain is specific for a target that is found on healthy cells and / or non-target cells and not on diseased cells and / or target cells.

[0015] In some embodiments of any aspect, the TCR recognition domain comprises an MHC (major histocompatibility complex), an MHC-peptide complex, a featureless peptide MHC, or an MHC-peptide fusion. In some embodiments of any aspect, the peptide is a human peptide or a non-human peptide. In some embodiments of any aspect, the peptide is a minor histocompatibility antigen (MiHA). In some embodiments of any aspect, the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. In some embodiments of any aspect, the MHC-peptide complex is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. In some embodiments of any aspect, the MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. In some embodiments of any aspect, the MHC is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. In some embodiments of any aspect, the MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. In some embodiments of any aspect, the MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. In some embodiments of any aspect, the MHC is MHC class I or MHC class II.

[0016] In some embodiments of any aspect, the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion. In some embodiments of any aspect, the CD1 is CD1d.

[0017] In some embodiments of any aspect, the biomolecular interaction domain is found in the extracellular portion of the respective polypeptide. a. the biomolecular interaction domain is a leucine zipper or any biomolecular interaction domain binding pair is collectively a leucine zipper pair; b. the biomolecular interaction domain is BZip(RR) and / or AZip(EE), or either biomolecular interaction domain binding pair is collectively BZip(RR) and AZip(EE); c. The biomolecular interaction domain is a PSD95-Dlg1-zo-1 (PDZ) domain; d. the biomolecular interaction domain is streptavidin and / or a streptavidin-binding biomolecule (SBP), or the binding pair of either biomolecular interaction domain is collectively streptavidin and a streptavidin-binding biomolecule (SBP); e. the biomolecular interaction domain is the FKBP binding domain (FRB) of mTOR and / or the FK506 binding biomolecule (FKBP), or either biomolecular interaction domain binding pair is the FKBP binding domain (FRB) of mTOR and the FK506 binding biomolecule (FKBP) as a whole; f. the biomolecular interaction domain is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and / or an FK506-binding biomolecule (FKBP), or either biomolecular interaction domain binding pair is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and an FK506-binding biomolecule (FKBP) as a whole; g. the biomolecular interaction domain is calcineurin A (CNA) and / or FK506-binding biomolecule (FKBP), or the binding pair of either biomolecular interaction domain is calcineurin A (CNA) and FK506-binding biomolecule (FKBP) as a whole; h. the biomolecular interaction domain is gibberellin insensitive (GIA) and / or gibberellin insensitive dwarf1 (GID1), or the binding pair of either biomolecular interaction domain is gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) as a whole; i. the biomolecular interaction domain is a Snap tag and / or a Halo tag, or either biomolecular interaction domain binding pair is collectively a Snap tag and a Halo tag; j. the biomolecular interaction domain is T14-3-3-cΔC and / or the C-terminal peptide of PMA2 (CT52), or the binding pair of either biomolecular interaction domain is T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52) as a whole; k. the biomolecular interaction domain is a PYL and / or an ABI, or either biomolecular interaction domain binding pair is a PYL and an ABI as a whole; l. the biomolecular interaction domain is a nucleotide tag and / or a zinc finger domain, or either biomolecular interaction domain binding pair is collectively a nucleotide tag and a zinc finger domain; m. the biomolecular interaction domain is a nucleotide tag, or any biomolecular interaction domain binding pair is collectively a pair of nucleotide tags; n. the biomolecular interaction domain is fluorescein isothiocyanate (FITC) and / or a FITC-binding biomolecule, or any binding pair of the protein interaction domain is FITC and a FITC-binding protein as a whole; and / or o. The protein interaction domain is (R)-phycoerythrin (R-PE / PE) and / or an R-PE / PE-binding protein, or any binding pair of the protein interaction domain as a whole is (R)-phycoerythrin (R-PE / PE) and an R-PE / PE-binding protein. In some embodiments of any aspect, the nucleotide tag is a DNA tag or a dsDNA tag.

[0018] In some embodiments of any aspect, 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.

[0019] In one aspect of any embodiment, described herein is a cell comprising and / or expressing a composition described herein. In one aspect of any embodiment, described herein is a composition comprising a first polypeptide of any preceding claim and a cell that expresses or comprises a signaling polypeptide of any preceding claim.

[0020] In some embodiments of any aspect, the TCR recognition domain comprises an MHC that is allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any aspect, the TCR recognition domain comprises a synthetic MHC. In some embodiments of any aspect, the TCR recognition domain comprises an MHC and a peptide, and the peptide is allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any aspect, the TCR recognition domain comprises an MHC and a peptide, and the peptide is a synthetic peptide.

[0021] In some embodiments of any aspect, the cells are NK cells, dendritic cells, regulatory T cells, or effector T cells. In some embodiments of any aspect, the cells are modified to express one or more of the polypeptides of the composition. In some embodiments of any aspect, the cells are modified to express the signaling polypeptide of the composition. In some embodiments of any aspect, the cells are further modified to knock out or knock down native MHC I / II. In some embodiments of any aspect, the cells are further modified to knock down native MHC I / II expressed on the cell surface.

[0022] In one aspect of any embodiment, described herein is a composition comprising a TCR recognition domain and an intracellular signaling domain, but not a biomolecular interaction domain (e.g., a first type of biomolecular interaction domain). In one aspect of any embodiment, described herein is a composition comprising a TCR recognition domain and a biomolecular interaction domain (e.g., a first type of biomolecular interaction domain), but not an intracellular signaling domain.

[0023] In one aspect of any embodiment, described herein is a method of treating or preventing an autoimmune disease or condition, a T cell-mediated inflammation or immune response, a malignant T cell condition, a graft rejection, or GvHD in a subject in need thereof, comprising administering to the subject a composition and / or cell of any preceding claim. In some embodiments of any aspect, the TCR recognition domain comprises an MHC allogeneic to the subject.

[0024] In some embodiments of any aspect, the TCR recognition domain comprises an MHC that is autologous to the transplanted cells. In some embodiments of any aspect, the TCR recognition domain comprises an MHC that is xenogeneic to the transplanted cells. In some embodiments of any aspect, the TCR recognition domain comprises an MHC and a peptide, and the peptide is allogeneic to the subject. In some embodiments of any aspect, the TCR recognition domain comprises an MHC and a peptide, and the peptide is autologous to the transplanted cells. In some embodiments of any aspect, the TCR recognition domain comprises an MHC and a peptide, and the peptide is autologous to the transplanted cells. In some embodiments of any aspect, the graft is any human or non-human cell, tissue, or organ. In some embodiments of any aspect, the transplant is an allogeneic hematopoietic stem cell or solid organ transplant.

[0025] In some embodiments of any aspect, the malignant T-cell condition is T-cell acute lymphoblastic leukemia or T-cell lymphoblastic lymphoma.

[0026] In some embodiments of any aspect, the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma. In some embodiments of any aspect, the autoimmune disease is thyroiditis, type 1 diabetes, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barré syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis, e.g., 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, spondyloarthritis, and juvenile-onset rheumatoid arthritis, chronic progressive arthritis, arthritis deformans, primary chronic polyarthritis, and / or juvenile-onset rheumatoid arthritis. primaria), reactive arthritis, and ankylosing spondylitis), relapsing arthritis, inflammatory hyperproliferative skin diseases, psoriasis, such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and nail psoriasis, atopy including atopic diseases, such as hay fever and Job's syndrome, dermatitis, such as contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, nonspecific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic eye inflammatory diseases, urticaria, e.g. chronic allergic urticaria and chronic idiopathic urticaria, e.g. chronic autoimmune urticaria, myositis, polymyositis / dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic sclerosis), sclerosis, e.g. systemic sclerosis, multiple sclerosis (MS), e.g. spino-optical MS, primary progressive MS (PPMS), and relapsing-remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, sclerosis disseminata, ataxic sclerosissclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (e.g., Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis, e.g., ulcerative colitis, colitis ulcerosa, microscopic colitis, collagenous colitis, polypoid colitis, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), enteritis, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome, e.g., adult or acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, autoimmune blood disorders, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as seen in meningitis, herpes gestationis, pemphigoid of gestationis, scrotal pruritus scroti), autoimmune premature ovarian failure, sudden hearing loss due to autoimmune conditions, 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, non-granulomatous uveitis, phacoantigenic uveitis, posterior uveitis or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome, such as chronic or acute glomerulonephritis, e.g. primary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membrano- or membranous nephropathy, including types I and II. proliferative GN (MPGN), and rapidly progressive GN, proliferative nephritis, autoimmune polyendocrine deficiency, balanitis, e.g., plasma cell localized balanitis, balanoposthitis, erythema annulare centrifugally, erythema dyspigmentosa perstans, erythema multiforme, granuloma annulare, lichen sclerosus and atrophicus, lichen simplex chronicus, lichen spinous, lichen planus, ichthyosis lamellar, epidermolytic keratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, allergic reactions, eczema, e.g., allergic eczema or atopic eczema, asteatotic eczema, dyshidrotic eczema, and vesicular palmoplantar eczema, asthma, e.g., bronchial asthmabronchiale, bronchial asthma, and autoimmune asthma, conditions involving T cell infiltration and chronic inflammatory responses, immune responses to foreign antigens such as fetal ABO blood group during pregnancy, chronic pulmonary inflammatory diseases, autoimmune myocarditis, leukocyte adhesion deficiency, lupus, e.g., lupus nephritis, lupus encephalitis, pediatric lupus, non-renal lupus, extrarenal lupus, discoid lupus and discoid lupus erythematosus, lupus alopecia lupus), systemic lupus erythematosus (SLE), e.g., cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and disseminated lupus erythematosus, juvenile-onset (type I) diabetes, e.g., insulin-dependent diabetes mellitus (IDDM) of childhood, adult-onset diabetes mellitus (type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic aortopathy, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, sarcoidosis, granulomatous diseases, e.g., lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitis, e.g., vasculitis, large vessel vasculitis (polymyalgia rheumatica and necrotizing vasculitides such as giant cell (Takayasu's) arteritis, medium-sized vasculitis (including Kawasaki disease and polyarteritis nodosa / periarteritis nodosa), microscopic polyarteritis, immune-mediated vasculitis (immunovasculitis), CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, systemic necrotizing vasculitis, and ANCA-associated vasculitis, e.g., Churg-Strauss vasculitis or Churg-Strauss syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs-positive anemia, Diamond-Blackfan anemia, hemolytic anemia or immune-mediated hemolytic anemia, e.g., autoimmune hemolytic anemia (AIHA), pernicious anemia (pernicious anemia)perniciosa), Addison's disease, pure red cell anemia or pure red cell aplasia (PRCA), factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases with leukocyte leakage, CNS inflammatory disorders, multiple organ injury syndromes e.g. sepsis, secondary to trauma or hemorrhage, antigen-antibody complex mediated diseases, antiglomerular basement membrane disease, antiphospholipid syndrome, allergic neuritis, Behcet's disease / syndrome, Castleman syndrome, Goodpasture's syndrome, Raynaud's syndrome, Sjögren's syndrome, Stevens-Johnson syndrome, pemphigoid e.g. bullous pemphigoid and cutaneous pemphigoid, pemphigus (pemphigus vulgaris, pemphigus foliaceus, mucous membrane pemphigoid) pemphigoid, and pemphigus erythematosus), autoimmune polyendocrinopathy, Reiter's disease or Reiter's syndrome, immune complex diseases such as immune complex nephritis, antibody-associated nephritis, chronic neuropathies such as polyneuropathy, IgM polyneuropathy or IgM-associated neuropathy, and autoimmune or immune-mediated thrombocytopenia, e.g. idiopathic thrombocytopenic purpura (ITP), including chronic ITP or acute ITP, scleritis, e.g. idiopathic ceratoscleritis, episcleritis, autoimmune diseases of the testes and ovaries, e.g. autoimmune seminiferous leukemia, thyroiditis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine disorders including thyroiditis, e.g. autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis) or subacute thyroiditis, idiopathic hypothyroidism, Graves' disease, polyglandular syndromes, e.g. autoimmune polyglandular syndrome (or polyendocrinopathy syndrome), paraneoplastic syndromes including neurological paraneoplastic syndromes, e.g. Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis, e.g. allergic encephalomyelitis or allergic encephalomyelitisAllergic encephalomyelitis (EAE) and experimental allergic encephalomyelitis (EAE), myasthenia gravis including 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, lymphocytic interstitial pneumonia (LIP), bronchiolitis obliterans (non-transplant) vs. NSIP (bronchiolitis obliterans (non-transplant) vs. NSIP) NSIP), Guillain-Barré syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis, e.g. primary biliary cirrhosis and pulmonary cirrhosis, autoimmune enteropathy syndrome, celiac disease or celiac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, amyotrophic lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear diseases, e.g. autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis, e.g. refractory or relapsing or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome / non-syphilitic interstitial keratitis, Bell's palsy, Sweet's disease / Sweet's syndrome, autoimmune rosaceaautoimmune disease), shingles-associated pain, amyloidosis, non-cancerous lymphocytosis, primary lymphocytosis including monoclonal B-cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathy, paraneoplastic syndromes, channelopathies including CNS channelopathies, autism, inflammatory myopathy, focal or segmental glomerulosclerosis or focal segmental glomerulosclerosis (FSGS), endocrine ophthalmopathy, retino-uveitis, chorioretinitis, autoimmune liver disease, fibromyalgia, polyendocrine deficiency, Schmidt's syndrome, adrenal inflammation, gastric atrophy, presenile dementia Demyelinating diseases such as dementia, 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 antisperm antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent miscarriage, farmer's lung, erythema multiforme, postcardiotomy syndrome, Cushing's syndrome, bird fancier's lung, allergic granulomatous vasculitis, benign lymphocytic vasculitis, Alport's syndrome, alveolitis, e.g., allergic alveolitis and fibrosing alveolitis, interstitial lung disease, blood transfusion reactions, Samter's syndrome syndrome, Kaplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial pulmonary fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum, erythroblastosis fetalis, eosinophilic fasciitis, Charmant's syndrome, Felty's syndrome, cyclitis, e.g., chronic cyclitis, heterochronic cyclitis, iridocyclitis (acute or chronic), or Fuch's cyclitis, Henoch-Schönlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndrome, Evan's syndrome, autoimmune hypogonadism, Sydenham's chorea, poststreptococcal nephritis, thromboangitis obliteransubiterans, thyrotoxicosis, spinal cord fistula, choroiditis, giant cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplanted organ reperfusion, autoimmune retinopathy, aphthous stomatitis, arteriosclerotic disorders, aspermiogenesis, autoimmune hemolysis, Beck's disease, allergic enteritis, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, rheumatic fever (Febris rheumatica), Hammann-Rich disease, sensorineural hearing loss, regional ileitis (ileitis regionalis), leukopenia, transverse myelitis, primary idiopathic myxedema, sympathetic ophthalmia, acute polyradiculitis acuta, pyoderma gangrenosum, acquired splenic atrophy, vitiligo, toxic shock syndrome, conditions with T cell infiltration, leukocyte adhesion deficiency, cytokines and T lymphocyte-mediated acute and delayed hypersensitivity and associated immune responses, diseases with leukocyte leakage, multi-organ injury syndrome, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathy, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine deficiency, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid sinusitis, frontal sinusitis, maxillary sinusitis or sphenoid sinusitis, eosinophil-related disorders such as eosinophilia, eosinophilic pulmonary infiltrates, eosinophilic myalgia syndrome, Löffler's syndrome, chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, granulomas containing eosinophils eosinophil), seronegative spondyloarthritis, polyglandular autoimmune disease, sclerosing cholangitis, scleral, episcleral, Bruton's syndrome, transient infantile hypogammaglobulinemia, Wiskott-Aldrich syndrome, ataxia-telangiectasia syndrome, vascular ectasia, autoimmune disorders associated with connective tissue diseases, rheumatism, allergic hypersensitivity disorders, glomerulonephritis, reperfusion injury, ischemia-reperfusion injury, lymphomatous tracheobronchitis, inflammatory skin diseases, skin diseases with an acute inflammatory component, and autoimmune uveoretinitis (AUR).

[0027] In some embodiments of any aspect, 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 aspect, the biologic, cell therapy, or gene therapy is adeno-associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.

[0028] In some embodiments of any aspect, the disease is type 1 diabetes, and the TCR recognition domain comprises a sequence having 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 aspect, the disease is vitiligo, and the TCR recognition domain comprises a sequence having at least 80% or at least 95% sequence identity to SEQ ID NO:18, SEQ ID NO:19, and one of SEQ ID NOs: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 aspect, the method is a method of treating and / or preventing GvHD, and the TCR recognition domain comprises a sequence having 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 aspect, the disease is type 1 diabetes, and the TCR recognition domain comprises one or more of SEQ ID NOs:8-17, or 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 aspect, the disease is vitiligo, and the TCR recognition domain comprises SEQ ID NO:18, SEQ ID NO:19, and one of SEQ ID NOs: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 aspect, the method is a method of treating and / or preventing GvHD, and the TCR recognition domain includes HLA-A*0101 and at least one of SEQ ID NOs:2034-2037, or includes HLA-B*0702 and SEQ ID NO:2038, or includes HLA-B*0801 and SEQ ID NO:2039. [Brief explanation of the drawings]

[0029] [Figure 1] Figure 1 demonstrates that pMHC tetramer + FITC (adapter) binds to target cells (OTi) in a dose-dependent manner. I-H2Kb: MHC class I tetramer. I-Ab-: control tetramer. The figure discloses SEQ ID NOs: 2752-2753, 2750, and 2754, respectively, in the order shown. [Figure 2] Figure 2 shows the expression of the activation marker CD69 on OTi cells at different time points (24, 48, and 72 hours). Binding of Jurkat+pMHC to OTi cells does not alter Jurkat live counts (right). The figures disclose SEQ ID NOs: 2750, 2750, and 2754, respectively, in the order shown. [Figure 3] Figure 3 depicts the expression of CD69 on Jurkat CAR, e.g., CAL T cells at different time points (24, 48, 72 hours). The figure discloses SEQ ID NOs: 2750 and 2755, respectively, in the order indicated. [Figure 4]Figure 4 shows the cytotoxicity of primary human CD8 CARs, e.g., CAL T cells, with various concentrations of tetramer (left). Minimal activation of target cells (CAR, e.g., CD69 on CAL T cells and target cells) after co-culture with OTi cells (Figure 2, left). I-Ab: control / H2Kb: MHC-I. The figures disclose SEQ ID NOs: 2750, 2754, and 2750, respectively, in the order shown. [Figure 5] Figure 5 demonstrates that the cytotoxicity of human CD8 CARs, e.g., CAL T, was highly specific and not observed with control tetramers (left). After co-culture with pMHC and splenocytes, no cytotoxicity was observed against killer CARs, e.g., CAL T cells or bystander CD4 T cells. The figure discloses SEQ ID NOs: 2754, 2750, 2754, 2750, 2754, and 2750, respectively, in the order shown. [Figure 6] Figure 6A is a schematic diagram of a SUPRA CAR design applied to obtain a universal CAL. In this design, the cell targeting molecule module is separated from the killer cell. Figure 6B shows that a SUPRA CAR can be designed in which the CD3ζ domain is not linked to the costimulatory domain to obtain the universal CAL described herein. [Figure 7A] Figures 7A-7D depict key features of the SUPRA CAR system that can be applied to universal CARs. Figure 7A demonstrates that zipCAR activation, as demonstrated by IFN-γ production, is tunable by adjusting zipFv concentration, zipper affinity, scFv affinity, and zipCAR expression levels in human primary CD4 T cells. [Figure 7B] Figure 7B demonstrates that the SUPRA CAR system applied to universal CAL can perform combination antigen detection to form an AND gate logic in CD4 T cells. [Figure 7C]Figure 7C demonstrates that a xenograft animal tumor model shows tumor eradication by SUPRA CAR T cells (as demonstrated by luciferase photon flux given off by the tumor cells). [Figure 7D] Figure 7D demonstrates that the SUPRA CAR applied to universal CAL can be used to target different cell types, e.g., CD4 T cells and CD8 T cells, to two different antigens. CD69 expression (a T cell activation marker) was quantified by flow cytometry on CD4 T cells and CD8 T cells (adapted from Cho, Collins, and Wong, Cell. 2018). [Figure 8] FIG. 8 depicts the timeline for creating the double Hu-PBMC-HSCT-skin transplant mouse model. [Figure 9] FIG. 9 summarizes the creation of the double hu-PBMC-HSCT-skin transplant mouse model. [Figure 10] Figure 10 depicts key features of pMHC multimer+CAR, e.g., CAL T cell lines. [Figure 11] Figure 11 is a table of the experimental design. The figure discloses SEQ ID NOs: 2750, 2756, and 2754, respectively, in the order they appear. [Figure 12] Figure 12 shows validation of FITC-conjugated tetramer-mediated activation. The figure discloses SEQ ID NOs: 2750 and 2751, respectively, in the order indicated. [Figure 13] Figure 13 depicts the time course of FITC-conjugated tetramer-mediated activation. The figure discloses, in order of appearance, SEQ ID NOs: 2750, 2751, 2750, 2751, 2750, 2751, 2750, and 2751, respectively. [Figure 14] Figure 14 depicts a graph of Jurkat cell counts, which disclose SEQ ID NOs: 2750 and 2754, respectively, in the order they appear. [Figure 15]Figure 15 depicts tetramer staining. The figure discloses, from left to right and top to bottom, SEQ ID NOs: 2750, 2754, 2751, 2756, 2754, 2750, and 2754. [Figure 16] Figure 16 is a heat map of staining levels displayed in order of appearance for SEQ ID NOs: 2751, 2757, 2754, 2750, 2754, 2751, 2754, 2757, 2750, 2754, 2751, 2754, 2757, 2750, and 2754, respectively. [Figure 17] FIG. 17 is a table of the experimental design. [Figure 18] Figures 18-20 are graphs of cytotoxicity levels. Figure 18 discloses, from left to right, SEQ ID NOs: 2754, 2750, 2754, and 2750. [Figure 19] FIG. 19 discloses SEQ ID NOs: 2754 and 2750, respectively, in the order they appear. [Figure 20] FIG. 20 discloses, from left to right, SEQ ID NOs: 2750, 2754, 2750, and 2754. [Figure 21] Figure 21 depicts the levels of CD69 on OTi CD8 T cells. The figure discloses, from left to right, SEQ ID NOs: 2750, 2754, 2750, and 2754. [Figure 22] FIG. 22 is a schematic diagram of two embodiments of the technology described herein. [Figure 23] FIG. 23 is a schematic representation of the FU-CAL embodiment of the technology described herein. [Figure 24] Figure 24 is a schematic diagram of the CAL-BITE embodiment of the technology described herein. The left diagram represents blinatumomab, which is described in detail in Weiner et al. The Molecular Basis of Cancer 2015 683-694. e3. [Figure 25] FIG. 25 is a schematic representation of the CAL technique for disarming autoreactive T cells. [Figure 26]Figures 26A-B are schematic diagrams of T cell designs. The MHC can be mouse or human. MiHA can be ovalbumin (for OTi or OTii) or a disparate antigen between donor and recipient. [Figure 27] FIG. 27 is a graph demonstrating that pMHC tetramer+FITC (adapter) binds to target cells (OTi) in a dose-dependent manner, whereas OTii-specific tetramers do not bind to target cells. [Figure 28] Figure 28A shows the expression of CD69 on OTi cells at different time points (24, 48, 72 hours), and Figure 28B is a graph demonstrating that binding of Jurkat+pMHC to OTi cells does not alter the number of viable Jurkat cells. [Figure 29] Figure 29 is a graph demonstrating that the cytotoxicity of pMHC-CAR against the 1E6 T cell clone exhibits a dose-dependent relationship. [Figure 30] Figure 30 is a graph demonstrating CAR Jurkat T cell activation (as measured by the CD69 surface marker) after exposure to target cells (OTi or OTii TCR-expressing T cells) and corresponding tetramers (OTi or OTii specific) (n=4). [Figure 31] Figure 31 is a graph demonstrating that the cytotoxicity of human CD8 CAR T was highly specific and not seen with the control tetramer. After co-culture with pMHC and splenocytes, no cytotoxicity was observed against killer CAR T cells or bystander CD4 T cells (n=4). [Figure 32] Figure 32 is a graph demonstrating the cytotoxicity of primary CD8 CAR T cells against OTi TCR T cells with different tetramer concentrations. DETAILED DESCRIPTION OF THE INVENTION

[0030] 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., a polypeptide comprising at least one MHC sequence described herein) and at least one biomolecular interaction domain. The TCR recognition domain is selected to bind to a specific T cell population that is desired to be targeted and / or destroyed, e.g., for therapeutic purposes in T cell-mediated diseases. In some embodiments, the population of target T cells is a population of polyclonal pathogenic T cells. The biomolecular interaction domain is selected to bind to a second cell (e.g., an NK cell), thereby colocalizing the target T cell with the second cell and facilitating or enhancing the inhibition and / or destruction of the target T cell. In various embodiments, the CAL is selected to have high affinity or high avidity for the TCR (e.g., a TCR variable domain).

[0031] Here, CALs can be used with endogenous cells, for example, in some embodiments, modified cells are not administered to a subject. In another embodiment, CALs can be used with modified cells, for example, modified NK cells. In such cases, modified cells can include one or more CARs, for example, CARs that include an extracellular domain with a biomolecular interaction domain that specifically binds to the biomolecular interaction domain of CALs.

[0032] Thus, described herein are chimeric antigen receptors (CARs) in which the recognition and signaling portions of the CAR are separate polypeptides. These two separate polypeptides that comprise the complete CAR can interact using protein interaction domains to form the complete CAR. This enables flexible, modular CAR-T therapies with complex logic computation capabilities, providing a more precise and effective approach to immunotherapy.

[0033] An aspect of any embodiment includes a cell or composition comprising a CAL and / or chimeric antigen receptor (CAR) having multiple components, and / or a multi-component CAL and / or multi-component CAR. A multi-component CAL / CAR is also referred to herein as a SMART CAL / CAR or SUPRA.

[0034] As used herein, a conventional "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., 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 harness the antigen-binding properties of monoclonal antibodies to redirect the specificity and responsiveness of T cells and other immune cells (e.g., NK cells) to selected targets in a non-MHC-restricted manner.

[0035] Non-MHC-restricted antigen recognition confers the ability of CAR-expressing T cells to recognize antigens independent of antigen processing, thereby bypassing a major mechanism of tumor escape. Moreover, when expressed in T cells, CARs advantageously do not dimerize with the endogenous T cell receptor (TCR) α and β chains. Most commonly, the extracellular binding domain of a CAR is composed of a single-chain variable fragment (scFv) obtained by fusing the variable heavy and variable light chain regions of a mouse or humanized monoclonal antibody. Alternatively, scFv derived from a Fab (not derived from an antibody, e.g., obtained from a Fab library) may be used; in various embodiments, this scFv is fused to a transmembrane domain and then to an intracellular signaling domain. "First-generation" CARs include those that provide only CD3 zeta signaling upon antigen binding, while "second-generation" CARs include those that provide both costimulation (e.g., CD28 or CD137) and activation (CD3Q) signals. "Third generation" CARs include those that provide multiple costimulatory (e.g., CD28 and CD137) and activating (CO3Q) domains. In various embodiments, the CAR is selected to have high affinity or avidity for the antigen. Further discussion of CARs can be found, for example, 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 in its entirety.

[0036] As used herein, a "multi-component CAL" refers to a CAL comprising at least two distinct polypeptides, neither of which independently possesses both the ligand recognition and signal transduction activation capabilities. As used herein, a "multi-component CAR" refers to a CAR comprising at least two distinct polypeptides, neither of which independently possesses both the ligand recognition and signal transduction activation capabilities. In some embodiments, the at least two distinct polypeptides each comprise a protein interaction domain that enables the distinct polypeptides to interact, e.g., bind. In some embodiments, one of the at least two distinct polypeptides is a transmembrane polypeptide having an intracellular T cell receptor (TCR) signaling domain, and the second of the at least two distinct polypeptides is an extracellular polypeptide having a ligand binding domain. In some embodiments, a multi-component CAL and / or a multi-component CAR can comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, or more distinct polypeptides.

[0037] Various aspects provided herein provide compositions that include multiple components of a multi-component CAL and / or a multi-component CAR.

[0038] One aspect of the embodiments includes 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 of protein interaction domain. One aspect of the embodiments includes a composition, e.g., a single molecule, comprising a TCR recognition domain and a first type of biomolecule (e.g., protein) interaction domain. Further provided herein are multi-component CALs and / or multi-component CARs comprising a TCR recognition domain and one or both of (a) an intracellular signaling domain and (b) a first type of protein interaction domain. In some embodiments, the composition, e.g., a single molecule, comprising a TCR recognition domain and a first type of biomolecule (e.g., protein) interaction domain does not include an antibody, antibody domain, or antibody reagent.

[0039] Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain, and (b) a signaling polypeptide comprising a second type of protein interaction domain and an intracellular signaling domain, wherein the first type and the second type of protein interaction domain specifically bind to each other. Further provided herein are multi-component CALs and / or multi-component CARs comprising a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain, and (b) a signaling polypeptide comprising a second type of protein interaction domain and an intracellular signaling domain, wherein the first type and the second type of protein interaction domain specifically bind to each other.

[0040] Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain, and (b) a recognition polypeptide comprising a second recognition domain and a third type of protein interaction domain, wherein the first type and the third type of protein interaction domain specifically bind. Further provided herein are multi-component CALs and / or multi-component CARs comprising (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain, and (b) a recognition polypeptide comprising a second recognition domain and a third type of protein interaction domain, wherein the first type and the third type of protein interaction domain specifically bind.

[0041] Another aspect of the embodiments is a composition comprising: (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; (b) a signaling polypeptide comprising a second type of protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising the second recognition domain and a third type of protein interaction domain, wherein the second type of protein interaction domain and the third type of protein interaction domain compete for binding to the first type of protein interaction domain. Further provided herein are multi-component CALs and / or multi-component CARs comprising: (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; (b) a signaling polypeptide comprising the second type of protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising the second recognition domain and a third type of protein interaction domain, wherein the second type of protein interaction domain and the third type of protein interaction domain compete for binding to the first type of protein interaction domain.

[0042] In various embodiments, the third type protein interaction domain and the first type protein interaction domain have a higher affinity for each other than the second type protein interaction domain and the first type protein interaction domain. Affinity can be measured by one of skill in the art using standard methods, e.g., by measuring the equilibrium dissociation constant (K d ) can be measured.

[0043] Another aspect of the embodiments is a composition comprising: (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; (b) a signaling polypeptide comprising a second type of protein interaction domain, a fourth type of protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth type of protein interaction domain, wherein the first type of protein interaction domain and the second type of protein interaction domain specifically bind to each other, and the fourth type of protein interaction domain and the fifth type of protein interaction domain specifically bind to each other. Further provided herein are multi-component CALs and / or multi-component CARs comprising: (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; (b) a signaling polypeptide comprising a second type of protein interaction domain, a fourth type of protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth type of protein interaction domain, wherein the first type of protein interaction domain and the second type of protein interaction domain specifically bind to each other, and the fourth type of protein interaction domain and the fifth type of protein interaction domain specifically bind to each other.

[0044] In various embodiments, the fourth type of protein interaction domain and the fifth type of protein interaction domain have a weaker affinity than the second type of protein interaction domain and the first type of protein interaction domain. Affinity can be measured as described above.

[0045] In various embodiments, the first polypeptide further comprises a protein interaction domain of type 6 and the recognition polypeptide further comprises a protein interaction domain of type 7, which specifically bind to each other.

[0046] In some embodiments, the first polypeptide comprises the entire TCR recognition domain. In some embodiments, the TCR recognition domain comprises at least two distinct polypeptide sequences, and the first polypeptide comprises at least one of the distinct polypeptide sequences of the TCR recognition domain, and the first polypeptide binds to or complexes with a second or further polypeptide sequence of the TCR recognition domain to form the TCR recognition domain.

[0047] In some embodiments, the compositions described herein can comprise multiple copies or instances of a TCR recognition domain. For example, the TCR recognition domain can be multimeric or oligomeric. In some embodiments, the compositions described herein can comprise multiple copies or instances of a first polypeptide described herein.

[0048] In various embodiments, the second recognition domain is specific for a target not recognized by the TCR recognition domain. In one embodiment, the second recognition domain is specific for a target found on healthy cells and / or non-target cells and not on diseased cells and / or target cells.

[0049] As used herein, a "TCR recognition domain" refers to a domain or portion of a polypeptide that can target or specifically bind to a TCR (e.g., a TCR expressed on the surface of a T cell). In some embodiments, the TCR recognition domain can be a TCR variable region (TCR-VR) recognition domain. That is, the TCR recognition domain can target or specifically bind 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, e.g., chemically modified, compared to the wild-type protein. 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 sequence or a non-human sequence.

[0050] The TCR recognition domain can comprise an MHC polypeptide, an MHC polypeptide sequence, and / or a portion of an 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, e.g., chemically modified, compared to the wild-type protein. 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.

[0051] In one embodiment, the TCR recognition domain comprises an MHC (major histocompatibility complex), an MHC-peptide complex, or an MHC-peptide fusion. In some embodiments, the TCR recognition domain can comprise a featureless peptide-MHC, an MHC without a peptide, or any other molecule that targets or specifically binds to the variable region of a TCR.

[0052] The MHC, also known as human leukocyte antigens (HLA), is composed of a set of genes encoding cell surface proteins essential for the adaptive immune system (e.g., the adaptive immune system) to recognize foreign molecules in vertebrates, thereby determining tissue compatibility. The MHC gene family is divided into three subgroups: MHC class I, MHC class II, and MHC class III. Class I MHC molecules contain the β2 microglobulin subunit, which can only be recognized by the CD8 coreceptor. Class II MHC molecules contain the β1 and β2 subunits and can be recognized by the CD4 coreceptor. In this way, MHC molecules chaperone which type of lymphocyte will bind a given antigen with high affinity, since different lymphocytes express different T cell receptor (TCR) coreceptors. The components of the MHC are known in the art and can be easily identified by those skilled in the art. The MHC is described in more detail in Janeway CA 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, each of which is incorporated herein by reference in its entirety. A complete MHC class I complex contains one MHC class I heavy chain, one peptide ligand sequence, and β2 microglobulin.In some embodiments, the TCR recognition domain comprises an MHC class I heavy chain, a peptide ligand sequence, and β2 microglobulin. A complete MHC class II complex comprises an MHC class II α chain, an MHC class II β chain, and a peptide ligand sequence. In some embodiments, the TCR recognition domain comprises an MHC class II α chain, an MHC class II β chain, and a peptide ligand sequence. These three components can be assembled together as separate sequences (e.g., by intramolecular conjugation of multiple peptide molecules) or expressed as a fusion protein with an intervening linker sequence (see, e.g., Schmittnaegel et al., 2016, incorporated herein by reference in its entirety).

[0053] The MHC (major histocompatibility complex), MHC-peptide complex, MHC-peptide fusion, or uncharacterized peptide MHC can be selected based on the disease or condition to be treated / prevented. Specific MHC, peptide, and / or antigen associated with the diseases described herein are known in the art, and those skilled in the art can select appropriate MHC, peptide, and / or antigen. 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 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 herein by reference in its entirety. The specific examples of TCR recognition domains provided herein are illustrative and not limiting. In addition to those described herein, those skilled in the art can identify appropriate autoantigen pMHC, allogeneic peptide MHC, and autologous peptide MHC, for example, from the art and / or donor cells. Such identification is within the skill of those skilled in the art.

[0054] As an illustrative and non-limiting example of compositions and methods related 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, SEQ ID NO:9, and one of SEQ ID NOs:10-17. In some embodiments, the TCR recognition domain can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more sequence identity to SEQ ID NO:8, SEQ ID NO:9, and one of SEQ ID NOs:10-17. In some embodiments, the TCR recognition domain can comprise a sequence having at least 95% sequence identity to SEQ ID NO:8, SEQ ID NO:9, and one of SEQ ID NOs:10-17, and retaining wild-type activity of SEQ ID NO:8, SEQ ID NO:9, and one of SEQ ID NOs:10-17.

[0055] SEQ ID NO:8 HLA-A*0201 (MHC class I heavy chain allele, wild type, human) TIFF2025172886000002.tif36159

[0056] SEQ ID NO:9 β-2 microglobulin (wild type, human) TIFF2025172886000003.tif10159

[0057] SEQ ID NO:10 Preproinsulin 15~24 (wild type, human) ALWGPDPAAA

[0058] SEQ ID NO:11 preproinsulin 15~24 Modified peptide ligand #1 (synthetic, Cole et al., 2016) AQWGPDPAAA

[0059] SEQ ID NO:12 preproinsulin 15~24 Modified peptide ligand #2 (synthetic, Cole et al., 2016) RQWGPDPAAV

[0060] SEQ ID NO:13 Preproinsulin 15~24 Engineered peptide ligand #3 (wild-type, Clostridium asparagiforme, Cole et al., 2016) RQFGPDWIVA

[0061] SEQ ID NO:14 preproinsulin 15~24 Modified peptide ligand #4 (synthetic, Cole et al., 2016) YQFGPDFPIA

[0062] SEQ ID NO:15 preproinsulin 15~24 Modified peptide ligand #5 (synthetic, Cole et al., 2016) RQFGPDFPTI

[0063] SEQ ID NO:16 preproinsulin 15~24 Modified peptide ligand #6 (synthetic, Cole et al., 2016) YLGGPDFPTI

[0064] SEQ ID NO:17 Preproinsulin 15~24 Engineered peptide ligand #7 (wild-type, Bacteroides fragilis; Cole et al., 2016) MVWGPDLYV

[0065] In a further embodiment of compositions and methods related to 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, SEQ ID NO: 19, and one of SEQ ID NOs: 20-22. In some embodiments, the TCR recognition domain can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more sequence identity to SEQ ID NO: 18, SEQ ID NO: 19, and one of SEQ ID NOs: 20-22. In some embodiments, the TCR recognition domain can comprise a sequence that has at least 95% sequence identity to SEQ ID NO:18, SEQ ID NO:19, and one of SEQ ID NOs:20-22, and retains the wild-type activity of SEQ ID NO:18, SEQ ID NO:19, and one of SEQ ID NOs:20-22.

[0066] SEQ ID NO: 18 HLA-A*0201 (MHC class I heavy chain allele, wild type, human, same as previous HLA-A2) TIFF2025172886000004.tif36159

[0067] SEQ ID NO:19 β-2 microglobulin (wild type, human, same as previous β-2 microglobulin) TIFF2025172886000005.tif10159

[0068] SEQ ID NO:20 MART-1 26~35 (wild type, human) ELAGIGILTV

[0069] SEQ ID NO:21 tyrosinase 368~376 (wild type, human) YMDGTMSQV

[0070] SEQ ID NO:22 gp100 209~217 (wild type, human) ITDQVPFSV

[0071] As further illustrative and non-limiting examples, the following MHC and antigen pairs listed in Table 5 are known in the art. In some embodiments, a TCR recognition domain can comprise one or more of the MHC / peptide pairs listed below, e.g., the TCR recognition domain can comprise one of the MHC alleles listed and the corresponding peptide listed. In some embodiments, a TCR recognition domain can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more sequence identity to one of the MHC / peptide pairs listed below, e.g., to one of the MHC alleles listed and the corresponding peptide listed. In some embodiments, a TCR recognition domain can comprise a sequence having at least 95% sequence identity to one of the MHC / peptide pairs listed below, e.g., to one of the MHC alleles listed and the corresponding peptide listed, and retaining wild-type activity of that MHC allele and the corresponding peptide.

[0072] (Table 5) Each row of the table below lists an MHC allele (the sequence of which is available in publicly accessible databases, e.g., NCBI) and an antigen sequence. The source of the antigen and / or associated disease is also indicated in the same row. In some examples listed in Table 5, peptides are antigen mimics, and their origin is indicated. TIFF2025172886000006.tif103158TIFF2025172886000007.tif231150TIFF2025172886000008.tif241158TIFF2025172886000009.tif241169TIFF2025172886000010.tif237148TIFF2025172886000011.tif237147TIFF2025172886000012.tif241144TIFF2025172886000013.tif236166TIFF2025172886000014.tif241169TIFF2025172886000015.tif241148TIFF2025172886000016.tif241157TIFF2025172886000017.tif241147TIFF2025172886000018.tif241148TIFF2025172886000019.tif241160TIFF2025172886000020.tif241157TIFF2025172886000021.tif241144TIFF2025172886000022.tif234148TIFF2025172886000023.tif240144TIFF2025172886000024.tif241148TIFF2025172886000025.tif241164TIFF2025172886000026.tif241159TIFF2025172886000027.tif241150TIFF2025172886000028.tif241157TIFF2025172886000029.tif241146TIFF2025172886000030.tif241147TIFF2025172886000031.tif236166TIFF2025172886000032.tif241170TIFF2025172886000033.tif241157TIFF2025172886000034.tif160146

[0073] As further illustrative and non-limiting examples, the following antigens listed in Table 6 are known in the art to be associated with celiac disease: In some embodiments, the TCR recognition domain can comprise one or more of the peptides listed in Table 6. In some embodiments, the TCR recognition domain can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more sequence identity to one of the peptides in Table 6. In some embodiments, the TCR recognition domain can comprise a sequence having at least 95% sequence identity to one of the peptides in Table 6 and retaining the wild-type activity of that peptide in Table 6.

[0074] (Table 6) Sequences, in order of presentation, are SEQ ID NOs: 1000-2012. TIFF2025172886000035.tif242159TIFF2025172886000036.tif236159TIFF2025172886000037.tif240159TIFF2025172886000038.tif236159TIFF2025172886000039.tif241159TIFF2025172886000040.tif236166TIFF2025172886000041.tif236156TIFF2025172886000042.tif236159TIFF2025172886000043.tif236159TIFF2025172886000044.tif236159TIFF2025172886000045.tif236159TIFF2025172886000046.tif236159TIFF2025172886000047.tif236159TIFF2025172886000048.tif236159TIFF2025172886000049.tif236159TIFF2025172886000050.tif241159TIFF2025172886000051.tif236159TIFF2025172886000052.tif236159TIFF2025172886000053.tif241159TIFF2025172886000054.tif236157TIFF2025172886000055.tif241159TIFF2025172886000056.tif236159TIFF2025172886000057.tif241158TIFF2025172886000058.tif241158TIFF2025172886000059.tif236159TIFF2025172886000060.tif241159TIFF2025172886000061.tif236159TIFF2025172886000062.tif236155TIFF2025172886000063.tif241160TIFF2025172886000064.tif236160TIFF2025172886000065.tif236157TIFF2025172886000066.tif236159TIFF2025172886000067.tif236157TIFF2025172886000068.tif236159TIFF2025172886000069.tif236159 TIFF2025172886000070.tif236155TIFF2025172886000071.tif236157TIFF202517 2886000072.tif236157TIFF2025172886000073.tif241159TIFF2025172886000074 .tif236159TIFF2025172886000075.tif240159TIFF2025172886000076.tif89157.

[0075] In one embodiment, the peptide is a human, non-human, or synthetic / modified peptide, for example, the peptide is a minor histocompatibility antigen (MiHA).

[0076] Exemplary MiHAs are known in the art, for example, in Spierings et al. Tissue Antigens 2014 84:347-360, which is incorporated herein by reference. MiHAs are typically utilized in transplantation-related embodiments, where they refer to epitopes that arise due to protein sequence variations in polymorphic proteins.

[0077] Residues of antigenic peptides that associate with the MHC-binding groove can be broadly divided into two types: anchor residues that associate with MHC molecules to confer stability to the MHC-peptide complex (see, e.g., residues P2, P3, P5, P6, P7, and P9 in Bowness et al. 1999 Expert Reviews in Molecular Medicine 16:1-10, incorporated herein by reference in its entirety), and interface residues that are solvent-exposed and can associate with cognate T cell receptors (see, e.g., residues P1, P4, and P8 in Bowness). A featureless peptide is a peptide in which the anchor residues are preserved but the interface residues of the peptide have been mutated to alanine or glycine residues to prevent TCR binding. Therefore, a featureless peptide-MHC is an MHC-peptide complex in which the presented peptide is a featureless peptide. A featureless peptide-MHC is typically used in embodiments related to transplantation tolerance. In patients undergoing MHC-mismatched solid organ or hematopoietic stem cell transplantation, the use of CAL T cells presenting the appropriate donor-mismatched featureless peptide-MHC CAL can allow for the selective depletion of recipient alloreactive T cells that target the mismatched donor HLA.

[0078] The MHC can be a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. In embodiments where more than one MHC unit is present, e.g., where the MHC is an oligomer, the units can be provided consecutively, e.g., in a chain, or arranged in one or more dimensions around a central point or linker, or conjugated / attached to a scaffold in any geometric arrangement, or any combination thereof. 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 above references is incorporated herein by reference in its entirety.

[0079] In some embodiments, the TCR recognition domain can comprise a CD1 domain (e.g., a CD1d domain), for example, a sequence comprising the 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 presents lipid antigens to T cells. The sequences of several CD1d isoforms and the structure of CD1d are known in the art. See, for example, the NCBI database for CD1d (Gene ID 912) and the three isoforms listed in Bagchi et al., 2018 and Oleinika et al., Nature Communications 2018 9:684, each of which is incorporated herein by reference in its entirety. For example, CD1d isoform 1 is SEQ ID NO:5 (NCBI Ref Seq NP_001757.1), CD1d isoform 2 is SEQ ID NO:6 (NCBI Ref Seq NP_001306074.1), and CD1d isoform 3 is SEQ ID NO:7 (NCBI Ref Seq NP_001358690.1). In some embodiments, the CD1d domain comprises a sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more sequence identity to one of SEQ ID NOs:5-7. In some embodiments, the CD1d domain comprises a sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more 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 having at least 95% sequence identity to one of SEQ ID NOs:5-7 and retaining the lipid binding activity of the wild-type reference sequence.In some embodiments, the CD1d domain comprises a sequence that has 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.

[0080] In some embodiments, the CD1 domain further comprises a ligand, e.g., a non-peptide ligand. Exemplary, non-limiting ligands for the CD1 domain include, but are not limited to, the following: Protein (allele) ligand TIFF2025172886000077.tif61128

[0081] In some embodiments, the TCR recognition domain can include, for example, an MHC, pMHC, or CD1 domain and sequences or molecules other than the same. For example, it can further include other polypeptide and / or non-polypeptide components that enable multimerization. Exemplary components that enable multimerization can include biotin (a non-polypeptide) and / or streptavidin polypeptides used to enable tetramerization.

[0082] In various embodiments, the protein interaction domains are found in the extracellular portion of the respective polypeptides.

[0083] As used herein, a "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.

[0084] As used herein, a "signaling polypeptide" refers to a transmembrane polypeptide having an intracellular signaling domain, such as 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.

[0085] As used herein, a "biomolecular interaction domain" refers to a domain that allows two distinct molecules to specifically bind to each other. These molecules can be or include polypeptides. In some embodiments, one or both of these molecules or biomolecular interaction domains can be non-peptides. When a pair of biomolecular interaction domains is provided herein, they allow two or more molecules to specifically bind, for example, one of the biomolecular interaction domains can specifically bind to the second biomolecular interaction domain. In some embodiments, specific binding can occur, for example, when there are two distinct biomolecular interaction domains of a pair. In some embodiments, specific binding can occur when there are three or more distinct biomolecular interaction domains. Note that a protein interaction domain is one type of biomolecular interaction domain, and where one is specified herein, it can always be replaced with the other.

[0086] As used herein, reference to a molecule as a protein or polypeptide includes protein, peptide, or polypeptide sequences, but may also include non-proteinaceous motifs, modifications, or domains. Some exemplary biomolecular interaction domains, along with protein interaction domain pairs, are listed elsewhere herein. In some embodiments, a biomolecular interaction domain comprises, consists of, or consists essentially of a protein or polypeptide. In some embodiments, a biomolecular interaction domain comprises, consists of, or consists essentially of a non-protein molecule. In some embodiments, one member of a biomolecular interaction domain pair can comprise, consist of, or consist essentially of a protein or polypeptide, while the other member of the biomolecular interaction domain pair can comprise, consist of, or consist essentially of a non-protein molecule (e.g., FITC and anti-FITC). Exemplary protein interaction domains are known in the art and can be used in the aspects described herein.

[0087] As used herein, a "protein interaction domain" refers to a domain that allows specific binding between two distinct polypeptides. Some exemplary protein interaction domains, along with pairs of protein interaction domains, are listed elsewhere herein. In some embodiments, the protein interaction domains of the polypeptides of a multi-component CAL and / or a multi-component CAR can specifically bind, for example, one of the protein interaction domains can specifically bind to another protein interaction domain of the multi-component CAL and / or a multi-component CAR. In some embodiments, specific binding can occur when there are two distinct protein interaction domains. In some embodiments, specific binding can occur when there are three or more distinct protein interaction domains. Exemplary protein interaction domains are known in the art and can be used in the aspects described herein.

[0088] In some embodiments of any of the aspects described herein, the protein interaction domain is a leucine zipper domain. Leucine zipper domains are a type of protein-protein interaction domain commonly found in transcription factors, characterized by leucine residues evenly spaced throughout an alpha helix. Leucine zippers can form heterodimers or homodimers. Several leucine zipper domains and their ability to bind to each other are known in the art. These are further discussed, for example, 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 herein by reference in its entirety. In some embodiments, one leucine zipper domain is BZip(RR) and the other leucine zipper domain is AZip(EE). In some embodiments, the sequence of the BZip(RR) leucine zipper domain is TIFF2025172886000078.tif4158. In some embodiments, the sequence of the AZip(EE) leucine zipper domain is The file is TIFF2025172886000079.tif4158. Further exemplary leucine zipper domains are described in Reinke et al. JACS 2010 132:6025-31, which is incorporated herein by reference in its entirety. For example, suitable leucine zipper domains can include SYNZIP1-SYNZIP48, as well as BATF, FOS, ATF4, ATF3, BACH1, JUND, NFE2L3, and HEPTAD. The binding affinities of various combinations of these domains are described, for example, in Figure 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.

[0089] Further exemplary pairs of protein interaction domains include a) PSD95-Dlg1-zo-1 (PDZ) domains, b) streptavidin domains and streptavidin-binding protein (SBP) domains, and c) PYL domains and ABI domains.

[0090] In some embodiments of any of the aspects described herein, the protein interaction domain can be a chemically derivatized protein interaction domain, e.g., a domain that specifically binds only in the presence of a third molecule, such as a small molecule or small molecule drug. Exemplary chemically induced protein interaction domains include the FKBP-binding domain (FRB) of mTOR and FK506-binding protein (FKBP) (the binding of which is activated by tacrolimus, everolimus, or a rapalog), cyclophilin-Fas fusion protein (CyP-Fas) and FK506-binding protein (FKBP) (the binding of which is activated by FKCsA), calcineurin A (CNA) and FK506-binding protein (FKBP) (the binding of which is activated by FK506), gibberellin-insensitive (GIA) and gibberellin-insensitive dwarf1 (GID1) (the binding of which is activated by gibberellin), Snap tag and Halo tag (the binding of which is activated by HaXS), and T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52) (the binding of which is activated by fusicoccin). Further description of chemically derived protein interaction domains can be found in the art, for example, 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.

[0091] In some embodiments of any of the aspects described herein, the protein interaction domain can comprise at least one nucleotide tag and at least one zinc finger domain. Zinc finger domains are characterized by the coordination of zinc ions to stabilize their tertiary structure. The specific folds exhibited by the zinc fingers can vary. In some embodiments, the zinc finger domain can be a nucleotide-binding zinc finger domain. In some embodiments, the 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, the nucleotide tag can be a DNA tag. In some embodiments, the nucleotide tag can be a dsDNA tag containing the entire recognition sequence of the zinc finger domain being used. Exemplary zinc finger domains and their cognate nucleotide tags are described in the art, for example, in Mali et al. Nature Methods 2013 10:403-406, which is incorporated herein by reference in its entirety. In some embodiments, the zinc finger domain can be sZF15 as described in Mali et al. Nature Methods 2013 10:403-406.

[0092] In one embodiment the protein interaction domain is BZip(RR) and / or AZip(EE), or any binding pair of protein interaction domains as a whole is BZip(RR) and AZip(EE).

[0093] In some embodiments of any of the aspects described herein, the protein interaction domain can comprise a substantially complementary nucleotide tag, for example, fully complementary or sufficiently complementary to specifically hybridize. The degree of complementarity required can vary depending on the total length of the tag and the G / C content of the complementary portion. Those skilled in the art can readily determine the appropriate affinity required for a tag of a given size and / or G / C content. In some embodiments, the nucleotide tag can be a DNA tag.

[0094] In some embodiments, the nucleotide tag can be a DNA tag. In some embodiments, the nucleotide tag can be a dsDNA tag.

[0095] In one embodiment, the protein interaction domain is gibberellin insensitive (GIA) and / or gibberellin insensitive dwarf1 (GID1), or any binding pair of the protein interaction domain as a whole is gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1).

[0096] In one embodiment, the protein interaction domain is a Snap tag and / or a Halo tag, or any binding pair of protein interaction domains as a whole is a Snap tag and a Halo tag.

[0097] In one embodiment, the protein interaction domain is T14-3-3-cΔC and / or the C-terminal peptide of PMA2 (CT52), or any binding pair of protein interaction domains as a whole is T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52).

[0098] In one embodiment, the protein interaction domain is PYL and / or ABI, or any binding pair of protein interaction domains as a whole is PYL and ABI.

[0099] In one embodiment, the biomolecular interaction domain is fluorescein isothiocyanate (FITC) and / or a FITC-binding protein, or either biomolecular interaction domain binding pair as a whole is FITC and a FITC-binding protein.

[0100] In one embodiment, the biomolecular interaction domain is (R)-phycoerythrin (R-PE / PE) and / or an R-PE / PE binding protein, or either biomolecular interaction domain binding pair as a whole is (R)-phycoerythrin (R-PE / PE) and / or an R-PE / PE binding protein.

[0101] In some embodiments, a biomolecular domain on a molecule comprising a TCR recognition domain and a biomolecular interaction domain (i.e., a CAL) specifically binds to a native cell surface molecule on an NK cell, a dendritic cell, or a T cell. In such embodiments, a subject can be treated as described herein without administering modified cells. In some embodiments, a biomolecular domain on a molecule comprising a TCR recognition domain and a biomolecular interaction domain (i.e., a CAL) specifically binds to a native cell surface molecule on an NK cell. In some embodiments, a biomolecular domain on a molecule comprising a TCR recognition domain and a biomolecular interaction domain (i.e., a CAL) specifically binds to a native cell surface molecule on a dendritic cell. An exemplary native cell surface molecule for such embodiments includes, but is not limited to, CD3. Other suitable cell surface molecules are CD cell surface proteins. CDs are known in the art, and one of skill in the art can readily select one expressed by a desired cell type. See, for example, the HCDM database at hcdm.org, and the list 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. Thus, exemplary biomolecular interaction domains for such embodiments include, but are not limited to, anti-CD3 antibody reagents or Fab domains. Those skilled in the art will be aware of other cell surface molecules found on, or exclusively found on, the cell surface of related cell types, as well as multiple reagents capable of binding (e.g., specifically binding) to such cell surface molecules.

[0102] In aspects with a single recognition polypeptide and a single signaling polypeptide that can specifically bind without a third polypeptide, the multi-component CALs or multi-component CARs described herein will be activated in the presence of a target ligand, thereby inducing T cell activity in the vicinity of the target ligand. Further described herein are multi-component CALs or multi-component CARs capable of logical computation, such as multi-component CALs or multi-component CARs that serve as AND, OR, or NOT logic gates.

[0103] In some aspects, the present specification describes compositions containing components of multi-component CALs and / or multi-component CARs that enable AND gate logic.In these aspects, the activation of multi-component CALs and / or multi-component CARs occurs only in the presence of two target ligands, and the recognition of a single target ligand is not sufficient for activation.Such multi-component CALs or multi-component CARs can increase specificity and reduce off-target effects.A single ligand that is a good marker for target cells or target tissues may also exist elsewhere in the subject, which will result in off-target effects.However, if the recognition of two separate marker ligands is required, the possibility of off-target activity is reduced.

[0104] In one embodiment, the nucleotide tag is a DNA tag or a dsDNA tag.

[0105] Further embodiments of AND logic gates multi-component CAL and AND logic gates multi-component CAR are also described herein.

[0106] In some embodiments of any of the aspects described herein, the composition includes a component of a multi-component CAL and / or multi-component CAR that is a NOT logic gate. For example, recognition of a second targeting ligand by a second recognition polypeptide can prevent interaction (e.g., specific binding) between a signaling polypeptide and a first recognition polypeptide. Such embodiments can enable suppression of T cell activity in inappropriate and / or off-target tissues. For example, the second targeting ligand can be a marker for a tissue that is particularly sensitive to T cell activity, a known off-target active region, and / or a marker that shares a common marker with the desired target tissue. In some embodiments, in a NOT-gated multi-component CAL and / or a NOT-gated multi-component CAR, the second targeting ligand is not a ligand found in the target tissue and / or target cells, e.g., in or on diseased T cells. In some embodiments, the second targeting ligand of a NOT-gated multi-component CAL and / or a NOT-gated multi-component CAR is found on healthy cells and / or non-target cells, but not on diseased cells and / or target cells. A variety of two-dimensional and three-dimensional configurations for such pairs of nucleotide pairs are known in the art.

[0107] In some embodiments, the target ligand recognized by the second recognition polypeptide is found on healthy cells and / or non-target cells, but not on diseased cells and / or target cells. In some embodiments, the protein interaction domain of the second recognition polypeptide and the protein interaction domain of the first recognition polypeptide have a stronger 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 stronger 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, for example, by binding affinity assays known in the art, and relative binding affinities are known for some combinations of protein interaction domains described herein. See, e.g., Reinke et al. JACS 2010 132:6025-31, incorporated herein by reference in its entirety. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domain can be at least 2-fold stronger than the binding affinity of the first recognition polypeptide protein interaction domain to the signaling polypeptide interaction domain. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domain can be at least 5-fold stronger than the binding affinity of the first recognition polypeptide protein interaction domain to the signaling polypeptide interaction domain. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domain can be at least 10-fold stronger than the binding affinity of the first recognition polypeptide protein interaction domain to the signaling polypeptide interaction domain.

[0108] As used herein, a "target ligand" refers to a molecule in or on a cell that can be bound by a ligand-binding domain. Non-limiting examples of such molecules can include polypeptides, lipids, sugars, 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.

[0109] In some embodiments, e.g., embodiments involving multicomponent CALs and / or multicomponent CARs with a single recognition polypeptide or AND-gated multicomponent CALs and / or AND-gated multicomponent CARs, the targeting ligand (e.g., the first and / or second targeting ligand) can be a ligand expressed in a target tissue. In some embodiments, the targeting ligand can be constitutively expressed in the target tissue and / or target cell. In some embodiments, the targeting ligand can be expressed exclusively in the target tissue and / or target cell. In some embodiments, the targeting ligand can be expressed at a higher level in the target tissue and / or target cell than in other tissues and / or cells. Because recognition of the targeting ligand in embodiments involving multicomponent CALs and / or multicomponent CARs with a single recognition polypeptide or AND-gated multicomponent CALs and / or AND-gated multicomponent CARs can result in T cell activation (e.g., cell-killing activity of cells containing the targeting ligand), the targeting ligand can be selected to target T cell activity in a desired and / or therapeutic manner, e.g., by targeting diseased cells. In some embodiments, the targeting ligand is a ligand found in / on diseased cells and / or target cells. In some embodiments, the target ligand that is specifically bound by the recognition polypeptide that can specifically bind to the signaling polypeptide or by the recognition polypeptide that is a part of the AND gate multicomponent CAL and / or the AND gate multicomponent CAR is a ligand that is found in / on disease cells and / or target cells. In some embodiments, the target ligand that is specifically bound by the recognition polypeptide that can specifically bind to the signaling polypeptide or by the recognition polypeptide that is a part of the AND gate multicomponent CAL and / or the AND gate multicomponent CAR is a ligand that is found on disease cells and / or target cells and not on healthy cells and / or non-target cells. In some embodiments, the disease cells are autoreactive or alloreactive T cells.In some embodiments, the target ligand that is specifically bound by a recognition polypeptide that can specifically bind to a signaling polypeptide, or by a recognition polypeptide that is part of an AND gated multicomponent CAL and / or an AND gated multicomponent CAR, is found on the surface of diseased cells. In some embodiments, the recognition polypeptide that can specifically bind to a signaling polypeptide, or a recognition polypeptide that is part of an AND gated multicomponent CAL and / or an AND gated multicomponent CAR, specifically binds to the target ligand on the surface of diseased cells, for example, compared to binding to normal cells.

[0110] In some embodiments, the compositions and / or cells described herein can further comprise a second multi-component CAL and / or multi-component CAR according to any of the aspects and embodiments described herein for the first multi-component CAL and / or multi-component CAR. As a non-limiting example, the second CAL and / or CAR can be designed to specifically bind to (and, e.g., be activated or inhibited by) a target ligand different from that to which (e.g., be activated or inhibited by) the first multi-component CAL and / or multi-component CAR. This can increase the specificity, reduce off-target effects, and / or reduce the effective dosage for the methods described herein. In some embodiments, the recognition domain of the second multi-component CAL and / or multi-component CAR specifically binds to a target ligand different from that bound by the recognition domain of the first multi-component CAL and / or multi-component CAR. In some embodiments, the antibody reagent of the second multi-component CAL and / or multi-component CAR specifically binds to a target ligand different from that bound by the antibody reagent of the first multi-component CAL and / or multi-component CAR.

[0111] In some embodiments, the second multi-component CAL and / or multi-component CAR can include an inhibitory intracellular signaling domain, e.g., a T cell receptor (TCR) signaling domain, e.g., one that inhibits the activity of the engineered cell (e.g., engineered T cell). Thus, in such embodiments, while the first multi-component CAL and / or multi-component CAR enables activation of T cell activity, the second multi-component can be designed to act in opposition to the first multi-component CAL and / or multi-component CAR, e.g., to inhibit T cell activation. Inhibitory intracellular TCR signaling domains are known in the art, and non-limiting examples include PD1, CTLA4, BTLA, KIR, LAG-3, TIM-3, A2aR, LAIR-1, and TGIT. In one embodiment, a non-active mimetic of an activating TCR can be used.

[0112] In some embodiments, described herein are compositions comprising a cell, a CAL, and a CAR (e.g., the CAR comprises either an inhibitory domain or a costimulatory / activation domain).

[0113] In one embodiment, the TCR recognition domain comprises an 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 an 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 peptide that is non-self with respect to the target cell.

[0114] In some embodiments, the target ligand specifically bound by the recognition polypeptide that can specifically bind to the signaling polypeptide of the second multi-component CAL and / or multi-component CAR that comprises an inhibitory intracellular signaling domain is a ligand found on healthy cells and / or non-target cells. In some embodiments, the target ligand specifically bound by the recognition polypeptide that can specifically bind to the signaling polypeptide of the second multi-component CAL and / or multi-component CAR that comprises an inhibitory intracellular signaling domain is a ligand found on healthy cells and / or non-target cells and not on diseased cells and / or target cells. In some embodiments, the second multi-component CAL and / or multi-component CAR that comprises 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.

[0115] In some embodiments of any aspect, the 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'), an Fv, a disulfide-linked Fv, a scFv, a single-domain antibody, a diabody, a multispecific antibody, a bispecific antibody, an anti-idiotypic antibody, and / or a bispecific antibody.

[0116] 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 above.

[0117] Multicomponent CAR / CAL cells The present specification provides a cell that expresses the composition or multi-component CAR and / or multi-component CAL provided herein.The cell can be any cell, for example, any mammalian cell, for example, human cell.In one embodiment, the cell is a dendritic cell, a regulatory T cell, or an effector T cell.In one embodiment, the cell is a dendritic cell (CAL DC), a T cell (for example, an effector, regulatory, etc.) (CAL-T), a regulatory T cell, an effector T cell, a natural killer cell (CAL NK), or any other myeloid cell.

[0118] In one aspect, described herein are engineered cells that express and / or comprise one or more multi-component CAR / CALs, or compositions comprising the same described herein, e.g., compositions comprising at least one signaling polypeptide and at least one recognition polypeptide. In some embodiments, the cells are natural killer (NK) cells, dendritic cells, regulatory T cells, or effector T cells. Such cells that express and / or comprise 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, complete multi-component CAR / CAL cells express both a signaling polypeptide (e.g., a CAR) and at least one recognition polypeptide (e.g., a CAL or adapter described elsewhere herein) of a multi-component CAL and / or multi-component CAR. In some embodiments, complete multi-component CAL and / or CAR cells comprise nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and / or multi-component CAR. In some embodiments, the signaling polypeptide is present on the membrane of the cell. In some embodiments, one or more recognition polypeptides are present in the extracellular space. For example, the recognition polypeptide may be expressed and secreted by the cell, or the cell may be contacted with a recognition polypeptide provided from another source (e.g., produced synthetically or produced by another cell and optionally purified or processed prior to the contacting step).

[0119] In any of the aspects described herein, e.g., those relating to either complete multicomponent CAL and / or CAR cells or partial multicomponent CAL and / or CAR cells, the recognition and / or signaling polypeptides 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 that is constantly expressed in 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 that can be expressed in response to a given signal, e.g., the addition or reduction of an agent. A non-limiting example of an inducible promoter is a promoter that is regulated in a specific tissue type. The promoter may be regulated by a steroid hormone, a polypeptide hormone (e.g., by using a signal transduction pathway), or a heterologous polypeptide (e.g., the tetracycline-inducible system "Tet-On" and "Tet-Off"; see, e.g., Clontech Inc. (California), Gossen and Bujard, Proc. Natl. Acad. Sci. USA 89:5547, 1992, and Paillard, Human Gene Therapy 9:983, 1989, each of which is incorporated herein by reference in its entirety). In some embodiments, the expression of the polypeptide can be precisely regulated, for example, by using an inducible regulatory sequence that is sensitive to certain physiological regulators, such as circulating glucose levels or hormones (Docherty et al., 1994, FASEB J. 8:20-24).Such inducible expression systems suitable for controlling expression in cells or mammals include, for example, regulation by ecdysone, estrogen, progesterone, tetracycline, chemical inducers of dimerization, and isopropyl-β-D1-thiogalactopyranoside (IPTG). One skilled in the art would be able to select an appropriate regulatory / promoter sequence based on the intended use of the polypeptide.

[0120] In some embodiments, expression of one or more of the recognition or signaling polypeptides can be constitutive. In some embodiments, expression of one or more of the recognition or signaling polypeptides can be transient. Transient expression can be achieved, for example, by using transient and / or inducible expression promoters, or by using transient vectors, such as those that do not integrate into the genome and / or persist in the target cell. As a non-limiting example, derivatives of viruses such as 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 and general recombinant techniques, see Chapters 16 and 17 of Molecular Cloning: A Laboratory Manual (2nd ed., Sambrook, Fritsch, and Maniatis, eds.) (Cold Spring Harbor Laboratory Press: 1989), incorporated herein by reference in its entirety. In some embodiments, the signaling polypeptides of the multi-component CAL and / or multi-component CAR can be constitutively expressed and the recognition polypeptides can be transiently expressed. In some embodiments, the recognition polypeptides of the multi-component CAL and / or multi-component CAR can be constitutively expressed and the signaling polypeptides can be transiently expressed. In some embodiments, the recognition polypeptides of the multi-component CAL can be constitutively expressed and the signaling polypeptides can be exogenously provided. In some embodiments, the recognition polypeptides of the multi-component CAL can be transiently expressed and the signaling polypeptides can be exogenously provided. In some embodiments, the signaling polypeptides of the multi-component CAL and / or multi-component CAR can be constitutively expressed and the recognition polypeptides can be exogenously provided. In some embodiments, the signaling polypeptides of the multi-component CAL and / or multi-component CAR can be transiently expressed and the recognition polypeptides can be exogenously provided.

[0121] The CARs and CALs described herein can be produced according to any method known in the art, such as recombinant expression or peptide synthesis. Exemplary methods include the NIH Tetramer Core Facility's MHC expression protocol (available on the World Wide Web at tetramer.yerkes.emory.edu / support / protocols#1), ProImmune's pentamer protocol (available on the World Wide Web at proimmune.com / protocols-2 / ), Immudex's dextramer protocol (available on the World Wide Web at immudex.com / resources / protocols / ), and the CAR T cell production protocol published in the "Primary Human T cell Isolation and Culture" and "Lentiviral Transduction of Human T cells" subsections of the "Method Details" section of Cho et al., 2018 Cell 173:1426-1438, each of which is incorporated herein by reference in its entirety.

[0122] In one aspect, described herein are methods of killing target cells, comprising contacting the cells 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 disease cell, e.g., an autoreactive or alloreactive T cell. In one aspect, described herein are methods of treating or preventing a disease, e.g., an autoimmune disease or condition, a T cell-mediated inflammation or immune response, a 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 are methods of treating or preventing an autoimmune disease or condition, a T cell-mediated inflammation or immune response, a transplant rejection, or GvHD, comprising administering a complete multi-component CAR cell, CAR, and / or CAL according to any of the embodiments described herein.

[0123] 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, the method comprising administering a complete multi-component CAR / CAL, CAR and / or CAL 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, the method comprising administering a complete multi-component CAR cell, CAR and / or CAL according to any of the embodiments described herein.

[0124] In some embodiments, the complete multi-component CAL and / or CAR cells can be autologous or allogeneic to the subject. In some embodiments, the complete multi-component CAL and / or CAR cells can be derived from and / or the progeny of cells obtained from the subject or a third party and have been modified ex vivo to contain at least one multi-component CAL and / or multi-component CAR (e.g., genetically modified to contain nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of the multi-component CAL and / or multi-component CAR). In some embodiments, the method can further include obtaining cells (e.g., dendritic cells, regulatory T cells, or effector T cells) from the subject, modifying the cells to contain nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of the multi-component CAL and / or multi-component CAR, and then administering the cells to the subject.

[0125] In one embodiment, the modified cells are further modified to lack or reduce the expression of native MHC I / II, for example, as measured on the cell surface.Methods for modifying cells to reduce or eliminate native MHC I / II are known in the art, including, for example, the expression of RNA interference (such as short hairpin RNA, small interfering RNA, double-stranded RNA), the expression of inhibitory oligonucleotides, nuclease-based inhibition (such as CRISPR, TALEN, meganuclease, etc.), and the expression of KDEL motif (SEQ ID NO: 2749) containing binding proteins capable of confining MHC I / II to the endoplasmic reticulum.Those skilled in the art can assess whether the knockdown of native MHC I / II has been achieved by evaluating the mRNA level or protein level of MHC I / II, for example, by PCR-based assay or Western blotting, respectively.

[0126] In one embodiment, cells are further modified to knock out native MHC I / II.Methods for modifying cells to knock out native MHC I / II are known in the art, and include, for example, expressing RNA interference (such as short hairpin RNA, small interfering RNA, double-stranded RNA), expressing inhibitory oligonucleotides, and nuclease-based inhibition (such as CRISPR, TALEN, meganuclease, etc.).Those skilled in the art can assess whether the knockout of native MHC I / II has been achieved by evaluating the mRNA level or protein level of MHC I / II, for example, by PCR-based assay or Western blotting, respectively.

[0127] In one aspect, described herein are engineered cells that express and / or comprise one or more of the compositions according to any of the embodiments described herein. In one aspect, described herein are engineered cells that express and / or comprise one or more multi-component CAL and / or multi-component CAR signaling polypeptides according to any of the embodiments described herein. In some embodiments, the cell is a dendritic cell, a regulatory T cell, or an effector T cell. In some embodiments, the cell is a T cell. Such cells that express and / or comprise multi-component CAL and / or multi-component CAR signaling polypeptides are referred to herein as "partial multi-component CAL" cells or "partial multi-component CAR" cells. In some embodiments, partial multi-component CAL and / or CAR cells do not express (e.g., do not comprise) nucleic acid sequences encoding multi-component CAL and / or multi-component CAR recognition polypeptides. In some embodiments, partial multi-component CAL and / or CAR cells comprise nucleic acid sequences encoding at least one multi-component CAL and / or multi-component CAR signaling polypeptide. In some embodiments, the multi-component CAL and / or multi-component CAR signaling polypeptide is present on the membrane of the cell and is expressed at a detectable level, e.g., as a transmembrane protein. In some embodiments, the signaling polypeptide further comprises a second protein interaction domain that specifically binds to the protein interaction domain of the second recognition polypeptide, e.g., the signaling polypeptide is part of an AND-gated multi-component CAL and / or an AND-gated multi-component CAR described elsewhere herein. In some embodiments, the cell can further comprise a second multi-component CAL and / or multi-component CAR signaling polypeptide, e.g., a signaling polypeptide that is part of a second multi-component CAL and / or multi-component CAR according to any of the embodiments described herein.

[0128] In one aspect, described herein is a method of killing a target cell, 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 multi-component CAR. In some embodiments, the target cell can be a diseased cell, such as an autoreactive or alloreactive T cell. In some embodiments, the target cell can be a diseased cell, such as a cancer cell (e.g., a T cell-derived or T cell precursor-derived neoplasm, hereinafter referred to as a "T cell neoplasm"). In some embodiments, the method further comprises obtaining cells (e.g., NK cells, dendritic cells, regulatory T cells, or effector T cells) from a subject, modifying the cells to contain a nucleic acid sequence encoding a signaling polypeptide of the multi-component CAL and / or multi-component CAR, and then administering the cells to the subject.

[0129] In some embodiments of any of the methods described herein, the protein interaction domain pair of the multi-component CAL and / or multi-component CAR can include a chemically induced binding domain, and the method can further include administering a compound that induces binding of the domains. In some embodiments, when one protein interaction domain is the FKBP binding domain (FRB) of mTOR and the second protein interaction domain is FK506-binding protein (FKBP), the method further includes administering tacrolimus, a rapalog, or everolimus. In some embodiments, when one protein interaction domain is cyclophilin-Fas fusion protein (CyP-Fas) and the second protein interaction domain is FK506-binding protein (FKBP), the method further includes administering FKCsA. In some embodiments, when one protein interaction domain is calcineurin A (CNA) and the second protein interaction domain is FK506-binding protein (FKBP), the method further includes administering FK506. In some embodiments, when one protein interaction domain is gibberellin-insensitive (GIA) and the second protein interaction domain is gibberellin-insensitive dwarf1 (GID1), the method further comprises administering gibberellin. In some embodiments, when one protein interaction domain is a Snap tag and the second protein interaction domain is a Halo tag, the method further comprises administering HaXS. In some embodiments, when one protein interaction domain is T14-3-3-cΔC and the other protein interaction domain is the C-terminal peptide of PMA2 (CT52), the method further comprises administering fukucoccin.

[0130] In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be modified polypeptides. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be transgenic. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be recombinant polypeptides. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be heterologous to the cell. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be heterologous to the T cell. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be heterologous to human T cells. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be exogenous to the cell. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be exogenous to the T cell. In some embodiments of any of the aspects described herein, the recognition and / or signaling polypeptides of the multicomponent CAL and / or multicomponent CAR can be exogenous to the human T cell.

[0131] It is expressly contemplated herein that each of the individual embodiments described herein can be combined, for example, in a single cell. As a non-limiting example, a single cell can include a first complete multi-component CAL and / or complete multi-component CAR and a second partial multi-component CAL and / or partial multi-component CAR, where each multi-component CAL and / or multi-component CAR can be according to any of the embodiments described herein.

[0132] In some embodiments, the methods described herein relate to CAR-immune cell therapy. In some embodiments, the methods described herein relate to CAR-immune cell therapy, such as CAR-T therapy. Standard CAR-T therapy and related therapies involve the adoptive cell transfer of immune cells (e.g., T cells) expressing a CAR that specifically binds to a target cell type (e.g., diseased cells, such as autoreactive or alloreactive T cells) to treat a subject for, for example, an autoimmune disease or condition, a T cell-mediated inflammatory or immune response, graft rejection, or GvHD.

[0133] In some embodiments, the cells administered as part of the treatment can be autologous to the subject. In some embodiments, the cells administered as part of the treatment are not autologous to the subject. In some embodiments, the cells are engineered and / or genetically modified to express the multi-component CAL and / or multi-component CAR described herein or a portion thereof. Further discussion regarding CAR-T therapy can be found, for example, 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 in its entirety.

[0134] In some embodiments, the technology described herein relates to a syringe or catheter, e.g., an organ-specific catheter (e.g., a renal catheter, a biliary catheter, a cardiac catheter, etc.), comprising a therapeutically effective amount of a composition described herein.

[0135] Treatment method In some embodiments, the methods described herein relate to the treatment or prevention of graft rejection in a subject who has received a cell transplant, tissue transplant, or organ transplant with one or more compositions, CALs, CARs, or cells described herein. In another embodiment, the methods described herein relate to the treatment or prevention of GvHD in a subject who has received a cell transplant, tissue transplant, or organ transplant with one or more compositions, CALs, CARs, or cells described herein.

[0136] As used herein, "GvHD" refers to a disease characterized by an active process in which donor cells attack the recipient's own cells. GvHD can occur shortly after transplantation, e.g., within weeks or months (acute GvHD), or it can occur much later after transplantation, e.g., at least 3-6 months (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 in 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.

[0137] A skilled clinician can identify a subject as having or at risk for GvHD. Diagnostic tests useful for identifying subjects with GvHD are known in the art and will vary based on the type of transplant the subject has received. A diagnosis of GvHD is made, for example, by physical examination for signs and symptoms of GvHD known in the art, blood tests for liver, gallbladder, kidney, and hematopoietic cell dysfunction, histological analysis of biopsies obtained from affected organs, and radiological imaging of affected organs. In one embodiment, the method further comprises administering at least a second therapeutic agent. In one embodiment, the composition, CAR, CAL, 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 immunoglobulin IgG1 fused to the extracellular domain of CTLA-4. Abatacept is currently approved by the FDA for the treatment of rheumatoid arthritis. Belatacept, which differs from abatacept by just two amino acids, is an immunosuppressant intended to prevent rejection after kidney transplants.

[0138] In one embodiment, the transplant is a revascularized composite allogeneic transplant (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 transplantation, such as 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 allogeneic transplant (e.g., transplant of allogeneic tissue), any xenogeneic transplant (e.g., transplant of xenogeneic tissue), or any autologous transplant (e.g., transplant of tissue). A skilled practitioner will be able to use standard surgical protocols to perform the transplant or identify the subject who will receive the transplant.

[0139] In some embodiments, the methods described herein relate to the treatment or prevention of autoimmune diseases or conditions, or type I-IV hypersensitivity reactions, or immune responses to therapeutic foreign proteins / molecules, or T cell-mediated inflammation or immune responses, using the compositions, CARs, CALs, or cells described herein. Physicians can use modern methods for diagnosing autoimmune diseases to identify subjects with autoimmune diseases. Symptoms and / or complications of autoimmune diseases that characterize these conditions and aid in diagnosis are well known in the art and include, but are not limited to, fatigue, muscle pain, swelling and redness, mild fever, numbness or tingling in the hands and feet, hair loss, and / or skin rash. Tests that can aid in the diagnosis of autoimmune diseases include, but are not limited to, blood counts and antinuclear antibody (ANA) tests. Having a family history of autoimmune disease or risk factors for autoimmune disease (e.g., gender, age, ethnicity, and environmental agents such as procainamide, hydralazine, mercury, gold, or silver) can also aid in determining whether a subject is likely to have an autoimmune disease or in making a diagnosis of an autoimmune disease.

[0140] As used herein, the term "autoimmune disease," "autoimmune condition," or "autoimmune disease or disorder" refers to a disease or disorder arising from and directed against an individual's own tissues, or a co-segregate or manifestation thereof, or a condition resulting therefrom.

[0141] Autoimmune-related diseases and disorders result from the body's hypersensitivity and / or abnormal immune response to substances (autoantigens) and tissues normally present in the body, also known as self- or autologous substances. This dysregulated inflammatory response leads to an exaggerated response by macrophages, granulocytes, lymphocytes, and / or T lymphocytes, which leads to abnormal tissue damage and cell death. The subsequent loss of function is associated with inflammatory tissue damage.

[0142] As used herein, an autoantigen is an endogenous protein or fragment thereof that is involved in or induces the pathological immune response. An autoantigen can be any substance or portion thereof normally found in a mammal that is the primary (or primary or secondary) target of attack by the immune system in an autoimmune disease. The term also encompasses antigenic substances that, when administered to a mammal, induce a condition characterized by an autoimmune disease. In addition, the term encompasses peptide subclasses that consist essentially of immunodominant epitopes or immunodominant epitope regions of an autoantigen. An immunodominant epitope or immunodominant region in an induced autoimmune condition is a fragment of an autoantigen that can be used in place of the entire autoantigen to induce disease. In humans affected by autoimmune disease, an immunodominant epitope or immunodominant region is a fragment of an antigen that is specific to the tissue or organ under autoimmune attack and is recognized by a significant portion (e.g., a large number, though not necessarily the absolute majority) of autoimmune-attacking T cells.

[0143] Autoantigens known to be associated with autoimmune diseases include myelin proteins and demyelinating diseases such as multiple sclerosis and experimental autoimmune myelitis; collagen and rheumatoid arthritis; insulin, proinsulin, glutamic acid decarboxylase 65 (GAD65); and pancreatic islet cell antigen (ICA512; ICA12) in insulin-dependent diabetes.

[0144] A common feature of several autoimmune-related diseases and autoimmune-related inflammatory conditions is the involvement of proinflammatory 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), gamma interferon, tumor nephropathy (TNFα), and IL-12. In some embodiments, cytokines characterized as Th1-type include interleukin 2 (IL-2), interferon gamma, and TNFα. Such proinflammatory cytokines act to stimulate immune responses, which often result in the destruction of autologous tissue. Cytokines associated with suppressing T cell responses are Th2-type, including IL-10, IL-4, and TGF-β. It has been found that Th1-type and Th2-type T cells can use the same antigen receptor in response to immunogens, with the former generating stimulatory responses and the latter generating inhibitory responses.

[0145] T cell mediated inflammation or a T cell mediated immune response is an inflammation and / or immune response in which T cells and / or T cell activity contribute to or cause the inflammatory / immune response.

[0146] As used herein, "inflammation" refers to a complex biological response to harmful stimuli, such as pathogens, damaged cells, or irritants. Inflammation is a defensive attempt by an organism to eliminate the harmful stimulus and initiate the tissue healing process. Therefore, the term "inflammation" includes any cellular process resulting from the action of proinflammatory cytokines, the production of inflammatory mediators, and / or the associated downstream cellular events resulting from the action of the cytokines so produced, such as fever, fluid retention, swelling, abscess formation, and cell death. Inflammation can encompass both acute responses (i.e., responses characterized by an active inflammatory process) and chronic responses (i.e., responses characterized by slow progression and the formation of new connective tissue). Acute and chronic inflammation can be distinguished by the cell types involved. Acute inflammation often involves polymorphonuclear neutrophils, whereas chronic inflammation is typically characterized by lymphohistiocytic and / or granulomatous responses. An inflammatory condition is any disease state characterized by inflamed tissue (e.g., infiltration of leukocytes such as lymphocytes, neutrophils, macrophages, eosinophils, mast cells, basophils, and dendritic cells) or an inflammatory process that causes or contributes to the abnormal clinical and histological features of the disease state.

[0147] As used herein, an "immune response" refers to a response by a cell of the immune system, e.g., a B cell, a T cell (CD4 or CD8), a regulatory T cell, an antigen-presenting cell, a dendritic cell, a monocyte, a macrophage, a NKT cell, a NK cell, a basophil, an eosinophil, or a neutrophil, to a stimulus (e.g., a disease, an antigen, or, in the case of, e.g., an autoimmune disease, a healthy cell). In some embodiments of the aspects described herein, the immune response is a T cell response, e.g., 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, depending on the nature of the immune cell causing the response. Stimulation of an immune response refers to induction or increase of an immune response. Suppression of an immune response refers to elimination or reduction of an immune response.

[0148] 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 major histocompatibility complex (MHC) class I or class II molecules, CD1, or other non-classical MHC-like molecules. This activates antigen-specific CD4+ T helper cells or CD8+ cytotoxic lymphocytes ("CTLs"). CTLs have specificity for peptide antigens presented in association with proteins encoded by classical or non-classical MHC and expressed on the surface of cells. CTLs help induce and promote the intracellular destruction of intracellular microorganisms or the lysis of cells infected with such microorganisms. Another aspect of cellular immunity involves antigen-specific responses by helper T cells. Helper T cells act to stimulate the function and help focus the activity of nonspecific effector cells against cells presenting peptides or other antigens in association with classical or non-classical MHC molecules on their surface. "Cell-mediated immune response" also refers to the production of cytokines, chemokines, and other similar molecules produced by activated T cells and / or other leukocytes, including those derived from CD4+ T cells and CD8+ T cells. Stimulation of a cell-mediated immunological response can be determined by several assays, such as lymphoproliferation (lymphocyte activation) assays, CTL cytotoxicity assays, assaying T lymphocytes against an antigen in a sensitized subject, or measuring cytokine production by T cells in response to antigen restimulation. Such assays are well known in the art. See, for example, Erickson et al. (1993) J. Immunol. 1 51:4189-4199 and Doe et al. (1994) Eur. J. Immunol. 24:2369-2376.

[0149] In some embodiments, the T cell-mediated immune response is a response to a drug administered to a subject.It is contemplated herein that the technology of the present invention can be used to deplete anti-drug specific T cells to prevent immune responses to administered biologics, cell therapy, and / or gene therapy.For example, the methods and compositions described herein can be used to prevent or treat anti-AAV and anti-transgene immune responses to administered adeno-associated virus (AAV) gene therapy, to prevent or treat immune responses to genome editing agents such as CRISPR / Cas9, transcription activator-like effector nucleases (TALENs) or zinc finger nucleases (ZFNs), and to prevent or treat immune responses to enzyme replacement therapy such as recombinant human acid α-glucosidase (Pompe disease), α-L-iduronidase (mucopolysaccharidosis I), and α-galactosidase (Fabry disease).

[0150] In one embodiment of any one of the described methods, the autoimmune disorder is thyroiditis, type 1 diabetes, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barré syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis, e.g., acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immune-mediated arthritis, chronic inflammatory arthritis, osteoarthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post-treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, spondyloarthritis ... and juvenile-onset rheumatoid arthritis, chronic progressive arthritis, osteoarthritis, primary chronic polyarthritis, reactive arthritis, and ankylosing spondylitis), relapsing arthritis, inflammatory hyperproliferative skin diseases, psoriasis, such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and nail psoriasis, atopy including atopic diseases, such as hay fever and Job's syndrome, dermatitis, such as contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, nonspecific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper I gM syndrome, allergic intraocular inflammatory disease, urticaria, e.g. chronic allergic urticaria and chronic idiopathic urticaria, e.g. chronic autoimmune urticaria, myositis, polymyositis / dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic sclerosis), sclerosis, e.g. systemic sclerosis, multiple sclerosis (MS), e.g. optic-spinal MS, primary progressive MS (PPMS), and relapsing-remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, disseminated sclerosis, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (e.g. Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis (e.g., ulcerative colitis, ulcerative colitis, microscopic colitis, collagenous colitis, polypoid colitis, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), enteritis, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome (e.g., adult or acute respiratory distress syndrome (ARDS)), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, autoimmune blood disorders, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as seen in meningitis, herpes gestationis, pemphigoid of gestationis,scrotal pruritus, autoimmune premature ovarian failure, sudden hearing loss due to autoimmune conditions, 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, non-granulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome, such as chronic or acute glomerulonephritis, e.g., rheumatoid arthritis ... secondary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membranoproliferative or membranoproliferative GN (MPGN) including types I and II, and rapidly progressive GN, proliferative nephritis, autoimmune polyendocrine deficiency, balanitis such as plasma cell circumscribed balanitis, balanoposthitis, erythema annulare centrifugally, erythema dyschromatosis perstans, erythema multiforme, granuloma annulare, lichen sclerosus and atrophicus, lichen simplex chronicus, lichen spinous, lichen planus, ichthyosis lamellar, epidermolytic keratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, Eczema, e.g., allergic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and bullous palmoplantar eczema; asthma, e.g., bronchial asthma, bronchial asthma, and autoimmune asthma; conditions involving T cell infiltration and chronic inflammatory responses; immune responses to foreign antigens such as fetal ABO blood group during pregnancy; chronic pulmonary inflammatory diseases; autoimmune myocarditis; leukocyte adhesion deficiency; lupus, e.g., lupus nephritis, lupus encephalitis, childhood lupus, non-renal lupus, extrarenal lupus, discoid lupus, and discoid lupus erythematosus; lupus alopecia; systemic lupus erythematosus (S LE), such as cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and disseminated lupus erythematosus, juvenile-onset (type I) diabetes, such as insulin-dependent diabetes mellitus (IDDM) of childhood, adult-onset diabetes mellitus (type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic aortopathy, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, sarcoidosis, granulomatous diseases, such as lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitis, such as vasculitis,Necrotizing vasculitis such as large-vessel vasculitis (including polymyalgia rheumatica and giant cell (Takayasu) arteritis), medium-sized vasculitis (including Kawasaki disease and polyarteritis nodosa / periarteritis nodosa), microscopic polyarteritis, immune-mediated vasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, systemic necrotizing vasculitis, and ANCA-associated vasculitis such as Churg-Strauss vasculitis or Churg-Strauss syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs-positive anemia, Diamond-Blackfan anemia, hemolytic anemia, or immune hemolytic anemias such as autoimmune hemolytic anemia (AIHA), pernicious anemia (pernicious anemia), Addison's disease, pure red cell anemia or pure red cell aplasia (PRCA), factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases with leukocyte leakage, CNS inflammatory disorders, multiple organ injury syndromes such as those secondary to sepsis, trauma or hemorrhage, antigen-antibody complex mediated diseases, antiglomerular basement membrane disease, antiphospholipid syndrome, allergic neuritis, Behcet's disease / syndrome, Castleman syndrome, Goodpasture's syndrome, Raynaud's syndrome, Sjögren's syndrome, Stevens-Johnson syndrome, pemphigoid, e.g., bullous pemphigoid and cutaneous pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, mucous membrane pemphigoid, and pemphigus erythematous), autoimmune polyendocrinopathy, Reiter's disease or Reiter's syndrome, immune complex diseases such as immune complex nephritis, antibody-related nephritis, chronic neuropathy such as polyneuropathy, IgM polyneuropathy or IgM-related neuropathy, and autoimmune or immune-mediated thrombocytopenia, e.g., chronic ITP or idiopathic thrombocytopenic purpura (ITP) including acute ITP, scleritis, e.g. idiopathic keratoscleritis, episcleritis, autoimmune diseases of the testes and ovaries, e.g. autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis, e.g. autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis) or subacute thyroiditis, idiopathic hypothyroidism, Graves' disease, polyglandular syndromes, e.g. autoimmune polyglandular syndrome (or polyendocrinopathy syndrome), paraneoplastic syndromes including neurological paraneoplastic syndromes,Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis, e.g. allergic encephalomyelitis or allergic encephalomyelitis 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 syndrome, autoimmune hepatitis, lupoid hepatitis, giant cell hepatitis, autoimmune chronic active hepatitis, lymphocytic interstitial pneumonia (LIP), bronchiolitis obliterans (non-transplant) vs. NSIP, Guillain-Barré syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis, e.g. primary biliary cirrhosis and pulmonary cirrhosis, autoimmune enteropathy syndrome, celiac disease or celiac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, muscle atrophy Autoimmune lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear diseases such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsing or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome / non-syphilitic interstitial keratitis, Bell's palsy, Sweet's disease / syndrome, autoimmune rosacea, shingles-associated pain, amyloidosis, non-cancerous lymphocytosis, monoclonal antibody tests primary lymphocytosis including central B-cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathies, paraneoplastic syndromes, channelopathies including CNS channelopathies, autism, inflammatory myopathies, focal or segmental glomerulosclerosis or focal segmental glomerulosclerosis (FSGS), endocrine ophthalmopathy, uveitis, chorioretinitis, autoimmune liver disease, fibromyalgia, polyendocrine deficiency, Schmidt syndrome, adrenalitis, gastrotrophy, presenile dementia, autoimmune demyelinating diseases and demyelinating diseases such as chronic inflammatory demyelinating polyneuropathy, Dressler syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility,for example due to antisperm antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortions, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, fancier's lung, allergic granulomatous vasculitis, benign lymphocytic vasculitis, Alport's syndrome, alveolitis, e.g. allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reactions, Samter's syndrome, Kaplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial pulmonary fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum elevata, erythroblastosis fetalis, eosinophilic fasciitis, Charmant's syndrome group, Felty's syndrome, cyclitis, e.g., chronic cyclitis, heterochromic iritis, iridocyclitis (acute or chronic), or Fuchs' cyclitis, Henoch-Schönlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndrome, Evans' syndrome, autoimmune dysgonadism, Sydenham's chorea, post-streptococcal nephritis, thromboangiitis obliterans, thyrotoxicosis, spinal cord fistula, choroiditis, giant cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplanted organ reperfusion, autoimmune retinopathy, aphthous ulcers, Aphthous stomatitis, arteriosclerotic disorders, azoospermia, autoimmune hemolysis, Beck's disease, allergic enterocolitis, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, rheumatic fever, Hammann-Rich disease, sensorineural hearing loss, focal ileitis, leukopenia, transverse myelitis, primary idiopathic myxedema, sympathetic ophthalmia, acute polyradiculitis, pyoderma gangrenosum, acquired splenic atrophy, vitiligo, toxic shock syndrome, conditions involving T-cell infiltration, leukocyte adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, diseases involving leukocyte leakage, multiple organ injury syndrome, Antigen-antibody complex mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathy, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine deficiency, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid sinusitis, frontal sinusitis, maxillary sinusitis or sphenoid sinusitis, eosinophil-related disorders such as eosinophilia, eosinophilic pulmonary infiltrates, eosinophilic myalgia syndrome, Löffler's syndrome,The autoimmune disorder is selected from the group consisting of chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, eosinophil-containing granuloma, seronegative spondyloarthritis, polyglandular autoimmune disease, sclerosing cholangitis, sclera, episclera, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia-telangiectasia syndrome, vascular ectasia, autoimmune disorders associated with connective tissue disease, rheumatism, allergic hypersensitivity disorders, glomerulonephritis, reperfusion injury, ischemia-reperfusion injury, lymphomatous tracheobronchitis, inflammatory skin diseases, skin diseases with an acute inflammatory component, and autoimmune uveoretinitis (AUR). The infectious disease or autoimmune condition or T cell mediated inflammation can be neurodegeneration, for example Alzheimer's disease or Parkinson's disease.

[0151] In some embodiments of any aspect, 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.

[0152] As used herein, a "malignant T-cell condition" refers to a condition in which T cells exhibit one or more of the following: uncontrolled growth (i.e., division beyond normal limits), infiltration (i.e., invasion and destruction of adjacent tissues), and metastasis (i.e., spread to other parts of the body via the lymph or blood). Non-limiting examples of malignant T-cell conditions include T-cell cancer, lymphoma, leukemia, T-cell acute lymphoblastic leukemia, and T-cell lymphoblastic lymphoma.

[0153] The compositions and methods described herein can be administered to a subject to treat or prevent an autoimmune disease or condition, a T cell-mediated inflammation or immune response, or transplant rejection. In some embodiments, the methods described herein include administering to a subject an effective amount of a composition, CAL, CAR, or cell described herein to alleviate symptoms of an autoimmune disease or condition, a T cell-mediated inflammation or immune response, or transplant rejection. As used herein, "alleviating symptoms" refers to ameliorating any condition or symptom associated with a disease or condition. Such a reduction is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, 99%, or greater compared to an equivalent untreated control, as measured by any standard technique. Various means for administering the compositions described herein to a subject are known to those skilled in the art. Such methods may include, but are not limited to, oral, parenteral, intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, cutaneous, topical, injection, or intratumoral administration. Administration may be local or systemic.

[0154] The administration of the compositions contemplated herein can be carried out in any convenient manner, such as by aerosol inhalation, injection, ingestion, infusion, implantation, or transplantation. In a preferred embodiment, the composition is administered parenterally. As used herein, the phrases "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravascular, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intratumoral, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intrathecal, 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.

[0155] Pharmaceutical compositions comprising the cells described herein, e.g., T cells or CAL cells or CAR cells, can be administered in a 10 2 ~1010 cells / kg body weight, preferably 10 5 ~10 6 It can generally be stated that the composition may be administered at a dosage of cells / kg body weight (including all integer values ​​within these ranges). The number of cells will depend on the end use of the composition and the type of cells contained therein. For the uses provided herein, the cells are generally in a volume of 1 liter or less, which can be 500 mL or less, or even 250 mL or less, or 100 mL or less. Thus, the desired cell density is typically 10 6 cells / ml, typically >10 7 cells / ml, typically >10 8 cells / ml or more. A clinically relevant number of immune cells is a cumulative total of 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 or 10 12 In some aspects of the invention, the number of cells administered can be divided into multiple infusions of 10 or more cells, particularly since all of the infused cells will be redirected to a particular or specific target antigen. 6 Pieces / kilogram (10 per patient) 6 ~10 11 The dose can be as low as 100 mg / kg or less, ranging from 100 mg / kg to 100 mg / kg (e.g., 100 mg / kg or 100 mg / kg). The CAL- and / or CAR-expressing cell compositions can be administered multiple times at dosages within these ranges. The cells can be allogeneic, syngeneic, xenogeneic, or autologous to the patient undergoing treatment. If desired, the treatment can also include administration of a mitogen (e.g., PHA) or lymphokines, cytokines, and / or chemokines (e.g., IFN-γ, IL-2, IL-12, TNF-α, IL-18 and TNF-β, GM-CSF, IL-4, IL-13, Flt3-L, RANTES, MIP1α, etc.) as described herein to enhance the induction of an immune response.

[0156] In some embodiments, the dosage is about 1 x 10 per kg of body weight. 5 cells ~ approx. 1 x 108 In some embodiments, the dosage is about 1 x 10 cells per kg of body weight. 6 cells ~ approx. 1 x 10 7 In some embodiments, the dosage is about 1 x 10 cells per kg of body weight. 6 The cells can be cells. In some embodiments, the administration of the cells can be performed once. In some embodiments, the administration of the cells can be repeated, for example, once, twice or more times. In some embodiments, the administration of the cells can be performed daily, weekly, or monthly.

[0157] The dosage range of an agent, e.g., a CAL, CAR, cell, or composition described herein, depends on potency and includes an amount sufficient to produce the desired effect, e.g., prevention of transplant rejection, reduction of inflammation, etc. The dosage should not be so high as to cause unacceptable adverse side effects. Generally, dosages vary depending on the age, condition, and sex of the patient and can be determined by one of ordinary skill in the art. Dosages may also be adjusted by individual physicians if any complications are present. In some embodiments, dosages range from 0.001 mg / kg body weight to 0.5 mg / kg body weight. In some embodiments, the dosage range is from 5 μg / kg body weight to 100 μg / kg body weight. Alternatively, the dosage range can be titrated to maintain serum levels between 1 μg / mL and 1000 μg / mL. For systemic administration, a subject can be administered a therapeutic amount such as, for example, 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.

[0158] The administration of the doses described 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 described above are administered daily for several weeks or months.The duration of treatment depends on the clinical progress of the subject and their response to treatment.

[0159] In some embodiments, the dose can be 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 about 100 mg / kg. 2 ~about 700mg / m 2 In some embodiments, the dose can be about 250 mg / m 2 In some embodiments, the dose can be about 375 mg / m 2 In some embodiments, the dose can be about 400 mg / m 2 In some embodiments, the dose can be about 500 mg / m 2 It can be.

[0160] In some embodiments, the dose can be administered intravenously. In some embodiments, the intravenous administration can be an infusion administered over a period of about 10 minutes to about 3 hours. In some embodiments, the intravenous administration can be an infusion administered over a period of about 30 minutes to about 90 minutes.

[0161] In some embodiments, the dose can be administered approximately once a week. In some embodiments, the dose can be administered once a week. In some embodiments, the dose can be administered once a week for about 12 weeks to about 18 weeks. In some embodiments, the dose can be administered about every two weeks. In some embodiments, the dose can be administered about every three weeks. In some embodiments, the dose can be about 2 mg / kg to about 15 mg / kg administered about every two weeks. In some embodiments, the dose can be about 2 mg / kg to about 15 mg / kg administered about every three weeks. In some embodiments, the dose can be about 2 mg / kg to about 15 mg / kg administered intravenously about every two weeks. In some embodiments, the dose can be about 2 mg / kg to about 15 mg / kg administered intravenously about every three weeks. In some embodiments, the dose can be about 200 mg / m2 to about 400 mg / m2 administered intravenously about every week. In some embodiments, the dose can be about 200 mg / m2 to about 400 mg / m2 administered intravenously about every two weeks. In some embodiments, the dose can be about 200 mg / m2 to about 400 mg / m2 administered intravenously about every three weeks. In some embodiments, about two to about ten total doses are administered. In some embodiments, four total doses are administered. In some embodiments, five total doses are administered. In some embodiments, six total doses are administered. In some embodiments, seven total doses are administered. In some embodiments, eight total doses are administered. In some embodiments, the administration occurs over a total period of about four to about twelve weeks. In some embodiments, the administration occurs over a total period of about six weeks. In some embodiments, the administration occurs over a total period of about eight weeks. In some embodiments, the administration occurs over a total period of about twelve weeks. In some embodiments, the initial dose is about 1.5 to about 2.5 times greater than the subsequent doses.

[0162] In some embodiments, the dose can be 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.

[0163] A therapeutically effective amount is the amount of an agent sufficient to produce a statistically significant and measurable change in or prevent the development of an autoimmune disease or condition, T cell-mediated inflammatory or immune response, graft rejection, or GvHD. Such effective amounts can be estimated from clinical trials and animal studies.

[0164] The active substance can be administered intravenously by injection or by slow infusion over time.For example, if the appropriate formulation is given for a given route, the active substance useful in the methods and compositions described herein can be administered intravenously or intranasally, by inhalation, intraperitoneally, intramuscularly, subcutaneously, intracavity, and if desired, can be delivered by peristaltic means or other means known to those skilled in the art.The compound used herein is preferably administered orally, intravenously or intramuscularly.Local administration, for example, direct local administration to organ transplantation site or tissue transplantation site, is also particularly considered.

[0165] The therapeutic composition that contains at least one active substance can be conventionally administered, for example, in unit dose.When the term " unit dose " is used in relation to therapeutic composition, it refers to a physically discrete unit that serves as a unit dosage for subject, and each unit contains a predetermined amount of active substance calculated to produce desired therapeutic effect, together with necessary physiologically acceptable diluent, i.e., carrier or vehicle.

[0166] The compositions are administered in a manner compatible with the dosage formulation, and in such amount as is therapeutically effective. The amount and timing to be administered will depend on the subject to be treated, capacity of the subject's system to utilize the active ingredient, and degree of therapeutic effect desired.

[0167] In embodiments in which a subject is administered a partial multi-component CAL and / or CAR cells and a recognition polypeptide, the partial multi-component CAL and / or CAR cells and the recognition polypeptide can be administered together or separately. In embodiments in which a subject is administered a partial multi-component CAL and / or CAR cells and a recognition polypeptide separately, each of the compositions can be administered separately according to any of the dosages and routes / routines of administration described herein.

[0168] The exact amount of active ingredient that needs to be administered depends on the judgment of the practitioner and is specific to each individual.However, the appropriate dosage range for systemic application is disclosed herein, and it depends on the route of administration.Suitable administration regimen also varies, but typically includes initial administration, followed by repeated administration at intervals of 1 hour or more by subsequent injection or other administration.Alternatively, continuous intravenous infusion sufficient to maintain blood concentration within the range specified for in vivo treatment is also conceivable.

[0169] In some embodiments, the method further comprises administering the composition, CAL or CAR, or cells described herein together with one or more additional autoimmune, GvHD, or transplant rejection agents, biologics, drugs, or treatments as part of a combination therapy. Exemplary treatments for graft rejection or GvHD include immunosuppressants such as 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); chemotherapy drugs such as methotrexate (Trexall, Rasuvo, Rheumatrex, and Otrexup (PF)), antithymocyte globulin (Atgam, Thymoglobulin); steroids such as prednisone (Deltasone, Rayos, and Prednisone) antifungal agents such as posaconazole (Noxafil); antiviral agents such as acyclovir (Zovirax and Sitavig), valacyclovir (Valtrex); and antibiotics such as sulfamethoxazole / trimethoprim (Bactrim, Sulfatrim, and Bactrim DS); protease inhibitors such as alpha 1-proteinase inhibitors (Zemaira); extracorporeal photochemotherapy; monoclonal antibodies (daclizumab (Zinbryta), basiliximab (Simulect)), brentuximab vedotin (Adcetris), alemtuzumab (Campath, Lemtrada), tocilizumab (Actemra); and mesenchymal stromal cell infusions.

[0170] Exemplary treatments for autoimmune diseases include 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 supplements, e.g., glucose tablets; and hormones, e.g., glucagon (GlucaGen and Glucagon Emergency Kit (Human)), antidiabetic drugs (metformin (D-Care DM2, Fortamet, Glucophage, Glucophage)), and the like. XR, Glumetza, Riomet), glucagon-like peptide-1 (GLP-1) receptor agonists (liraglutide (Saxenda; Victoza) or semaglutide (Ozempic) or sodium-glucose cotransporter 2 (SGLT2) inhibitors: empagliflozin (Jardiance), canagliflozin (Invokana); sulfonylureas: glipizide (GlipiZIDE XL, Glucotrol, Glucotrol XL); meglitinide analogs: repaglinide (Prandin); thiazolidinediones: pioglitazone (Actos); dipeptidyl peptidase-4 (DPP-4) inhibitors: sitagliptin (Januvia), saxagliptin (Onglyza), linagliptin (Tradjenta), alogliptin (Nesina).

[0171] A skilled clinician can determine the efficacy of a given treatment, for example, for an autoimmune disease or condition, a T-cell-mediated inflammatory or immune response, a transplant rejection, or GvHD. A treatment is considered "effective" within the meaning of the present specification if, after treatment with an agent described herein, any one or all of the signs or symptoms are beneficially altered, or other clinically acceptable symptoms are improved, for example, by at least 10%, or even ameliorated. Efficacy can also be measured by the absence of deterioration in an individual (i.e., cessation of disease progression) as assessed by the need for hospitalization or medical intervention. Methods for measuring these indicators are known to those skilled in the art and / or are described herein.

[0172] An effective amount for treating a disease means an amount sufficient to provide effective treatment for the disease, as defined herein, when administered to a mammal in need thereof. The efficacy of an agent can be determined, for example, by assessing physical indicators of autoimmune disease (e.g., ANA results), T cell-mediated inflammatory or immune response, malignant T cell conditions, graft rejection (e.g., high fever, tenderness at the transplant site, etc.), or GvHD (e.g., redness, pain, or other symptoms at the transplant site). Effective amounts, toxicity, and therapeutic efficacy can be determined in cell cultures or experimental animals using standard pharmaceutical procedures, for example, to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). Dosages can vary depending on the dosage form used and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the ratio LD50 / ED50. Compositions and methods exhibiting large therapeutic indices are preferred. The therapeutically effective dose can be initially estimated from cell culture assays. Dosages can also be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of active ingredient that achieves 50% maximum inhibition of symptoms) determined in cell culture or a suitable animal model. Plasma levels can be measured, for example, by immunoassays, various DNA detection techniques, or high-performance liquid chromatography. The effect of any particular dosage can be monitored by appropriate bioassays, such as assays to evaluate post-transplant response, inflammation levels, ANA measurements, etc. A physician can determine the dosage and adjust it, if necessary, to the observed therapeutic effect.

[0173] Efficacy can also be measured by the absence of deterioration of an individual (i.e., halting of disease progression) as assessed by the need for hospitalization or medical intervention. Methods for 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 animal (some non-limiting examples of which include humans or animals), including, for example: (1) inhibiting the disease, e.g., preventing the worsening of symptoms (e.g., pain or inflammation), or (2) reducing the severity of the disease, e.g., causing regression of symptoms. An effective amount for treating a disease means an amount sufficient to result in effective treatment of the disease as defined herein when administered to a subject in need thereof. The efficacy of an agent can be determined by assessing physical indicators of a condition or desired response (e.g., reduced inflammation, etc.). It is well within the capabilities of one of skill in the art to monitor the efficacy of administration and / or treatment by measuring any one or any combination of such parameters. Efficacy can be assessed in animal models of the conditions described herein, e.g., animal models for the treatment of autoimmune disease, transplant rejection, or GVHD. When experimental animal models are used, efficacy of treatment is demonstrated when a statistically significant change in a marker, such as inflammation, is observed.

[0174] 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 multi-component CAR described herein (or a portion thereof, or cells comprising a CAL and / or CAR, or a multi-component CAL and / or multi-component CAR), and optionally a pharmaceutically acceptable carrier. In some embodiments, the active ingredient of the pharmaceutical composition comprises a CAL and / or CAR or a multi-component CAL and / or multi-component CAR described herein (or a portion thereof, or cells comprising a CAL and / or CAR or a multi-component CAL and / or multi-component CAR). In some embodiments, the active ingredient of the pharmaceutical composition consists essentially of a CAL and / or CAR or a multi-component CAL and / or multi-component CAR described herein (or a portion thereof, or cells comprising a CAL and / or CAR or a multi-component CAL and / or multi-component CAR). In some embodiments, the active ingredient of the pharmaceutical composition consists of a CAL and / or CAR or a multi-component CAL and / or multi-component CAR described herein (or a portion thereof, or cells comprising a multi-component CAL and / or multi-component CAR).

[0175] 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 that 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 carboxymethylcellulose, methylcellulose, ethylcellulose, microcrystalline cellulose and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) lubricants, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository wax; (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. glycols; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol (PEG); (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, 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 buffer solutions; (21) polyesters, polycarbonates, and / or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids, (23) serum components, such as serum albumin, HDL, and LDL; (22) C2-C12 alcohols, such as ethanol; and (23) other non-toxic compatible substances used in pharmaceutical formulations. Wetting agents, coloring agents, release agents, coating agents, sweeteners, flavoring agents, fragrances, preservatives, and antioxidants may also be present in the formulation. The terms "excipient," "carrier," "pharmaceutically acceptable carrier," and the like are used interchangeably herein. In some embodiments, the carrier inhibits degradation of the active agents described herein.

[0176] In some embodiments, pharmaceutical compositions comprising the multi-component CAL and / or multi-component CAR (or portions thereof, or cells comprising the multi-component CAL and / or multi-component CAR) described herein can be in a parenteral dosage form. Because administration of a parenteral dosage form typically bypasses a patient's natural defenses against contaminants, the parenteral dosage form is preferably sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, ready-to-inject solutions, dry products ready to dissolve or suspend in a pharmaceutically acceptable vehicle for injection, ready-to-inject suspensions, and emulsions. In addition, controlled-release parenteral dosage forms can be prepared for administration to a patient, including, but not limited to, DUROS®-type dosage forms and dose-dumping.

[0177] Suitable vehicles that can be used to provide parenteral dosage forms of the multi-component CAL and / or multi-component CAR (or portions thereof, or cells comprising the multi-component CAL and / or multi-component CAR) disclosed herein are well known to those skilled in the art. Examples include, but are not limited to, sterile water; USP Water for Injection; saline solution; glucose solution; aqueous vehicles such as, but not limited to, sodium chloride 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 pharmaceutically acceptable salts of the active ingredients can also be incorporated into the parenteral dosage forms of the present disclosure, including conventional controlled-release parenteral dosage forms.

[0178] Pharmaceutical compositions can also be formulated for oral administration, for example, as discrete dosage forms, such as, but not limited to, tablets (including, but not limited to, scored or coated tablets), pills, caplets, capsules, chewable tablets, powder packets, cachets, lozenges, wafers, aerosol sprays, or liquids, such as, but not limited to, syrups, elixirs, solutions, or suspensions in aqueous liquids, non-aqueous liquids, oil-in-water emulsions, or water-in-oil emulsions. Such compositions contain a predetermined amount of a pharmaceutically acceptable salt of a disclosed compound and can be prepared by pharmaceutical methods well known to those skilled in the art. For a general discussion, see Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams, and Wilkins, Philadelphia, PA. (2005).

[0179] Conventional dosage forms generally provide rapid or immediate drug release from the formulation. Depending on the pharmacology and pharmacokinetics of the drug, the use of conventional dosage forms may result in large fluctuations in drug concentrations in the patient's blood and other tissues. These fluctuations can affect several parameters, such as administration frequency, onset of action, duration of efficacy, maintenance of therapeutic blood levels, toxicity, and side effects. Advantageously, controlled-release formulations can be used to control the drug's onset of action, duration of action, serum levels within the therapeutic window, and peak blood levels. In particular, controlled-release or sustained-release dosage forms or formulations can be used to ensure that maximum drug efficacy is achieved while minimizing potential side effects and safety concerns that may arise from underdosing (i.e., below the minimum therapeutic level) or exceeding the drug's toxic levels. In some embodiments, the composition can be administered as a sustained-release formulation.

[0180] Controlled-release pharmaceutical products share a common goal: improving drug therapy compared to that achieved by their non-controlled counterparts. Ideally, the use of optimally designed controlled-release preparations in medical treatments is characterized by the use of a minimal amount of active pharmaceutical ingredient and the shortest possible time to cure or manage a condition. Advantages of controlled-release formulations include: 1) extended drug activity, 2) reduced dosing frequency, 3) improved patient compliance, 4) reduced total drug use, 5) reduced local or systemic side effects, 6) minimal drug accumulation, 7) reduced blood level fluctuations, 8) improved treatment efficacy, 9) reduced enhancement or loss of drug activity, and 10) improved speed of disease or condition management. Kim, Cherng-ju, Controlled Release Dosage Form Design, 2 (Technomic Publishing, Lancaster, PA, 2000).

[0181] Most controlled-release formulations are designed to initially release an amount of drug (active ingredient) that immediately produces the desired therapeutic effect, and then gradually and continuously release another amount of drug so that that level of therapeutic or prophylactic effect is maintained over an extended period of time. To maintain this constant drug level 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.

[0182] Various known controlled-release or sustained-release dosage forms, formulations and devices can be adapted for use with the salts and compositions of the present disclosure.Examples include, but are not limited to, those described in the following U.S. Patents: U.S. Patent 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 U.S. Patents is incorporated herein by reference). These dosage forms can be used to provide slow or controlled release of one or more active ingredients, for example, using hydroxypropyl methylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems (e.g., OROS® (Alza Corporation, Mountain View, Calif., USA)), or by combining them in various ratios to obtain the desired release profile.

[0183] Described herein is a treatment called "CAL cell therapy" that attempts to help immune killer cells recognize autoreactive and alloreactive T cells. This is achieved by genetically modifying immune cells to express chimeric antigen ligands (CALs). CALs are engineered ligands in which the natural recognition moiety has been removed and replaced with a synthetic recognition moiety (including any synthetic or natural peptide-MHC complex) designed to more effectively recognize autoreactive and alloreactive T cells by highly specifically detecting the presence of T cell receptors unique to autoreactive and alloreactive T cells. These CAL immune cells are then administered to a patient. In the patient's body, these synthetic CAL ligand molecules bind to autoreactive and alloreactive T cells, and upon binding, activate killer immune cells, resulting in engineered CAL immune cells that attack pathological autoreactive and alloreactive T cells and eliminate or reduce the alloreactive or autoreactive immune response. This has particular application in autoimmune conditions, cell transplants, tissue transplants, organ transplants, and graft-versus-host disease (GvHD).

[0184] The present invention is based, in part, on the discovery that the modified polypeptides provided herein can recognize and bind to specific T cell receptors on disease-causing T cells, causing the deletion of the T cells and eliminating or reducing, for example, autoimmune diseases or conditions, T cell-mediated inflammation or immune responses, and cell transplants, tissue transplants, organ transplants, and graft-versus-host disease (GvHD). In some embodiments, the modified polypeptides are comprised of a peptide-major histocompatibility complex (pMHC) (e.g., as a monomer, oligomer, or multimer) as a recognition site for the TCR of allogeneic or autoreactive T cells. In some embodiments, the pMHC is one of the complexes described herein. See Tables 5 and 6.

[0185] In some embodiments, the modified polypeptide is composed of pMHC conjugated with FITC, PE, or other biomolecular interaction domains described herein. The modified polypeptide can also be considered as an adapter molecule (referred to herein as chimeric antigen ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomer, oligomer, or multimer) fused with a biomolecular interaction domain. The CAL technology described herein can target T cell clones in an antigen-specific manner and exhibits killing effects independent of any specific CAR construct.

[0186] In some embodiments, the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as a recognition site fused to a signaling domain derived from a T cell receptor. Also 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 UniCAL), which has a T cell signaling domain as an intracellular portion and a biomolecular interaction domain, e.g., specific for each disease state, as an extracellular domain. The second piece is an adapter molecule (referred to herein as chimeric antigen ligand (CAL)) composed of a TCR recognition domain (e.g., a peptide-HLA monomer, oligomer, or multimer) fused to a cognate biomolecular interaction domain.

[0187] In some embodiments, the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as a recognition site fused to a signaling domain derived from a T cell receptor. A split version of this CAR system is further provided herein, in which the CAR is composed of two pieces. The first piece is a universal CAR (also referred to herein as UniCAL), which has a T cell signaling domain as an intracellular portion and a biomolecular interaction domain, e.g., specific to each disease state, as an extracellular domain. The second piece is an adapter molecule (referred to herein as chimeric antigen ligand (CAL)) composed of a TCR recognition domain (e.g., a peptide-HLA monomer, oligomer, or multimer) fused to a cognate biomolecular interaction domain. Note that a protein interaction domain is one type of biomolecular interaction domain, and where one is specified herein, it can always be replaced with the other. The CAL technology described herein can antigen-specifically target T cell clones and exhibits a killing effect independent of any specific CAR construct.

[0188] Accordingly, one aspect presented herein provides a composition comprising (a) a chimeric antigen ligand or TCR recognition domain and one or both of (a) an intracellular signaling domain and (b) a first type protein interaction domain.

[0189] 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 of protein interaction domain; and (b) a signaling polypeptide comprising a second type of protein interaction domain and an intracellular signaling domain, wherein the first type and the second type of protein interaction domain specifically bind to each other.

[0190] Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain, and (b) a recognition polypeptide comprising a second recognition domain and a third type of protein interaction domain, wherein the first type and the third type of protein interaction domain specifically bind.

[0191] Another aspect provided herein provides a composition comprising: (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; (b) a signaling polypeptide comprising a second type of protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising the second recognition domain and a third type of protein interaction domain, wherein the second type of protein interaction domain and the third type of protein interaction domain compete for binding to the first type of protein interaction domain.

[0192] In one embodiment of any aspect, the third type protein interaction domain and the first type protein interaction domain have a higher affinity for each other than the second type protein interaction domain and the first type protein interaction domain.

[0193] Another aspect provided herein provides a composition comprising: (a) a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; (b) a signaling polypeptide comprising a second type of protein interaction domain, a fourth type of protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth type of protein interaction domain, wherein the first type of protein interaction domain and the second type of protein interaction domain specifically bind to each other, and the fourth type of protein interaction domain and the fifth type of protein interaction domain specifically bind to each other.

[0194] In one embodiment of any aspect, the fourth type protein interaction domain and the fifth type protein interaction domain have a weaker affinity than the second type protein interaction domain and the first type protein interaction domain.

[0195] In one embodiment of any aspect, the first polypeptide further comprises a protein interaction domain of type 6 and the recognition polypeptide further comprises a protein interaction domain of type 7, and they specifically bind to each other.

[0196] In one embodiment of any aspect, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.

[0197] In one embodiment of any aspect, the second recognition domain is specific for a target found on healthy cells and / or non-target cells and not on diseased cells and / or target cells.

[0198] A TCR recognition domain may not comprise a single polypeptide but may comprise two or more polypeptides, and may additionally comprise non-polypeptides. For example, a single MHC class II tetramer TCR recognition domain may comprise four small biotin molecules and 16 polypeptides (four peptides, four MHC class II α chains, four MHC class II β chains, and four streptavidin proteins). Other types of TCR recognition domains may additionally comprise other non-polypeptide molecules (e.g., an MHC dextramers contains a polysaccharide backbone to which the MHC is anchored). In some embodiments, the compositions described herein may comprise multiple copies or instances of a TCR recognition domain. For example, the TCR recognition domain can be a multimer or oligomer. In some embodiments, the compositions described herein may comprise multiple copies or instances of a first polypeptide 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 distinct polypeptide sequences, wherein a first polypeptide comprises at least one of said distinct polypeptide sequences of the TCR recognition domain, and wherein the first polypeptide binds to or complexes with a second or further polypeptide sequence of the TCR recognition domain to form the TCR recognition domain.

[0199] In one embodiment of any aspect, the TCR recognition domain comprises an MHC (major histocompatibility complex), an MHC-peptide complex, or an MHC-peptide fusion.

[0200] In one embodiment of any aspect, the peptide is a human, non-human, or synthetic / modified peptide. The peptide can further comprise a non-protein motif, modification, or domain, for example, the peptide can comprise glycosylation and / or lipids. In one embodiment of any aspect, the peptide is a minor histocompatibility antigen (MiHA).

[0201] In one embodiment of any aspect, the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form.

[0202] In one embodiment of any aspect, the protein interaction domain is found in the extracellular portion of the respective polypeptide.

[0203] In one embodiment of any aspect, (a) the protein interaction domain is a leucine zipper or any binding pair of the protein interaction domains collectively is a leucine zipper pair; (b) the protein interaction domain is a BZip(RR) and / or AZip(EE) or any binding pair of the protein interaction domains collectively is a BZip(RR) and AZip(EE); (c) the protein interaction domain is a PSD95-Dlg1-zo-1 (PDZ) domain; and (d) the protein interaction domain is a streptomycin-binding domain. (e) the protein interaction domain is mTOR's FKBP binding domain (FRB) and / or FK506 binding protein (FKBP), or any binding pair of the protein interaction domain is mTOR's FKBP binding domain (FRB) and FK506 binding protein (FKBP), or any binding pair of the protein interaction domain is mTOR's FKBP binding domain (FRB) and FK506 binding protein (FKBP), and (f) the protein (g) the protein interaction domain is cyclophilin-Fas fusion protein (CyP-Fas) and / or FK506 binding protein (FKBP), or any binding pair of the protein interaction domains as a whole is cyclophilin-Fas fusion protein (CyP-Fas) and FK506 binding protein (FKBP); (g) the protein interaction domain is calcineurin A (CNA) and / or FK506 binding protein (FKBP), or any binding pair of the protein interaction domains as a whole is calcineurin A (CNA). A) and FK506-binding protein (FKBP); (h) the protein interaction domain is gibberellin-insensitive (GIA) and / or gibberellin-insensitive dwarf1 (GID1), or any binding pair of the protein interaction domain as a whole is gibberellin-insensitive (GIA) and gibberellin-insensitive dwarf1 (GID1); (i) the protein interaction domain is a Snap tag and / or a Halo tag, or any binding pair of the protein interaction domain as a whole is a Snap tag and a Halo tag;(j) the protein interaction domain is T14-3-3-cΔC and / or the C-terminal peptide of PMA2 (CT52), or any binding pair of the protein interaction domain as a whole is T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52); (k) the protein interaction domain is PYL and / or ABI, or any binding pair of the protein interaction domain as a whole is PYL and ABI; (l) the protein interaction domain is a nucleotide tag and / or a zinc finger domain, or any binding pair of the protein interaction domain as a whole is a nucleotide tag and a zinc finger domain; (m) a biological (n) the molecular interaction domain is a nucleotide tag, or any biomolecular interaction domain binding pair is collectively a pair of nucleotide tags; (n) the protein interaction domain is fluorescein isothiocyanate (FITC) and / or a FITC-binding protein, or any binding pair of the protein interaction domain is collectively FITC and a FITC-binding protein; and / or (o) the protein interaction domain is (R)-phycoerythrin (R-PE / PE) and / or a R-PE / PE-binding protein, or any binding pair of the protein interaction domain is collectively FITC and a FITC-binding protein.

[0204] In one embodiment of any aspect, the nucleotide tag is a DNA tag or a dsDNA tag.

[0205] 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.

[0206] In one embodiment of any aspect, the cell comprising and / or expressing the TCR recognition domain and the intracellular signaling domain further comprises a TCR signaling-responsive promoter operably linked to a payload transgene. Such an embodiment allows transgene payload expression specifically in and / or in the vicinity of the target T cell. Suitable promoters and transgenes are known in the art, such as promoters and transgenes used in "TRUCK CAR" technology. An exemplary promoter is an 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 to target a patient's anti-cancer T cells in situ for highly specific and localized delivery of activating agents such as one or more checkpoint inhibitors (CTLA-4 [ipilimumab, tremelimumab] or PD-1 [nivolumab, pembrolizumab, pidilizumab]) and / or pro-inflammatory cytokines (e.g., IL-2, IL-12, etc.) that can expand and drive the patient's anti-cancer T cells toward an effector phenotype. This targeted delivery is expected to result in greater efficacy of checkpoint inhibitors without the systemic side effects caused by systemically administered checkpoint inhibitors and cytokines. Furthermore, the described technology allows for combination therapy with multiple different agents, as systemic side effects are minimized due to paracrine delivery.Further description of suitable promoters, payloads, and methods for making and using TRUCK technology is provided, for example, in 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 Chimielewski et al. Immunol Rev 2014 257:83-90, each of which is incorporated by reference in its entirety. In some embodiments, the cells may be allogeneic, e.g., once modified, and may be given as a pulse therapy. In some embodiments, the cells may be T cells or any other cell type described herein, e.g., NK cells. Exemplary, non-limiting pro-inflammatory cytokines include IFN, IFN-γ, TNFα, TGF-β, IL-1β, IL-6, IL-4, IL-10, IL-13, IL-2, IL-12, IL-15, and IL-27.

[0207] A promoter can be said to drive the expression or drive the transcription of a nucleic acid sequence that it regulates. The phrases "operably linked," "operably positioned," "operably linked," "under control," and "under transcriptional control" indicate that the promoter is in the correct functional location and / or orientation relative to the nucleic acid sequence that it regulates to control transcription initiation and / or expression of that sequence.

[0208] In some embodiments, the composition comprising a TCR recognition domain and a biomolecular interaction domain is a soluble molecule and / or a soluble complex.

[0209] Another aspect provided herein is a cell comprising and / or expressing any of the compositions described herein.

[0210] In one embodiment of any aspect, the TCR recognition domain comprises an MHC that is allogeneic to the cell that contains and / or expresses the composition. In one embodiment of any aspect, the TCR recognition domain comprises an MHC that is allogeneic to the cell from which the TCR is derived.

[0211] In one embodiment of any aspect, the TCR recognition domain comprises a peptide that is allogeneic to the cell that contains and / or expresses the composition. In one embodiment of any aspect, the TCR recognition domain comprises a peptide that is allogeneic to the cell from which the TCR is derived.

[0212] In one embodiment of any aspect, the cell is a dendritic cell (CAL DC), a T cell (e.g., effector, regulatory, etc.) (CAL-T), a regulatory T cell, an effector T cell, a natural killer cell (CAL NK), or any other myeloid cell. In one embodiment of any aspect, the cell is modified to express a polypeptide of the composition. In one embodiment of any aspect, the cell is modified to express a signaling polypeptide of the composition. In one embodiment of any aspect, the cell is further modified to knock out native MHC I / II. In one embodiment of any aspect, the cell is further modified to lack cell surface expression of native MHC I / II.

[0213] 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.

[0214] 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 of protein interaction domain, and a second polypeptide comprising (a) a second type of protein interaction domain, and (b) an intracellular signaling domain, wherein the first type of protein interaction domain and the second type of protein interaction domain specifically bind to each other.

[0215] Another aspect provided herein is a composition comprising a first polypeptide comprising (a) an anti-CD127 recognition domain and (b) a first type of protein interaction domain; a second polypeptide comprising (a) an anti-CD45RO recognition domain and (b) a fifth type of protein interaction domain; and a third polypeptide comprising (a) a second type and a fourth type of protein interaction domain and (b) an intracellular signaling domain, wherein the first type of protein interaction domain and the second type of protein interaction domain specifically bind to each other, and the fourth type of protein interaction domain and the fifth type of protein interaction domain specifically bind to each other.

[0216] Another aspect provided herein is a cell comprising any of the CARs described herein or any of the compositions described herein.

[0217] Another aspect provided herein is a method for preventing and / or treating an autoimmune disease or condition, or a T cell-mediated inflammatory or immune response, or a method for treating or preventing graft rejection or GvHD in a subject in need thereof, comprising administering to the subject any of the compositions and / or cells described herein. Another aspect provided herein is a method for preventing and / or treating a malignant T cell condition in a subject in need thereof, comprising administering to the subject any of the compositions and / or cells described herein.

[0218] In various embodiments of any aspect, the TCR recognition domain comprises an MHC allogeneic to the subject, an MHC autologous to the transplanted cells, a peptide allogeneic to the subject, or a peptide autologous to the transplanted cells.

[0219] In one embodiment of any aspect, the transplant is a revascularized composite allograft (VCA).

[0220] In one embodiment of any aspect, the autoimmune disease is type 1 diabetes, multiple sclerosis, rheumatoid arthritis, or scleroderma.

[0221] One aspect of the embodiments provided herein is a CAR T cell that targets CD127+ / CD45RO+ T cells. In one embodiment, the CD127+ / CD45RO+ T cells are CD127+ / CD45RO+ memory T cells. In one embodiment, the CD127+ / CD45RO+ T cells are alloreactive CD127+ / CD45RO+ T cells.

[0222] Thus, one aspect herein provides a CAR comprising (a) an anti-CD127 and / or anti-CD45RO recognition domain and (b) an intracellular signaling domain. Further provided herein is a composition comprising (a) an anti-CD127 and / or anti-CD45RO recognition domain and (b) an intracellular signaling domain.

[0223] Another aspect of the embodiments provides a composition comprising: (a) an anti-CD127 and / or anti-CD45RO recognition domain; (b) a first polypeptide comprising a first type of protein interaction domain; and (c) a second type of protein interaction domain and (d) an intracellular signaling domain, wherein the first type of protein interaction domain and the second type of protein interaction domain specifically bind to each other.

[0224] Another aspect of the embodiments provides a composition comprising: (a) an anti-CD127 recognition domain; (b) a first polypeptide comprising a first type of protein interaction domain; (c) an anti-CD45RO recognition domain; (d) a second polypeptide comprising a fifth type of protein interaction domain; (e) a second type and a fourth type of protein interaction domain; and (f) an intracellular signaling domain; wherein the first type of protein interaction domain and the second type of protein interaction domain specifically bind to each other, and the fourth type of protein interaction domain and the fifth type of protein interaction domain specifically bind to each other.

[0225] In one embodiment, the anti-CD127 recognition domain recognizes and binds to the sequence of CD127 on CD127+ cells, for example, on CD127+ / CD45RO+ T cells. As used herein, "CD127," also known as interleukin-7 receptor α (IL7Ra), ILRA, IL7RA, CDW127, or IL-7R-α, is a cell surface receptor that has been shown to be involved in V(D)J recombination during lymphocyte development. CD127 deficiency has been associated with severe combined immunodeficiency (SCID). CD127 sequences are known for several species, including 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 set forth in SEQ ID NO: 1. In some embodiments of any aspect, the CD127 polypeptide can be an ortholog, variant and / or allele of SEQ ID NO: 1. TIFF2025172886000080.tif97150

[0226] In one embodiment, the anti-CD45RO recognition domain recognizes and binds to the sequence of CD45RO on CD45RO+ cells, e.g., on CD127+ / CD45RO+ T cells. As used herein, "CD45RO," also known as PTPRC, LCA, LY5, B220, CD45, L-CA, T200, CD45R, and GP180, is a cell surface signaling molecule that has been shown to be an essential regulator of T cell and B cell antigen receptor signaling. The sequence of CD45RO is known and is known for several species, including 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 set forth in SEQ ID NO: 2. In some embodiments of any aspect, the CD45RO polypeptide can be an ortholog, variant and / or allele of SEQ ID NO:2. TIFF2025172886000081.tif13133

[0227] For convenience, the meanings of some terms and phrases used in the specification, examples, and appended claims are set forth below. Unless otherwise stated or necessitated by context, the following terms and phrases include the meanings set forth below. These definitions are provided to aid in the description of particular embodiments and are not intended to limit the claimed invention, as the scope of the present invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled 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 set forth herein, the definition set forth herein shall prevail.

[0228] For convenience, certain terms employed in the specification, examples, and appended claims of this application are collected here.

[0229] The terms "reduce," "reducing," "reduction," or "inhibiting" are all used herein to refer to a statistically significant decrease. In some embodiments, "reduce," "reduction," "reducing," or "inhibiting" typically refers to a decrease of at least 10% compared to a reference level (e.g., in the absence of a given treatment), and can include, for example, a decrease of 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 complete inhibition or complete reduction compared to a reference level. "Complete inhibition" is 100% inhibition compared to a reference level. The decrease is preferably to a level that is accepted as within the normal range for individuals without the given disorder.

[0230] The terms "increased," "increase," "enhance," or "activate" are all used herein to mean an increase by a statistically significant amount. In some embodiments, the terms "increased," "increase," "enhance," or "activate" can mean an increase of at least 10% compared to a reference level, e.g., 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 100%, or any increase between 10-100% compared to a reference level, or at least about 2-fold, at least about 3-fold, at least about 4-fold, or at least about 5-fold, or at least about 10-fold increase compared to a reference level, or any increase between 2-fold and 10-fold or more. In the context of a marker or symptom, an "increase" is a statistically significant increase in such level.

[0231] As used herein, the term "subject" refers to a human or animal. Typically, animals are vertebrates, such as pigs, primates, rodents, livestock, or game animals. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, such as rhesus monkeys. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Livestock and game animals include cattle, horses, pigs, deer, bison, water buffalo, felines, such as domestic cats, canines, such as dogs, foxes, wolves, birds, such as chickens, emus, ostriches, and fish, such as trout, catfish, and salmon. In some embodiments, the subject is a mammal, such as a primate, for example, a human. The terms "individual," "patient," and "subject" are used interchangeably herein.

[0232] Preferably, the subject is a mammal. The mammal can be, but is not limited to, a human, a non-human pig, a primate, a mouse, a rat, a dog, a cat, a horse, or a cow. Non-human mammals can be advantageously used as subjects corresponding to animal models of autoimmune diseases or conditions, T cell-mediated inflammation or immune responses, malignant T cell conditions, graft rejection, or GvHD. The subject can be male or female.

[0233] The subject can be one who has already been diagnosed with, or identified as suffering from, a condition in need of treatment (e.g., an autoimmune disease or condition, a T-cell-mediated inflammation or immune response, a malignant T-cell condition, a graft rejection, or GvHD) or one or more complications related to such a condition, and optionally is already receiving treatment for the autoimmune disease or condition, a T-cell-mediated inflammation or immune response, a malignant T-cell condition, a graft rejection, or GvHD, or one or more complications related to the autoimmune disease, graft rejection, or GvHD. Alternatively, the subject can be one who has not already been diagnosed with the autoimmune disease or condition, a T-cell-mediated inflammation or immune response, a malignant T-cell condition, a graft rejection, or GvHD, or one or more complications related to the autoimmune disease or condition, a T-cell-mediated inflammation or immune response, a malignant T-cell condition, a graft rejection, or GvHD. For example, the subject can be one who exhibits one or more risk factors for an autoimmune disease or condition, a T cell-mediated inflammation or immune response, a malignant T cell condition, graft rejection, or GvHD, or one or more complications related to an autoimmune disease or condition, a T cell-mediated inflammation or immune response, a malignant T cell condition, graft rejection, or GvHD, or a subject who does not exhibit risk factors.

[0234] A "subject in need" of treatment for a particular condition can be a subject who has the condition, e.g., an autoimmune disease or condition, a T cell-mediated inflammatory or immune response, a malignant T cell condition, a transplant rejection, or GvHD, a subject who has been diagnosed with the condition, or a subject who is at risk of developing the condition.

[0235] In some embodiments, the nucleic acid encoding the CAL, CAR, multi-component CAL and / or multi-component CAR described herein, or a portion thereof, is contained in a vector. In some aspects described herein, the nucleic acid sequence encoding the multi-component CAL and / or multi-component CAR described herein, or a portion thereof, or any module thereof, is operably linked to a vector. As used herein, the term "vector" refers to a nucleic acid construct designed for delivery to a host cell or transfer between different host cells. As used herein, a vector can be a viral vector or a non-viral vector. The term "vector" encompasses any genetic element that, when associated with the proper control elements, is capable of replication and can transfer a gene sequence into a cell. Vectors can include, but are not limited to, cloning vectors, expression vectors, plasmids, phages, transposons, cosmids, chromosomes, viruses, virions, etc.

[0236] As used herein, the term "expression vector" refers to a vector that directs the expression of RNA or polypeptides from sequences linked to transcriptional regulatory sequences on the vector. The sequences to be expressed are often, but not necessarily, heterologous to the cell. Expression vectors may contain additional elements. For example, an expression vector may have two replication systems, allowing it to be maintained in two organisms: human cells for expression and prokaryotic hosts for cloning and amplification. The term "expression" refers to the cellular processes involved in the production of RNA and proteins, and, where appropriate, protein secretion, including, but not limited to, transcription, transcript processing, translation, and protein folding, modification, and processing, as appropriate. "Expression product" includes RNA transcribed from a gene and polypeptides resulting from translation of mRNA transcribed from a gene. The term "gene" refers to a nucleic acid sequence (DNA) that is transcribed into RNA in vitro or in vivo when operably linked to appropriate regulatory sequences. In addition to the intervening sequences (introns) between coding segments (exons), genes may or may not include regions preceding and following the coding region, such as 5' untranslated (5'UTR) sequences or "leader" sequences and 3'UTR sequences or "trailers."

[0237] As used herein, the term "viral vector" refers to a nucleic acid vector construct that contains at least one element of viral origin and has the ability to be packaged into a viral vector particle.A viral vector can contain a nucleic acid encoding a CAL and / or CAR described herein, such as a multi-component CAL and / or multi-component CAR described herein, or a portion thereof, instead of a non-essential viral gene.Vector and / or particle can be used to transfer any nucleic acid into cells either in vitro or in vivo.Many forms of viral vectors are known in the art.

[0238] "Recombinant vector" refers to a vector containing a heterologous nucleic acid sequence or "transgene" that can be expressed in vivo or in transduced cells. It should be understood that in some embodiments, the vectors described herein can be combined with other appropriate compositions and treatments. In some embodiments, the vector is episomal. The use of an appropriate episomal vector provides a means of maintaining the nucleotide of interest in high copy number extrachromosomal DNA in a subject, thereby eliminating the potential effects of chromosomal integration.

[0239] 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 analogs thereof. Nucleic acids can be single-stranded or double-stranded. A single-stranded nucleic acid can be one nucleic acid strand of denatured double-stranded DNA. Alternatively, it can be a single-stranded nucleic acid that is not derived from any double-stranded DNA. In one aspect, a nucleic acid can be DNA. In another aspect, a nucleic acid can be RNA, such as 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.

[0240] As used herein, the terms "protein" and "polypeptide" are used interchangeably to designate a series of amino acid residues connected to each other by peptide bonds between the α-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 size or function. "Protein" and "polypeptide" are often used in reference to relatively large polypeptides, while the term "peptide" is often used in reference to small polypeptides, although the usage of these terms overlaps in the art. The terms "protein" and "polypeptide" are used interchangeably herein to refer to gene products 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 thereof.

[0241] As used herein, "antibody" refers to an IgG, IgM, IgA, IgD, or IgE molecule or antigen-specific antibody fragment thereof (including, but not limited to, Fab, F(ab')2, Fv, disulfide-linked Fv, scFv, single domain antibodies, closed conformation multispecific antibodies, disulfide-linked scFv, diabodies), whether isolated from serum, B-cells, hybridomas, transfectomas, yeast, or bacteria, whether from any species that naturally produces antibodies or produced by recombinant DNA technology.

[0242] As used herein, an "antigen" is a molecule bound by a binding site on an antibody agent. Typically, an antigen is bound by an antibody ligand and has the ability to generate an antibody response in vivo. An antigen can be a polypeptide, protein, nucleic acid, or other molecule, or a portion thereof. The term "antigenic determinant" refers to the epitope on an antigen that is recognized by an antigen-binding molecule, more specifically, by the antigen-binding site of the molecule.

[0243] As used herein, the term "antibody reagent" refers to a polypeptide that comprises at least one immunoglobulin variable domain or immunoglobulin variable domain sequence and specifically binds to a given antigen. An antibody reagent can comprise an antibody or a polypeptide comprising the antigen-binding domain of an antibody. In some embodiments, an antibody reagent can comprise a monoclonal antibody or a polypeptide comprising the antigen-binding domain of a monoclonal antibody. For example, an antibody can comprise 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 comprises two heavy (H) chain variable regions and two light (L) chain variable regions. The term "antibody reagent" encompasses not only intact antibodies but also antigen-binding fragments of antibodies (e.g., single-chain antibodies, Fab and sFab fragments, F(ab')2, Fd fragments, Fv fragments, scFv, and domain antibody (dAb) fragments (see, e.g., de Wildt et al., Eur J. Immunol. 1996;26(3):629-39, incorporated herein by reference in its entirety). Antibodies can have structural characteristics of IgA, IgG, IgE, IgD, IgM (and subtypes and combinations thereof). Antibodies can be from any source, including mouse, rabbit, pig, rat, and primates (human and non-human primates), as well as primatized antibodies. Antibodies also include midibodies, humanized antibodies, chimeric antibodies, and the like.

[0244] The VH and VL regions can be further subdivided into regions of hypervariability called "complementarity-determining regions" ("CDRs"), interspersed with highly conserved regions called "framework regions" ("FRs"). The extent of the framework regions and CDRs has been precisely defined (see Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, 5th ed., 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 herein by reference in their entireties). Each VH and VL typically consists of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0245] The terms "antigen-binding fragment" or "antigen-binding domain" are used interchangeably herein 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 by the term "antigen-binding fragment" of a full-length antibody include: (i) a Fab fragment, which is a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) an F(ab')2 fragment, which is a bivalent fragment containing two Fab fragments disulfide-bridged 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 antibody arm; (v) a dAb fragment consisting of the VH or VL domain (Ward et al., (1989) Nature 341:544-546, which is incorporated herein by reference in its entirety); and (vi) an isolated complementarity-determining region (CDR) that retains specific antigen-binding functionality.

[0246] As used herein, the term "specific binding" refers to a chemical interaction between two molecules, compounds, cells, and / or particles, in which a first entity binds to a second target entity with higher specificity and affinity than to a third, non-target entity. In some embodiments, specific binding can also refer to an affinity of a first entity for a second target entity that is at least 10 times, at least 50 times, at least 100 times, at least 500 times, at least 1000 times, or more than its affinity for a third, non-target entity. A reagent specific for a given target exhibits specific binding to that target under the conditions of the assay used. In some embodiments, the binding described herein is a preferential binding, for example, a binding between two molecules, compounds, cells, and / or particles, in which a first entity binds to a second target entity with at least two times higher specificity and affinity than to a third, non-target entity.

[0247] Additionally, as described herein, recombinant humanized antibodies can be further optimized for human therapeutic use to reduce potential immunogenicity while maintaining functional activity. In this context, functional activity refers to a polypeptide capable of exhibiting one or more known functional activities associated with a recombinant antibody or antibody reagent described herein. Such functional activities include, for example, the ability to bind to a target.

[0248] As used herein, "treat," "treatment," "treating," or "amelioration" refers to therapeutic treatment aimed at reversing, alleviating, ameliorating, inhibiting, slowing, or halting the progression or severity of a disease or disorder, such as a condition associated with an autoimmune disease or condition, a T-cell-mediated inflammation or immune response, a malignant T-cell condition, a graft rejection, or GvHD. The term "treat" encompasses reducing or alleviating at least one adverse effect or symptom of a condition, disease, or disorder, such as a condition associated with an autoimmune disease or condition, a T-cell-mediated inflammation or immune response, a malignant T-cell condition, a graft 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 the disease is reduced or halted. That is, "treatment" encompasses not only an improvement in symptoms or markers, but also a halt or at least a slowing of the progression or worsening of symptoms compared to that expected in the absence of treatment. Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptoms, whether detectable or undetectable, diminishment of the extent of the disease, stabilization of the disease state (i.e., not worsening), delay or slowing of disease progression, remission or palliation of the disease state (whether partial or complete), and / or reduced mortality. The term "treatment" of a disease also encompasses the alleviation of symptoms or side effects of the disease (including palliative treatment).

[0249] As used herein, the term "pharmaceutical composition" refers to an active agent combined with a pharmaceutically acceptable carrier, such as a carrier commonly used in the pharmaceutical industry. The phrase "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0250] As used herein, the term "administering" refers to introducing an agent disclosed herein, such as a CAL, CAR, composition, or cell, into a subject by a method or route that results in at least partial delivery of the agent at the desired site. Pharmaceutical compositions containing the compounds disclosed herein can be administered by any suitable route that results in effective treatment in the subject.

[0251] Immune checkpoint inhibitors inhibit one or more immune checkpoint proteins. The immune system has multiple inhibitory pathways that are essential for maintaining self-tolerance and regulating immune responses. For example, in T cells, the strength and quality of the response are initiated through antigen recognition by the T cell receptor and regulated by immune checkpoint proteins that balance costimulatory and inhibitory signals. In some embodiments of any aspect, the 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 that, when active, exerts an inhibitory effect on immune activity, such as T cell activity. Exemplary immune checkpoint proteins 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: 151889), and the like. Examples of such ligands include CD2 T cells (e.g., NCBI Gene ID: 3620), TDO (e.g., NCBI Gene ID: 6999), HVEM (e.g., NCBI Gene ID: 8764), GAL9 (e.g., NCBI Gene ID: 3965), 2B4 (a member of the CD2 molecule family, expressed on all NK cells, γδ T cells, and memory CD8+ (αβ) T cells) (e.g., NCBI Gene ID: 51744), CD160 (also known 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.

[0252] Non-limiting examples of immune checkpoint inhibitors include the following inhibitors (checkpoint target and manufacturer are listed in parentheses): 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), and IPH2101 (KIR; Innate Immunoglobulin G1). Pharma), KW-0761 (CCR4; Kyowa Kirin), CDX-1127 (CD27; CellDex), MEDI-6769 (Ox40; Medimmune), CP-870, 893 (CD40; Genentech), tremelimumab (CTLA-4; Mediimmune), 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; Mediimmune), IMP321, soluble Ig fusion protein (Brignone et al., 2007, J. Immunol. 179:4202-4211), 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) inhibitor (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. Patent 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 (10D1, also known as MDX-D010), U.S. Patent No. 7,488,888, PD-1 and PD-L1 blockers, nivolumab (MDX), as described in PCT Patent Application Publication Nos. WO03042402, WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400, and WO2011161699. 1106, BMS 936558, ONO4538), lambrolizumab (MK-3475 or SCH900475), CT-011, AMP-224, and BMS-936559 (MDX-1105-01). The above references are incorporated herein by reference in their entireties.

[0253] The terms "statistically significant" or "significantly" refer to statistical significance, for example, rejection of the null hypothesis with a p-value of less than 0.05, which generally means a difference of 2 standard deviations (2SD) or greater.

[0254] Except in the examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood to be modified in each instance by the term "about." When used in conjunction with percentages, the term "about" can mean ±1%.

[0255] As used herein, the term "comprising" is used in reference to compositions, methods, and each component thereof that is essential to the method or composition, but also allows for the inclusion of non-specified elements, whether essential or not.

[0256] The term "consisting of" refers to compositions, methods, and each component thereof described herein, excluding any element not recited in that description of that embodiment.

[0257] As used herein, the term "consisting essentially of" refers to elements required for a given embodiment. The term permits the presence of elements that do not materially affect the basic, novel, or functional characteristics of that embodiment.

[0258] The singular terms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. The abbreviation "eg" is derived from the Latin "exempli gratia," and is used herein to indicate a non-limiting example. Thus, the abbreviation "eg" is synonymous with "for example."

[0259] Unless otherwise defined herein, scientific and technical terms used in connection with this application shall have the meanings commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that the present invention is not limited to the particular methods, protocols, and reagents described herein, as such may 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 of this application. Definitions of common terms in immunology and molecular biology can be found in "The Merck Manual of Diagnosis and Therapy," 19th Edition, Merck Sharp & Dohme Corp., 2011 (ISBN 978-0-911910-19-3); "The Encyclopedia of Molecular Cell Biology and Molecular Medicine," edited by Robert S. Porter et al., Blackwell Science Ltd., 1999-2012 (ISBN 9783527600908); and "Molecular Biology and Biotechnology: a Comprehensive Desk Reference," edited by Robert A. Meyers, VCH Publishers, Inc.Published by Werner Luttmann, "Immunology," Elsevier, 2006; "Janeway's Immunobiology," edited by Kenneth Murphy, Allan Mowat, and Casey Weaver, Taylor & Francis Limited, 2014 (ISBN 0815345305, 9780815345305); "Lewin's Genes XI," Jones & Bartlett Publishers, 2014 (ISBN-1449659055); "Molecular Cloning: A Laboratory Manual," 4th ed., by Michael Richard Green and Joseph Sambrook, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA (2012) (ISBN 1936113414); "Basic Methods in Molecular Biology," Elsevier, 2014 (ISBN 1936113414); Science Publishing, Inc., New York, USA (2012) (ISBN 044460149X); “Laboratory Methods in Enzymology: DNA” edited by Jon Lorsch, Elsevier, 2013 (ISBN 0124199542); “Current Protocols in Molecular Biology” (CPMB), edited by Frederick M. Ausubel, John Wiley and Sons, 2014 (ISBN 047150338X, 9780471503385), "Current Protocols in Protein Science" (CPPS), edited by John E. Coligan, John Wiley and Sons, Inc., 2005; and "Current Protocols in Immunology" (CPI) (edited by John E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach, Warren Strobe, John Wiley) and Sons, Inc., 2003 (ISBN 0471142735, 9780471142737), the contents of which are incorporated herein by reference in their entireties.

[0260] Other terms are defined herein within the description of various aspects of the invention.

[0261] All patents and other publications referred to in this application, including literature references, issued patents, published patent applications, and co-pending patent applications, are specifically incorporated herein by reference to describe and disclose, for example, the methodologies described in such publications, which may be used in connection with the technology described herein. These publications are provided solely for the purpose of 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 reason of prior invention or for any other reason. Any statement as to the date of these documents or representation as to the contents of these documents is based on information available to the applicants and does not constitute an admission as to the correctness of the dates or contents of these documents.

[0262] The description of the embodiments of the present disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments or examples of the present disclosure have been described herein for illustrative purposes, various equivalent modifications are possible within the scope of the present disclosure, as will be apparent to those skilled in the relevant art. For example, while method steps or functions may be presented in a given order, in alternative embodiments, the functions may be performed in a different order, or may be performed substantially simultaneously. The teachings of the disclosure provided herein may be applied to other procedures or methods, as appropriate. The various embodiments described herein may be combined to provide additional embodiments. If necessary, aspects of the present disclosure may be modified to utilize the compositions, functions, and concepts of the above-mentioned references and applications to provide additional embodiments of the present disclosure. Furthermore, due to considerations of biological functional equivalence, some changes can be made to protein structures without affecting the type or amount of biological or chemical activity. These and other modifications can be made to the present disclosure in light of the detailed description. All such modifications are intended to be within the scope of the appended claims.

[0263] Specific elements of any of the foregoing embodiments can be combined with or substituted for elements of other embodiments. Furthermore, although advantages associated with certain embodiments of the present disclosure have been described in connection with those embodiments, other embodiments may also exhibit such advantages, and not all embodiments necessarily need to exhibit such advantages, to be encompassed within the scope of the present disclosure.

[0264] 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. Any statement as to the date of these documents or representation as to the contents of these documents is based on information available to applicant but does not constitute an admission as to the correctness of the dates or contents of these documents.

[0265] While preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the art that various changes, additions, substitutions, and the like may be made without departing from the spirit of the invention, and therefore are deemed to be within the scope of the invention as defined in the claims that follow. Moreover, it will be understood by those skilled in the art that, to the extent not already indicated, any one of the various embodiments described and illustrated herein may be further modified to incorporate features shown in any of the other embodiments disclosed herein.

[0266] Some aspects of the technology described herein can be defined by the following numbered items: Section 1. a. A TCR recognition domain; b. an intracellular signaling domain, and c. Type 1 protein interaction domain and one or both of A composition comprising: Section 2. a. a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; b. a signaling polypeptide comprising a second type of protein interaction domain and an intracellular signaling domain; wherein said first type and said second type protein interaction domains specifically bind to each other. Section 3. a. a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; b. a recognition polypeptide comprising a second recognition domain and a third type of protein interaction domain; wherein said first type and said third type protein interaction domains specifically bind. Section 4. a. a first polypeptide comprising a TCR recognition domain and a first type of protein interaction domain; b. a signaling polypeptide comprising a second type of protein interaction domain and an intracellular signaling domain; c. a recognition polypeptide comprising a second recognition domain and a third type of protein interaction domain; wherein the second type protein interaction domain and the third type protein interaction domain compete for binding to the first type protein interaction domain. Item 5. The composition of any one of items 3 to 4, wherein the third type protein interaction domain and the first type protein interaction domain have a higher affinity for each other than the second type protein interaction domain and the first type protein interaction domain. Section 6. a. a first polypeptide comprising a TCR recognition domain and a first type of 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; c. a recognition polypeptide comprising a second recognition domain and a fifth type protein interaction domain; A composition comprising: the first type protein interaction domain and the second type protein interaction domain specifically bind to each other; the fourth type protein interaction domain and the fifth type protein interaction domain specifically bind to each other; composition. Item 7. The composition of item 6, wherein the fourth type protein interaction domain and the fifth type protein interaction domain have weaker affinity than the second type protein interaction domain and the first type protein interaction domain. Item 8. The composition of any one of items 6 to 7, wherein the first polypeptide further comprises a protein interaction domain of type 6 and the recognition polypeptide further comprises a protein interaction domain of type 7, and they specifically bind to each other. Item 9. The composition of any one of items 3 to 8, wherein the second recognition domain is specific for a target not recognized by the TCR recognition domain. Item 10. The composition of any one of items 3 to 9, wherein the second recognition domain is specific for a target found on healthy cells and / or non-target cells and not found on diseased cells and / or target cells. Item 11. The composition of any one of the preceding items, wherein the TCR recognition domain comprises an MHC (major histocompatibility complex), an MHC-peptide complex, or an MHC-peptide fusion. Item 12. The composition of item 11, wherein the peptide is a human peptide or a non-human peptide. Item 13. The composition of any one of items 11 to 12, wherein the peptide is a minor histocompatibility antigen (MiHA). Item 14. The composition of any one of items 11 to 13, wherein the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. Item 15. The composition of any one of the preceding items, wherein the protein interaction domain is found in the extracellular portion of the respective polypeptide. Section 16. a. The protein interaction domain is a leucine zipper or any binding pair of the protein interaction domain as a whole is a leucine zipper pair; b. the protein interaction domain is a BZip(RR) and / or an AZip(EE), or any binding pair of the protein interaction domain as a whole is a BZip(RR) and an AZip(EE); c. the protein interaction domain is a PSD95-Dlg1-zo-1 (PDZ) domain; d. the protein interaction domain is streptavidin and / or streptavidin-binding protein (SBP), or any binding pair of the protein interaction domain is streptavidin and streptavidin-binding protein (SBP) as a whole; e. the protein interaction domain is the FKBP binding domain (FRB) of mTOR and / or the FK506 binding protein (FKBP), or any binding pair of the protein interaction domain is the FKBP binding domain (FRB) of mTOR and the FK506 binding protein (FKBP) as a whole; f. the protein interaction domain is a cyclophilin-Fas fusion protein (CyP-Fas) and / or an FK506 binding protein (FKBP), or any binding pair of the protein interaction domain is a cyclophilin-Fas fusion protein (CyP-Fas) and an FK506 binding protein (FKBP) as a whole; g. the protein interaction domain is calcineurin A (CNA) and / or FK506 binding protein (FKBP), or any binding pair of the protein interaction domain is calcineurin A (CNA) and FK506 binding protein (FKBP) as a whole; h. The protein interaction domain is gibberellin insensitive (GIA) and / or gibberellin insensitive dwarf1 (GID1), or any binding pair of the protein interaction domain is gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) as a whole; i. the protein interaction domain is a Snap tag and / or a Halo tag, or any binding pair of the protein interaction domain is a Snap tag and a Halo tag as a whole; j. the protein interaction domain is T14-3-3-cΔC and / or the C-terminal peptide of PMA2 (CT52), or any binding pair of the protein interaction domain is T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52) as a whole; k. the protein interaction domain is PYL and / or ABI, or any binding pair of the protein interaction domain as a whole is PYL and ABI, and / or l. The protein interaction domain is a nucleotide tag and / or a zinc finger domain, or any binding pair of the protein interaction domain is a nucleotide tag and a zinc finger domain as a whole; The composition according to any one of the preceding items. Item 17. The composition of item 16, wherein the nucleotide tag is a DNA tag or a dsDNA tag. 18. The intracellular signaling domain comprises: TCRC, FcRy, FcRp, CD3y, CD35, CD3s, CD3C, CD22, CD79a, CD79b, CD66d, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54(ICAM), CD83, CD134(OX4 0), 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 The composition of any one of the preceding items, wherein the signaling domain is from a protein selected from the group consisting of: Item 19. A cell comprising and / or expressing the composition of any one of the preceding items. 20. The cell of claim 19, wherein the TCR recognition domain comprises an MHC allogeneic to the cell. 21. The cell of claim 19, wherein the TCR recognition domain comprises a peptide allogeneic to the cell. Item 22. The cell according to any one of items 19 to 21, wherein the cell is a dendritic cell, a regulatory T cell, or an effector T cell. Item 23. The cell of any one of items 19 to 22, wherein the cell has been modified to express a polypeptide of the composition. Item 24. The cell of any one of items 19 to 22, wherein the cell has been modified to express the signaling polypeptide of the composition. Paragraph 25. The cell of any one of paragraphs 19 to 24, wherein the cell has been further modified to knock out native MHC I / II. Section 26. a. Anti-CD127 and / or anti-CD45RO recognition domains; b. Intracellular signaling domain A chimeric antigen receptor (CAR) comprising: Section 27. a. anti-CD127 and / or anti-CD45RO recognition domains, and b. Type 1 protein interaction domains a first polypeptide comprising: c. a second type of protein interaction domain, and d. Intracellular signaling domain and a second polypeptide comprising A composition comprising: the first type protein interaction domain and the second type protein interaction domain specifically bind to each other; composition. Section 28. a. an anti-CD127 recognition domain, and b. Type 1 protein interaction domains a first polypeptide comprising: c. an anti-CD45RO recognition domain, and d. Type 5 protein interaction domains and a second polypeptide comprising: e. Type 2 and Type 4 protein interaction domains, and f. Intracellular signaling domain and a third polypeptide comprising A composition comprising: the first type protein interaction domain and the second type protein interaction domain specifically bind to each other; the fourth type protein interaction domain and the fifth type protein interaction domain specifically bind to each other; composition. Paragraph 29. A cell comprising the CAR or composition described in any one of paragraphs 26 to 28. Section 30. A method for 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 the composition and / or cells described in any one of the preceding sections. 31. The method of claim 30, wherein the TCR recognition domain comprises an MHC allogeneic to the subject. Item 32. The method of item 30, wherein the TCR recognition domain comprises an MHC autologous to the transplanted cells. 33. The method of claim 30, wherein the TCR recognition domain comprises a peptide allogeneic to the subject. Item 34. The method of item 30, wherein the TCR recognition domain comprises a peptide autologous to the transplanted cells. Item 35. The method of any one of items 32 or 34, wherein the transplant is a revascularized combined allograft (VCA). Item 36. The method of any of items 35, wherein the autoimmune disease is type 1 diabetes, multiple sclerosis, rheumatoid arthritis, or scleroderma.

[0267] Some aspects of the technology described herein can be defined by the following numbered items: Section 1. a. A TCR recognition domain; b. an intracellular signaling domain, and c. Type 1 biomolecular interaction domains and one or both of A composition comprising: Section 2. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a signaling polypeptide comprising a second type of biomolecular interaction domain and an intracellular signaling domain; A composition comprising: the first type and the second type biomolecular interaction domains specifically bind to each other; composition. Section 3. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a recognition polypeptide comprising a second recognition domain and a third type of biomolecular interaction domain; A composition comprising: the first type and the third type biomolecule interaction domains specifically bind to each other; composition. Section 4. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a signaling polypeptide comprising a second type of biomolecular interaction domain and an intracellular signaling domain; c. a recognition polypeptide comprising a second recognition domain and a third type of biomolecular interaction domain; A composition comprising: the second type and the third type biomolecular interaction domains compete for binding to the first type biomolecular interaction domain; composition. Item 5. The composition according to any one of items 3 to 4, wherein the third type biomolecular interaction domain and the first type biomolecular interaction domain have a higher affinity for each other than the second type biomolecular interaction domain and the first type biomolecular interaction domain. Section 6. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a signaling polypeptide comprising a second type of biomolecular interaction domain, a fourth type of biomolecular interaction domain, and an intracellular signaling domain; c. a recognition polypeptide comprising a second recognition domain and a fifth type biomolecular interaction domain; A composition comprising: the first type biomolecule interaction domain and the second type biomolecule interaction domain specifically bind to each other; the fourth type biomolecule interaction domain and the fifth type biomolecule interaction domain specifically bind to each other; composition. Item 7. The composition of item 6, wherein the fourth type of biomolecular interaction domain and the fifth type of biomolecular interaction domain have weaker affinity than the second type of biomolecular interaction domain and the first type of protein interaction domain. Item 8. The composition according to any one of items 6 to 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, and they specifically bind to each other. Item 9. The composition of any one of items 2 to 8, wherein the first polypeptide comprises the entire TCR recognition domain. Paragraph 10. The composition of any one of paragraphs 2 to 8, wherein the TCR recognition domain comprises at least two distinct polypeptide sequences, the first polypeptide comprises at least one of the distinct polypeptide sequences of the TCR recognition domain, and the first polypeptide binds to or complexes with a second or further polypeptide sequence of the TCR recognition domain to form the TCR recognition domain. Item 11. The composition of any one of items 1 to 10, wherein the TCR recognition domain comprises a non-polypeptide component. Item 12. The composition of any one of items 3 to 11, wherein the second recognition domain is specific for a target not recognized by the TCR recognition domain. Item 13. The composition of any one of items 3 to 12, wherein the second recognition domain is specific for a target found on healthy cells and / or non-target cells and not found on diseased cells and / or target cells. Item 14. The composition of any one of the preceding items, wherein the TCR recognition domain comprises an MHC (major histocompatibility complex), an MHC-peptide complex, a featureless peptide MHC, or an MHC-peptide fusion. Item 15. The composition of item 14, wherein the peptide is a human peptide or a non-human peptide. Item 16. The composition according to any one of items 14 to 15, wherein the peptide is a minor histocompatibility antigen (MiHA). Item 17. The composition of any one of items 14 to 16, wherein the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. Item 18. The composition of any one of items 14 to 17, wherein the MHC-peptide complex is in a monomeric, dimeric, trimeric, tetrameric, pentameric, dextramular, or other oligomeric form. Paragraph 19. The composition of any one of paragraphs 14 to 17, wherein the MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. Item 20. The composition of any one of items 14 to 17, wherein the MHC is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. Item 21. The composition of any one of items 14 to 17, wherein the MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. Item 22. The composition of any one of items 14 to 17, wherein the MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. Item 23. The composition according to any one of items 14 to 22, wherein the MHC is MHC class I or MHC class II. Item 24. The composition of any one of items 1 to 13, wherein the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion. Item 25. The composition described in item 24, wherein said CD1 is CD1d. Item 26. The composition of any one of the preceding items, wherein the biomolecular interaction domain is found in the extracellular portion of the respective polypeptide. Item 27. a. The biomolecular interaction domain is a leucine zipper or any biomolecular interaction domain binding pair is a leucine zipper pair as a whole; b. the biomolecular interaction domain is BZip(RR) and / or AZip(EE), or either biomolecular interaction domain binding pair is BZip(RR) and AZip(EE) as a whole; c. the biomolecular interaction domain is a PSD95-Dlg1-zo-1 (PDZ) domain; d. the biomolecular interaction domain is streptavidin and / or a streptavidin-binding biomolecule (SBP), or either biomolecular interaction domain binding pair is collectively streptavidin and a streptavidin-binding biomolecule (SBP); e. the biomolecular interaction domain is the FKBP binding domain (FRB) of mTOR and / or the FK506 binding biomolecule (FKBP), or any biomolecular interaction domain binding pair is the FKBP binding domain (FRB) of mTOR and the FK506 binding biomolecule (FKBP) as a whole; f. the biomolecular interaction domain is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and / or an FK506-binding biomolecule (FKBP), or either biomolecular interaction domain binding pair is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and an FK506-binding biomolecule (FKBP) as a whole; g. the biomolecular interaction domain is calcineurin A (CNA) and / or FK506-binding biomolecule (FKBP), or the binding pair of either biomolecular interaction domain is calcineurin A (CNA) and FK506-binding biomolecule (FKBP) as a whole; h. The biomolecular interaction domain is gibberellin insensitive (GIA) and / or gibberellin insensitive dwarf1 (GID1), or the binding pair of either biomolecular interaction domain is gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) as a whole; i. the biomolecular interaction domain is a Snap tag and / or a Halo tag, or either biomolecular interaction domain binding pair is collectively a Snap tag and a Halo tag; j. the biomolecular interaction domain is T14-3-3-cΔC and / or the C-terminal peptide of PMA2 (CT52), or the entire binding pair of either biomolecular interaction domain is T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52); k. the biomolecular interaction domain is a PYL and / or an ABI, or any biomolecular interaction domain binding pair is a PYL and an ABI as a whole; l. the biomolecular interaction domain is a nucleotide tag and / or a zinc finger domain, or either biomolecular interaction domain binding pair is collectively a nucleotide tag and a zinc finger domain; m. the biomolecular interaction domain is a nucleotide tag or any biomolecular interaction domain binding pair is collectively a pair of nucleotide tags; n. the biomolecule interaction domain is fluorescein isothiocyanate (FITC) and / or a FITC-binding biomolecule, or any binding pair of the protein interaction domain is FITC and a FITC-binding protein as a whole; and / or o. the protein interaction domain is (R)-phycoerythrin (R-PE / PE) and / or an R-PE / PE binding protein, or any binding pair of the protein interaction domain is (R)-phycoerythrin (R-PE / PE) and an R-PE / PE binding protein as a whole; The composition according to any one of the preceding items. Item 28. The composition of item 27, wherein the nucleotide tag is a DNA tag or a dsDNA tag. 29. The intracellular signaling domain comprises: TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54(ICAM); CD83; 0);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 10. The composition of any preceding claim, comprising or being a signaling domain from one or more proteins selected from the group consisting of: Item 30. A cell comprising and / or expressing the composition of any one of the preceding items. Item 31. A composition comprising a first polypeptide of any one of the preceding items and a cell that expresses or contains a signaling polypeptide of any one of the preceding items. Paragraph 32. The cell or composition of any one of paragraphs 30-31, wherein the TCR recognition domain comprises an MHC that is allogeneic, autologous, or xenogeneic to the cell. Paragraph 33. The cell or composition of any one of paragraphs 30 to 32, wherein the TCR recognition domain comprises a synthetic MHC. Paragraph 34. The cell or composition of any one of paragraphs 30 to 33, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is allogeneic, autologous, or xenogeneic to the cell. Item 35. The cell or composition of any one of items 30 to 34, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is a synthetic peptide. Item 36. The cell or composition of any one of items 30 to 35, wherein the cell is an NK cell, a dendritic cell, a regulatory T cell, or an effector T cell. Paragraph 37. The cell or composition of any one of paragraphs 30 to 36, wherein the cell has been modified to express one or more of the polypeptides of the composition. Paragraph 38. The cell or composition of any one of paragraphs 30 to 37, wherein the cell has been modified to express the signaling polypeptide of the composition. Paragraph 39. The cell or composition of any one of paragraphs 30 to 38, wherein the cell has been further modified to knock out or knock down native MHC I / II. Paragraph 40. The cell or composition of any one of paragraphs 30 to 39, wherein the cell has been further modified to knock down native MHC I / II expressed on the cell surface. Section 41. a. Anti-CD127 and / or anti-CD45RO recognition domains; b. Intracellular signaling domain A chimeric antigen receptor (CAR) comprising: Section 42. a. anti-CD127 and / or anti-CD45RO recognition domains, and b. Type 1 protein interaction domains a first polypeptide comprising: c. a second type of protein interaction domain, and d. Intracellular signaling domain and a second polypeptide comprising A composition comprising: the first type protein interaction domain and the second type protein interaction domain specifically bind to each other; composition. Section 43. a. an anti-CD127 recognition domain, and b. Type 1 protein interaction domains a first polypeptide comprising: c. an anti-CD45RO recognition domain, and d. Type 5 protein interaction domains and a second polypeptide comprising: e. Type 2 and Type 4 protein interaction domains, and f. Intracellular signaling domain and a third polypeptide comprising A composition comprising: the first type protein interaction domain and the second type protein interaction domain specifically bind to each other; the fourth type protein interaction domain and the fifth type protein interaction domain specifically bind to each other; composition. Paragraph 44. A cell comprising the CAR or composition described in any one of paragraphs 41 to 43. Section 45. A method of treating or preventing an autoimmune disease or condition, a T cell-mediated inflammatory or immune response, a malignant T cell condition, a transplant rejection, or GvHD in a subject in need thereof, comprising administering to the subject a composition and / or cells described in any one of the preceding sections. 46. ​​The method of claim 45, wherein the TCR recognition domain comprises an MHC allogeneic to the subject. 47. The method of claim 45, wherein the TCR recognition domain comprises an MHC autologous to the transplanted cells. 48. The method of claim 45, wherein the TCR recognition domain comprises an MHC that is heterologous to the transplanted cells. 49. The method of claim 45, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is allogeneic to the subject. 50. The method of claim 45, wherein the TCR recognition domain comprises an MHC and a peptide, the peptide being autologous to the transplanted cells. 51. The method of claim 45, wherein the TCR recognition domain comprises an MHC and a peptide, the peptide being autologous to the transplanted cells. Paragraph 52. The method of any one of paragraphs 45 to 51, wherein the MHC and / or the peptide is synthetic. Item 53. The method of any one of items 45 to 52, wherein the graft is any human or non-human cell, tissue, or organ. Item 54. The method of any one of items 45 to 53, wherein the transplant is an allogeneic hematopoietic stem cell or solid organ transplant. Paragraph 55. The method of any one of paragraphs 45 to 52, wherein the malignant T-cell condition is T-cell acute lymphoblastic leukemia or T-cell lymphoblastic lymphoma. Item 56. The method of any one of items 45 to 52, wherein the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma. Item 57. The autoimmune disease is thyroiditis, type 1 diabetes, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barré syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis, e.g., acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immune-mediated arthritis, chronic inflammatory arthritis, osteoarthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post-treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, spondyloarthritis, and juvenile-onset rheumatoid ... and juvenile-onset rheumatoid arthritis. and ankylosing spondylitis), relapsing arthritis, inflammatory hyperproliferative skin diseases, psoriasis, such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and nail psoriasis, atopy including atopic diseases, such as hay fever and Job's syndrome, dermatitis, such as contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, nonspecific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic endophthalmitis Inflammatory diseases, urticaria, e.g., chronic allergic urticaria and chronic idiopathic urticaria, e.g., chronic autoimmune urticaria, myositis, polymyositis / dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic sclerosis), sclerosis, e.g., systemic sclerosis, multiple sclerosis (MS), e.g., optic-spinal MS, primary progressive MS (PPMS), and relapsing-remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, disseminated sclerosis, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (e.g., Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis (e.g., ulcerative colitis, ulcerative colitis, microscopic colitis, collagenous colitis, polypoid colitis, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), enteritis, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome (e.g., adult or acute respiratory distress syndrome (ARDS)), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, autoimmune blood disorders, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as seen in meningitis, herpes gestationis, pemphigoid of gestationis, scrotal pruritus,autoimmune premature ovarian failure, sudden hearing loss due to autoimmune conditions, 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, non-granulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome, such as chronic or acute glomerulonephritis, e.g. primary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membranoproliferative or membranoproliferative GN (MPGN) including types I and II, and rapidly progressive GN, proliferative nephritis, autoimmune polyendocrine deficiency, balanitis such as plasma cell circumscribed balanitis, balanoposthitis, erythema annulare centrifugally, erythema dyschromatosis perstans, erythema multiforme, granuloma annulare, lichen sclerosus and atrophicus, lichen simplex chronicus, lichen spinous, lichen planus, ichthyosis lamellar, epidermolytic keratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, eczema, allergic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and bullous palmoplantar eczema; asthma, e.g., bronchial asthma, bronchial asthma, and autoimmune asthma; conditions involving T cell infiltration and chronic inflammatory responses; immune responses to foreign antigens such as fetal ABO blood group during pregnancy; chronic pulmonary inflammatory diseases; autoimmune myocarditis; leukocyte adhesion deficiency; lupus, e.g., lupus nephritis, lupus encephalitis, childhood lupus, non-renal lupus, extrarenal lupus, discoid lupus and discoid lupus erythematosus; lupus alopecia; systemic lupus erythematosus (SL) E), such as cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and disseminated lupus erythematosus, juvenile-onset (type I) diabetes, such as insulin-dependent diabetes mellitus (IDDM) in children, adult-onset diabetes (type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic aortopathy, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, sarcoidosis, granulomatous diseases, such as lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitis, such as vasculitis,Necrotizing vasculitis such as large-vessel vasculitis (including polymyalgia rheumatica and giant cell (Takayasu) arteritis), medium-sized vasculitis (including Kawasaki disease and polyarteritis nodosa / periarteritis nodosa), microscopic polyarteritis, immune-mediated vasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, systemic necrotizing vasculitis, and ANCA-associated vasculitis such as Churg-Strauss vasculitis or Churg-Strauss syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs-positive anemia, Diamond-Blackfan anemia, hemolytic anemia, or immune hemolytic anemias such as autoimmune hemolytic anemia (AIHA), pernicious anemia (pernicious anemia), Addison's disease, pure red cell anemia or pure red cell aplasia (PRCA), factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases with leukocyte leakage, CNS inflammatory disorders, multiple organ injury syndromes such as those secondary to sepsis, trauma or hemorrhage, antigen-antibody complex mediated diseases, antiglomerular basement membrane disease, antiphospholipid syndrome, allergic neuritis, Behcet's disease / syndrome, Castleman syndrome, Goodpasture's syndrome, Raynaud's syndrome, Sjögren's syndrome, Stevens-Johnson syndrome, pemphigoid, e.g., bullous pemphigoid and cutaneous pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, mucous membrane pemphigoid, and pemphigus erythematous), autoimmune polyendocrinopathy, Reiter's disease or Reiter's syndrome, immune complex diseases such as immune complex nephritis, antibody-related nephritis, chronic neuropathy such as polyneuropathy, IgM polyneuropathy or IgM-related neuropathy, and autoimmune or immune-mediated thrombocytopenia, e.g., chronic ITP or idiopathic thrombocytopenic purpura (ITP) including acute ITP, scleritis, e.g. idiopathic keratoscleritis, episcleritis, autoimmune diseases of the testes and ovaries, e.g. autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis, e.g. autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis) or subacute thyroiditis, idiopathic hypothyroidism, Graves' disease, polyglandular syndromes, e.g. autoimmune polyglandular syndrome (or polyendocrinopathy syndrome), paraneoplastic syndromes including neurological paraneoplastic syndromes,Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis, e.g. allergic encephalomyelitis or allergic encephalomyelitis 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 syndrome, autoimmune hepatitis, lupoid hepatitis, giant cell hepatitis, autoimmune chronic active hepatitis, lymphocytic interstitial pneumonia (LIP), bronchiolitis obliterans (non-transplant) vs. NSIP, Guillain-Barré syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis, e.g. primary biliary cirrhosis and pulmonary cirrhosis, autoimmune enteropathy syndrome, celiac disease or celiac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, muscle atrophy Autoimmune lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear diseases such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsing or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome / non-syphilitic interstitial keratitis, Bell's palsy, Sweet's disease / syndrome, autoimmune rosacea, shingles-associated pain, amyloidosis, non-cancerous lymphocytosis, monoclonal antibody tests primary lymphocytosis including central B-cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathies, paraneoplastic syndromes, channelopathies including CNS channelopathies, autism, inflammatory myopathies, focal or segmental glomerulosclerosis or focal segmental glomerulosclerosis (FSGS), endocrine ophthalmopathy, uveitis, chorioretinitis, autoimmune liver disease, fibromyalgia, polyendocrine deficiency, Schmidt syndrome, adrenalitis, gastrotrophy, presenile dementia, autoimmune demyelinating diseases and demyelinating diseases such as chronic inflammatory demyelinating polyneuropathy, Dressler syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility,for example due to antisperm antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortions, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, fancier's lung, allergic granulomatous vasculitis, benign lymphocytic vasculitis, Alport's syndrome, alveolitis, e.g. allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reactions, Samter's syndrome, Kaplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial pulmonary fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum elevata, erythroblastosis fetalis, eosinophilic fasciitis, Charmant's syndrome group, Felty's syndrome, cyclitis, e.g., chronic cyclitis, heterochromic iritis, iridocyclitis (acute or chronic), or Fuchs' cyclitis, Henoch-Schönlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndrome, Evans' syndrome, autoimmune dysgonadism, Sydenham's chorea, post-streptococcal nephritis, thromboangiitis obliterans, thyrotoxicosis, spinal cord fistula, choroiditis, giant cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplanted organ reperfusion, autoimmune retinopathy, aphthous ulcers, Aphthous stomatitis, arteriosclerotic disorders, azoospermia, autoimmune hemolysis, Beck's disease, allergic enterocolitis, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, rheumatic fever, Hammann-Rich disease, sensorineural hearing loss, focal ileitis, leukopenia, transverse myelitis, primary idiopathic myxedema, sympathetic ophthalmia, acute polyradiculitis, pyoderma gangrenosum, acquired splenic atrophy, vitiligo, toxic shock syndrome, conditions involving T-cell infiltration, leukocyte adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, diseases involving leukocyte leakage, multiple organ injury syndrome, Antigen-antibody complex mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathy, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine deficiency, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid sinusitis, frontal sinusitis, maxillary sinusitis or sphenoid sinusitis, eosinophil-related disorders such as eosinophilia, eosinophilic pulmonary infiltrates, eosinophilic myalgia syndrome, Löffler's syndrome,The method according to any one of items 45 to 52, wherein the disease is chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, eosinophil-containing granuloma, seronegative spondyloarthritis, autoimmune polyglandular disease, sclerosing cholangitis, sclera, episclera, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia-telangiectasia syndrome, vascular ectasia, autoimmune disorders associated with connective tissue diseases, rheumatism, allergic hypersensitivity disorders, glomerulonephritis, reperfusion injury, ischemia-reperfusion injury, lymphomatous tracheobronchitis, inflammatory skin diseases, skin diseases with an acute inflammatory component, and autoimmune uveoretinitis (AUR). Item 58. The method of any one of items 45 to 52, wherein the T cell-mediated immune response is an anti-drug specific response to a biologic, cell therapy, and / or gene therapy. Item 59. The method of item 58, wherein the biologic, cell therapy, or gene therapy is adeno-associated virus (AAV) gene therapy, genome editing agent, or enzyme replacement therapy. Clause 60. The method of any one of clauses 45 to 52, wherein the disease is type 1 diabetes and the TCR recognition domain comprises a sequence having 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. Clause 61. The method of any one of clauses 45 to 52, wherein the disease is vitiligo and the TCR recognition domain comprises a sequence having at least 80% or at least 95% sequence identity to SEQ ID NO:18, SEQ ID NO:19, and one of SEQ ID NOs:20-22, or comprises HLA-A*0201 and SEQ ID NO:2018, or HLA-A*0301 and SEQ ID NO:2019, or HLA-A*2402 and SEQ ID NO:2020, or HLA-A*0101 and SEQ ID NO:2021. Clause 62. The method of any one of clauses 45 to 52, wherein the method is for treating and / or preventing GvHD, and the TCR recognition domain comprises a sequence having at least 80% or at least 95% sequence identity to at least one of HLA-A*0101 and SEQ ID NO:2034 to 2037; or HLA-B*0702 and SEQ ID NO:2038; or HLA-B*0801 and SEQ ID NO:2039. Clause 63. The method of any one of clauses 45 to 52, wherein the disease is type 1 diabetes and the TCR recognition domain comprises one or more of SEQ ID NOs:8-17, or 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. Clause 64. The method of any one of clauses 45 to 52, wherein the disease is vitiligo and the TCR recognition domain comprises SEQ ID NO:18, SEQ ID NO:19, and one of SEQ ID NOs:20 to 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. Clause 65. The method of any one of clauses 45 to 52, wherein the method is a method for treating and / or preventing GvHD, and the TCR recognition domain comprises at least one of HLA-A*0101 and SEQ ID NO:2034 to 2037, or HLA-B*0702 and SEQ ID NO:2038, or HLA-B*0801 and SEQ ID NO:2039.

[0268] Some aspects of the technology described herein can be defined by the following numbered items: Section 1. a. A TCR recognition domain; b. an intracellular signaling domain, and c. Biomolecular interaction domains and one or both of A composition comprising: Item 2. The composition described in item 1, comprising the TCR recognition domain and the biomolecular interaction domain. Item 3. The composition described in item 1, comprising the TCR recognition domain and the intracellular signaling domain. 4. 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; the first type and the second type biomolecular interaction domains specifically bind to each other; 2. The composition of claim 1. Item 5. The composition of item 1, wherein the TCR recognition domain comprises an MHC (major histocompatibility complex), an MHC-peptide complex, a featureless peptide-MHC, or an MHC-peptide fusion. Item 6. The composition of item 1, wherein the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion. Item 7. The composition described in item 6, wherein said CD1 is CD1d. Item 8. The composition of item 1, wherein the TCR recognition domain is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form. Item 9. a. The biomolecular interaction domain is a leucine zipper or any biomolecular interaction domain binding pair is a leucine zipper pair as a whole; b. the biomolecular interaction domain is BZip(RR) and / or AZip(EE), or either biomolecular interaction domain binding pair is BZip(RR) and AZip(EE) as a whole; c. the biomolecular interaction domain is a PSD95-Dlg1-zo-1 (PDZ) domain; d. the biomolecular interaction domain is streptavidin and / or a streptavidin-binding biomolecule (SBP), or either biomolecular interaction domain binding pair is collectively streptavidin and a streptavidin-binding biomolecule (SBP); e. the biomolecular interaction domain is the FKBP binding domain (FRB) of mTOR and / or the FK506 binding biomolecule (FKBP), or any biomolecular interaction domain binding pair is the FKBP binding domain (FRB) of mTOR and the FK506 binding biomolecule (FKBP) as a whole; f. the biomolecular interaction domain is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and / or an FK506-binding biomolecule (FKBP), or either biomolecular interaction domain binding pair is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and an FK506-binding biomolecule (FKBP) as a whole; g. the biomolecular interaction domain is calcineurin A (CNA) and / or FK506-binding biomolecule (FKBP), or the binding pair of either biomolecular interaction domain is calcineurin A (CNA) and FK506-binding biomolecule (FKBP) as a whole; h. The biomolecular interaction domain is gibberellin insensitive (GIA) and / or gibberellin insensitive dwarf1 (GID1), or the binding pair of either biomolecular interaction domain is gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) as a whole; i. the biomolecular interaction domain is a Snap tag and / or a Halo tag, or either biomolecular interaction domain binding pair is collectively a Snap tag and a Halo tag; j. the biomolecular interaction domain is T14-3-3-cΔC and / or the C-terminal peptide of PMA2 (CT52), or the entire binding pair of either biomolecular interaction domain is T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52); k. the biomolecular interaction domain is a PYL and / or an ABI, or any biomolecular interaction domain binding pair is a PYL and an ABI as a whole; l. the biomolecular interaction domain is a nucleotide tag and / or a zinc finger domain, or either biomolecular interaction domain binding pair is collectively a nucleotide tag and a zinc finger domain; m. the biomolecular interaction domain is a nucleotide tag or any biomolecular interaction domain binding pair is collectively a pair of nucleotide tags; n. the biomolecule interaction domain is fluorescein isothiocyanate (FITC) and / or a FITC-binding biomolecule, or any binding pair of the protein interaction domain is FITC and a FITC-binding protein as a whole; and / or o. the protein interaction domain is (R)-phycoerythrin (R-PE / PE) and / or an R-PE / PE-binding protein, or any binding pair of the protein interaction domain is (R)-phycoerythrin (R-PE / PE) and an R-PE / PE-binding protein as a whole; 2. The composition of claim 1. Item 10. The composition of item 9, wherein the nucleotide tag is a DNA tag or a dsDNA tag. Item 11. The composition of item 2, further comprising a cell that expresses or contains a signaling polypeptide. Item 12. The composition of item 11, wherein the TCR recognition domain is allogeneic, autologous, or xenogeneic to the cell. Item 13. The composition of item 11, wherein the TCR recognition domain is a synthetic peptide. Item 14. The composition of item 11, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is allogeneic, autologous, or xenogeneic to the cell. Item 15. The composition of item 11, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is a synthetic peptide. Item 16. The composition of item 11, wherein the cell is an NK cell, a dendritic cell, a regulatory T cell, or an effector T cell. Item 17. The composition of item 11, wherein the cells are further modified to knock down native MHC I / II expressed on the cell surface. Section 18. A method for treating or preventing an autoimmune disease or condition, a T cell-mediated inflammatory or immune response, a malignant T cell condition, a transplant rejection, or GvHD in a subject in need thereof, the method comprising administering to the subject the composition and / or cells described in section 1. Item 19. The method of item 18, wherein the transplant is an allogeneic hematopoietic stem cell or solid organ transplant. 20. The method of claim 18, wherein the malignant T-cell condition is T-cell acute lymphoblastic leukemia or T-cell lymphoblastic lymphoma. Item 21. The method of item 18, wherein the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma. Item 22. The method of item 18, wherein the T cell-mediated immune response is an anti-drug specific response to a biologic, cell therapy, and / or gene therapy. Item 23. The method of item 22, wherein the biologic, cell therapy, or gene therapy is adeno-associated virus (AAV) gene therapy, genome editing agent, or enzyme replacement therapy. 24. The method of claim 18, wherein the disease is type 1 diabetes and the TCR recognition domain comprises a sequence having 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 claim 18, wherein the disease is vitiligo and the TCR recognition domain comprises a sequence having at least 95% sequence identity to one of SEQ ID NO:18 and SEQ ID NO:19 and SEQ ID NOs: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. Item 26. The method of item 18, wherein the method is a method for treating and / or preventing GvHD, and the TCR recognition domain comprises a sequence having at least 95% sequence identity to at least one of HLA-A*0101 and SEQ ID NO:2034 to 2037; or HLA-B*0702 and SEQ ID NO:2038; or HLA-B*0801 and SEQ ID NO:2039. [Example]

[0269] Example 1 Autoreactive and alloreactive T cells that attack patient or donor cells or organs are the primary cause of autoimmunity and graft rejection. Current treatments require severe immunosuppressive therapy, which can lead to severe side effects. Specifically, depletion of autoreactive and alloreactive T cells prevents autoimmunity and graft rejection without immunosuppressive modalities. T cells engineered with chimeric antigen ligands (CARs) can redirect their specificity to pathological T cells. T cells engineered with chimeric antigen receptors (CARs) can redirect their specificity and have already been approved to treat some types of B-cell malignancies. Engineered regulatory T cells that can inhibit immune responses in an antigen-dependent manner are currently being explored to expand the application of CAR T cell therapy, for example, to autoimmune diseases and graft rejection.

[0270] Described herein are CALs and / or CARs that can recognize and bind to specific T cell receptors on disease-causing T cells to specifically deplete disease-causing T cells. The CALs and / or CARs are composed of a peptide-HLA (e.g., a monomer or multimer or oligomer thereof) as a recognition domain fused to a signaling domain derived from a T cell receptor. A split version of this system can also be created, in which the system is composed of two pieces. One piece is a universal CAR, with a T cell signaling domain as the intracellular portion and a biomolecular interaction domain as the extracellular portion. The second piece is a CAL composed of an adaptor molecule, e.g., a peptide-HLA oligomer (or monomer or multimer) fused to a cognate biomolecular interaction domain.

[0271] Many CAR T cell therapies for autoimmune diseases and transplant rejection are designed to use regulatory T cells and target organs to achieve organ-specific (e.g., local) immunosuppression. In contrast, the present design (e.g., CAR design) targets the source immune cells in the patient that cause the disease. Thus, the present invention provides a cell-based treatment for autoimmune diseases and transplant rejection.

[0272] Example 2 Dendritic cells (DCs) are an interconnected link between the innate and adaptive immune systems and can induce protective immune responses after stimulation with various stimuli. DCs can be classified into conventional DCs and plasmacytoid DCs (pDCs) based on their phenotype and function. Numerous studies have demonstrated that DCs are essential mediators of proinflammatory or anti-inflammatory (tolerogenic) responses. A subset of DCs that suppress immune responses is commonly known as tolerogenic DCs due to their ability to induce T cell apoptosis, anergy, and regulatory T cells (Tregs).

[0273] A tolerogenic state in DCs (tol-DCs) can be induced using several pharmaceutical 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 possible therapeutic option 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 cardiac allograft rejection in animal models.

[0274] However, the effects of these DCs are not specific: overexpression of inhibitory molecules, including IL-10, TGF-β, CTLA-4, and SOCS1, allows tol-DCs to more efficiently induce Tregs, which may cause systemic immunosuppression.

[0275] Alloreactive immune cells can be specifically suppressed using the methods described herein, for example, by using the universal UniCAR DC system that presents donor pMHC tetra / dextramers (e.g., CAL) and binds genetically modified DCs (K / O MHC I / II but presenting the donor peptide on the tetra / dextramers). In some embodiments, the pMHC can be provided as a monomer, oligomer, or multimer.

[0276] To suppress alloreactive T cells in the recipient, any form of recipient pMHC (e.g., monomer / oligomer / tetramer / dextramers) plus a targeting molecule (e.g., CAL) can be used in conjunction with CAR-DCs. Libraries of identified different alloantigens plus CALs and / or CAR-based MHC monomers / oligomers / tetramers / dextramers associated or associated with targeting molecules (e.g., HLA-A2 plus insulin peptide) can be produced. Such libraries can be commercially available and combined with DCs to target and destroy autoreactive T cells.

[0277] Genetically modified cells can be produced with specific genetic deletions of several genes (IL-12 and NF-κB, MHC I and MHC II [targeting modules would provide these cells with the donor's MHC]) and insertions of several other genes (IL-10, TGF-β, CTLA-4, and SOCS1). These cells are commercially available and can be combined with specific donor pMHC and targeting modules selected from the aforementioned libraries.

[0278] Example 3 Alloreactive memory T cells have been shown to play a key role in activating the recipient immune system and rejecting allogeneic grafts. They are known to be a major barrier to tolerance induction through mixed chimerism and other strategies. These cells are highly resilient, refractory to preconditioning protocols, and rapidly recover from irradiation and T cell depletion. By using CAR-T effectors (conventional, Supra, universal CARs, or CARs) and attaching any form of donor pMHC (monomer, oligomer, tetramer, or dextramer) (e.g., CARs) to the system, the resulting compositions of the present invention can be used to reduce or eliminate the need for improved immunosuppressive therapy in the context of transplantation. Hematopoietic stem cell engraftment can be achieved with durable mixed chimerism with minimal or no toxic preconditioning protocols.

[0279] The use of donor pMHC (monomers / oligomers / tetramers / dextramers, e.g., CALs) in combination with CAR T effectors to suppress allo / xenoreactive T cells would reduce or eliminate the need for immunosuppression. Different donor MHC monomers / oligomers / tetramers (e.g., CALs) attached to the targeting molecules of SUPRA / UNI / universal CALs and / or CAR systems could be produced and commercially available. Shelf-ready libraries of the most common MHC (plus immunological wild-type or synthetic peptide molecules that can be used in combination with that MHC) monomers / oligomers / tetramers could be commercially produced and offered for use with this system.

[0280] Example 4 The involvement of the adaptive immune system in autoimmune diseases has been extensively characterized. T cells are a crucial contributor to autoimmune disease. Conventional T (Teff or T helper) cell subsets, involved in B cell activation and differentiation, produce various inflammatory cytokines and destroy target cells through direct cytotoxicity. CAR-T cells have been used to destroy autoimmune B and T cells in a manner similar to how CD19 CAR T cells target and destroy leukemia cells. Targeting autoreactive memory T and B cells has shown some results. Because these methods are nonspecific, they induce a degree of generalized immunosuppression and are not completely effective. The correlation between specific peptide+MHC molecules and certain autoimmune conditions has been extensively studied. By using available human / animal model pMHC monomers / oligomers / tetramers / dextramers and combining them with SUPRA / UNI / universal CAL and / or CAR technologies, we can target autoreactive immune cells with remarkable specificity.

[0281] The use of some form of autoantigen on pMHC (monomer / oligomer / tetramer / dextramers) (e.g., CALs) in combination with CAR T effectors / Tregs to suppress autoreactive T cells in the recipient would reduce or eliminate the need for immunomodulatory drugs. Libraries of identified autoantigens and related MHC monomers / oligomers / tetramers / dextramers (e.g., HLA-A2+ insulin peptides) attached to targeting molecules on SUPRA / UNI / universal CALs and / or CAR systems can be produced and commercially available for combination with ideal SUPRA or universal CAR T-regs / T effectors to target and destroy autoreactive T cells.

[0282] Example 5 Although new advances have improved survival rates after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) remains a leading cause of long-term morbidity and mortality after transplantation. Current treatment options, especially in steroid-refractory disease, have limited efficacy and lack robust data to inform management decisions.

[0283] Adoptive T cell therapy (ACT) refers to the therapeutic use of T cells. T cells genetically engineered to express chimeric antigen receptors (CARs) represent the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemia and lymphoma, producing remarkable clinical results. The technology described herein offers the opportunity to target disease cells bearing specific antigens or receptors with great precision. In the context of GvHD, universal UNICAR T cells can be designed to find recipient-reactive T cells in donor T cell populations. By fusing the recipient's pMHC to a targeting module that binds to the UniCAR (e.g., thereby forming a CAR), this system can recognize TCR repertoires in donor T / B cell populations that are capable of binding to those MHCs. By fusing the recipient's pMHC to a targeting module that binds to the universal CAR, this system can recognize and bind T cell repertoires in donor T cell populations that are capable of binding to those MHCs.

[0284] If recipient pMHC (monomers / oligomers / tetramers / dextramers) (e.g., CALs) are used in combination with CAR T effectors / Tregs to suppress recipient-reactive T cells in donor HSCs, these reactive T cells will be depleted and GvHD will not occur. Libraries of identified different antigens plus related human / animal model MHC tetramers / dextramers (e.g., HLA-A2+ peptides) attached to targeting molecules of the SUPRA / UNI CAR system can be produced and commercially available for combination with ideal SUPRA CAR T-regs / T effectors to target and destroy recipient-reactive T cells. Libraries of wild-type and / or synthetic MHC monomers / oligomers (e.g., HLA-A2+ peptides) attached to targeting molecules of the universal CAL system can be generated and mixed with universal CAL T-regs / T effectors to target and kill recipient-reactive T cells within the donor T cell population.

[0285] Example 6 Although new advances have improved survival rates after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) remains a leading cause of long-term morbidity and mortality after transplantation. Current treatment options, especially in steroid-refractory disease, have limited efficacy and lack robust data to inform management decisions.

[0286] Adoptive T cell therapy (ACT) refers to the therapeutic use of T cells. T cells genetically engineered to express chimeric antigen receptors (CARs) represent the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemia and lymphoma and have produced remarkable clinical results. This technology offers the opportunity to target cells bearing specific antigens or receptors with great precision. In the context of GvHD, UNICAR T cells can be designed to identify recipient-reactive T cells in donor T cell populations. By fusing the recipient's pMHC to a targeting module that binds to the UniCAR, this system can recognize the TCR repertoire in the donor T / B cell population that can bind to those MHCs.

[0287] If recipient pMHC (tetramers / dextramers) are used in combination with CAR T effectors / Tregs to suppress recipient-reactive T cells in donor HSCs, these cells will be depleted and GvHD will not occur. A library of identified different antigens + related human / animal model MHC tetramers / dextramers (e.g., HLA-A2+ peptides) attached to targeting molecules of the SUPRA / UNI CAR system can be produced and marketed for combination with the ideal SUPRA CAR T-reg / T effector to target and destroy recipient-reactive T cells.

[0288] Example 7 - Modified lymphocytes for preventing pediatric revascularized composite allograft rejection Herein, we describe the development of a clinically applicable tolerance induction regimen for VCA transplantation through the establishment of stable mixed chimerism, augmented by state-of-the-art CAL and / or CAR T cell adoptive immunotherapy and synthetic biology.

[0289] Mixed chimerism in animal models and human allograft recipients has only been shown to be transient, suggesting that tolerance here relies on peripheral inactivation of donor-specific T cells. An obstacle to achieving sustained mixed chimerism and long-term graft acceptance is the presence of high levels of alloreactive memory T cells, which are known to interfere with tolerance induction in sensitized rodents, NHPs, and humans. 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 recipient alloreactive memory immune cells that react with donor bone marrow and tissue cells, thereby aiding in HSCT engraftment and stable mixed chimerism. Selective depletion of memory T lymphocytes by CAL and / or CAR T cell therapy may help achieve sustained mixed chimerism and tolerance, ultimately leading to immunosuppressant-free regimens. Currently, this challenge is addressed by administering a combination of several immunosuppressive drugs in combination with intense total body irradiation, but these result in the depletion of all immune cells as a result of non-specific targeting. This shotgun approach imposes a severe immunocompromised state on the recipient, which subsequently leads to a myriad of consequences, including opportunistic infections and malignancies. The technology herein can specifically delete memory T cells (Aim 1) or only alloreactive memory T cells (Aim 2).

[0290] Aim 1: CD127+ / CD45RO+ memory T cells have important roles as central, effector, and stem cell memory T cells and have been shown to be the most potent component of the alloreactive T cell repertoire. These cells have been shown to be a major contributor to chronic allograft rejection. To address the memory T cell challenge, CAL and / or CAR T-effector cells can be generated using scfv against two common markers of memory T cells (CD127+ and CD45RO+). This will be followed by a mixed chimerism induction protocol in a double-humanized mouse model. Results from this aim will demonstrate that depletion of memory T cells increases the efficacy of mixed chimerism protocols by improving hematopoietic stem cell engraftment, thereby helping to eliminate or reduce the need for immunosuppression in mixed chimerism induction protocols.

[0291] Aim 2: Given the diversity of antigen-specific T cells in the context of transplantation, we provide CAL and / or CAR systems with the flexibility to simultaneously target and attack different alloreactive T cells. CAL and / or CAR T-eff cells and donor pMHC can target alloreactive T cells bearing TCRs for the donor MHC. We will start with monospecific and bispecific antigen-MHC systems. We will then utilize peptide libraries generated from the donor's allograft peptidome and load them onto commercially available exchangeable peptide-MHC multimers. These pMHC and CAL and / or CAR T cell combinations will be used to target and destroy the recipient's alloreactive T cells. This development will result in flexible CAL and / or CAR designs that can target antigen-specific alloreactive T cells.

[0292] Described herein is the investigation of a novel adaptive immune cell therapy strategy for VCA tolerance induction utilizing CAL and / or CAR T cell technology.

[0293] Example 8 - Modified lymphocytes for preventing pediatric revascularized composite allograft rejection Significance: There are nearly 2 million people with limb defects in the United States, and 185,000 amputations occur each year, the majority of which are the result of burns, traffic accidents, and medical conditions. 1 Between 1990 and 2002 alone, 112,600 children were treated in U.S. emergency departments for amputation. Children are often victims of severe burns resulting in limb loss. 2 Over the past decade, revascularized composite allotransplantation (VCA), the transplantation of limbs and face from deceased donors, has become a good alternative option for the reconstruction of devastating injuries to these specialized tissues. VCA is a unique new treatment option for severe soft tissue defects after burns, achieving both psychosocial and functional recovery. 3 The world's first successful pediatric bilateral hand transplant was performed in 2016 between an unrelated donor and recipient pair. 4 The disadvantage is that acute rejection of the "foreign" VCA occurs in up to 90% of patients, despite the use of potent immunosuppressants. 5 Increasing doses of immunosuppression, which are associated with numerous life-threatening complications, are the current approach to preventing VCA loss. 5 .

[0294] During evolution, our immune system acquired the ability to distinguish self from non-self cells and attack "foreign" entities using cells such as T lymphocytes. The human leukocyte antigen (HLA) system (major histocompatibility complex [MHC] in humans) contains cell surface molecules specialized to present antigenic peptides to T cells. 67These peptides are known as minor histocompatibility antigens (MiHA). MiHA sequences can differ between individuals, and many of these differences can be recognized by T cells, initiating the rejection process. T cells are trained to distinguish self-MHC / peptides from non-self-MHC / pept...

Claims

1. a. a TCR recognition domain; b. an intracellular signaling domain, and c. Type 1 biomolecular interaction domains and one or both of A composition comprising:

2. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a signaling polypeptide comprising a second type of biomolecular interaction domain and an intracellular signaling domain; A composition comprising: the first type and the second type biomolecular interaction domains specifically bind to each other; composition.

3. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a recognition polypeptide comprising a second recognition domain and a third type of biomolecular interaction domain; A composition comprising: the first type and the third type biomolecule interaction domains specifically bind to each other; composition.

4. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a signaling polypeptide comprising a second type of biomolecular interaction domain and an intracellular signaling domain; c. a recognition polypeptide comprising a second recognition domain and a third type of biomolecular interaction domain; A composition comprising: the second type and the third type biomolecular interaction domains compete for binding to the first type biomolecular interaction domain; composition.

5. 5. The composition of claim 3, wherein the third type of biomolecular interaction domain and the first type of biomolecular interaction domain have a higher affinity for each other than the second type of biomolecular interaction domain and the first type of biomolecular interaction domain.

6. a. a first polypeptide comprising at least a portion of a TCR recognition domain and a first type of biomolecular interaction domain; b. a signaling polypeptide comprising a second type of biomolecular interaction domain, a fourth type of biomolecular interaction domain, and an intracellular signaling domain; c. a recognition polypeptide comprising a second recognition domain and a fifth type biomolecular interaction domain; A composition comprising: the first type biomolecule interaction domain and the second type biomolecule interaction domain specifically bind to each other; the fourth type biomolecule interaction domain and the fifth type biomolecule interaction domain specifically bind to each other; composition.

7. 7. The composition of claim 6, wherein the fourth type biomolecular interaction domain and the fifth type biomolecular interaction domain have weaker affinity than the second type biomolecular interaction domain and the first type protein interaction domain.

8. The composition of any one of claims 6 to 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, and they specifically bind to each other.

9. The composition of any one of claims 2 to 8, wherein the first polypeptide comprises the entire TCR recognition domain.

10. 9. The composition of any one of claims 2 to 8, wherein the TCR recognition domain comprises at least two distinct polypeptide sequences, the first polypeptide comprises at least one of the distinct polypeptide sequences of the TCR recognition domain, and the first polypeptide binds to or complexes with a second or further polypeptide sequence of the TCR recognition domain to form the TCR recognition domain.

11. The composition of any one of claims 1 to 10, wherein the TCR recognition domain comprises a non-polypeptide component.

12. The composition of any one of claims 3 to 11, wherein the second recognition domain is specific for a target not recognized by the TCR recognition domain.

13. 13. The composition of any one of claims 3 to 12, wherein the second recognition domain is specific for a target found on healthy cells and / or non-target cells and not on diseased cells and / or target cells.

14. 10. The composition of claim 1, wherein the TCR recognition domain comprises an MHC (major histocompatibility complex), an MHC-peptide complex, a featureless peptide MHC, or an MHC-peptide fusion.

15. 15. The composition of claim 14, wherein the peptide is a human peptide or a non-human peptide.

16. The composition of any one of claims 14 to 15, wherein the peptide is a minor histocompatibility antigen (MiHA).

17. 17. The composition of any one of claims 14 to 16, wherein the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form.

18. 18. The composition of any one of claims 14 to 17, wherein the MHC-peptide complex is in a monomeric, dimeric, trimeric, tetrameric, pentameric, dextramular, or other oligomeric form.

19. 18. The composition of any one of claims 14 to 17, wherein the MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form.

20. 18. The composition of any one of claims 14 to 17, wherein the MHC is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form.

21. 18. The composition of any one of claims 14 to 17, wherein the MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form.

22. 18. The composition of any one of claims 14 to 17, wherein the MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer, or other oligomeric form.

23. The composition of any one of claims 14 to 22, wherein the MHC is MHC class I or MHC class II.

24. The composition of any one of claims 1 to 13, wherein the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion.

25. The composition of claim 24, wherein the CD1 is CD1d.

26. 10. The composition of claim 9, wherein the biomolecular interaction domain is found in the extracellular portion of the respective polypeptide.

27. a. the biomolecular interaction domain is a leucine zipper or any biomolecular interaction domain binding pair is a leucine zipper pair as a whole; b. the biomolecular interaction domain is BZip(RR) and / or AZip(EE), or any biomolecular interaction domain binding pair is BZip(RR) and AZip(EE) as a whole; c. the biomolecular interaction domain is a PSD95-Dlg1-zo-1 (PDZ) domain; d. the biomolecular interaction domain is streptavidin and / or a streptavidin-binding biomolecule (SBP), or the binding pair of either biomolecular interaction domain is, as a whole, streptavidin and a streptavidin-binding biomolecule (SBP); e. the biomolecular interaction domain is the FKBP binding domain (FRB) of mTOR and / or the FK506 binding biomolecule (FKBP), or any biomolecular interaction domain binding pair is the FKBP binding domain (FRB) of mTOR and the FK506 binding biomolecule (FKBP) as a whole; f. the biomolecular interaction domain is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and / or an FK506-binding biomolecule (FKBP), or either biomolecular interaction domain binding pair is a cyclophilin-Fas fusion biomolecule (CyP-Fas) and an FK506-binding biomolecule (FKBP) as a whole; g. the biomolecular interaction domain is calcineurin A (CNA) and / or FK506-binding biomolecule (FKBP), or the entire binding pair of any biomolecular interaction domain is calcineurin A (CNA) and FK506-binding biomolecule (FKBP); h. The biomolecular interaction domain is gibberellin insensitive (GIA) and / or gibberellin insensitive dwarf1 (GID1), or the binding pair of any biomolecular interaction domain is gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) as a whole; i. the biomolecular interaction domain is a Snap tag and / or a Halo tag, or either biomolecular interaction domain binding pair is collectively a Snap tag and a Halo tag; j. the biomolecular interaction domain is T14-3-3-cΔC and / or the C-terminal peptide of PMA2 (CT52), or the entire binding pair of either biomolecular interaction domain is T14-3-3-cΔC and the C-terminal peptide of PMA2 (CT52); k. the biomolecular interaction domain is a PYL and / or an ABI, or any biomolecular interaction domain binding pair is a PYL and an ABI as a whole; l. the biomolecular interaction domain is a nucleotide tag and / or a zinc finger domain, or either biomolecular interaction domain binding pair is collectively a nucleotide tag and a zinc finger domain; m. the biomolecular interaction domain is a nucleotide tag or any biomolecular interaction domain binding pair is collectively a pair of nucleotide tags; n. the biomolecular interaction domain is fluorescein isothiocyanate (FITC) and / or a FITC-binding biomolecule, or any binding pair of the protein interaction domain is FITC and a FITC-binding protein as a whole; and / or o. the protein interaction domain is (R)-phycoerythrin (R-PE / PE) and / or an R-PE / PE binding protein, or any binding pair of the protein interaction domain as a whole is (R)-phycoerythrin (R-PE / PE) and an R-PE / PE binding protein; A composition according to any one of the preceding claims.

28. 28. The composition of claim 27, wherein the nucleotide tag is a DNA tag or a dsDNA tag.

29. the intracellular signaling domain is TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; 0);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 10. The composition of any one of the preceding claims, comprising or being a signaling domain from one or more proteins selected from the group consisting of:

30. A cell comprising and / or expressing a composition according to any one of the preceding claims.

31. A composition comprising a first polypeptide according to any one of the preceding claims and a cell that expresses or contains a signaling polypeptide according to any one of the preceding claims.

32. The cell or composition of any one of claims 30 to 31, wherein the TCR recognition domain comprises an MHC that is allogeneic, autologous, or xenogeneic to the cell.

33. The cell or composition of any one of claims 30 to 32, wherein the TCR recognition domain comprises a synthetic MHC.

34. The cell or composition of any one of claims 30 to 33, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is allogeneic, autologous, or xenogeneic to the cell.

35. The cell or composition of any one of claims 30 to 34, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is a synthetic peptide.

36. The cell or composition of any one of claims 30 to 35, wherein the cell is an NK cell, a dendritic cell, a regulatory T cell or an effector T cell.

37. 37. The cell or composition of any one of claims 30 to 36, wherein the cell has been modified to express one or more of the polypeptides of the composition.

38. 38. The cell or composition of any one of claims 30 to 37, wherein the cell has been modified to express the signaling polypeptide of the composition.

39. 39. The cell or composition of any one of claims 30 to 38, wherein the cell has been further modified to knock out or knock down native MHC I / II.

40. 40. The cell or composition of any one of claims 30 to 39, wherein the cell has been further modified to knock down native MHC I / II expressed on the cell surface.

41. a. anti-CD127 and / or anti-CD45RO recognition domains; b. Intracellular signaling domain chimeric antigen receptors (CARs).

42. a. anti-CD127 and / or anti-CD45RO recognition domains, and b. Type 1 protein interaction domains a first polypeptide comprising: c. a second type of protein interaction domain, and d. Intracellular signaling domain and a second polypeptide comprising A composition comprising: the first type protein interaction domain and the second type protein interaction domain specifically bind to each other; composition.

43. a. an anti-CD127 recognition domain, and b. Type 1 protein interaction domains a first polypeptide comprising: c. an anti-CD45RO recognition domain, and d. Type 5 protein interaction domains and a second polypeptide comprising: e. Type 2 and Type 4 protein interaction domains, and f. Intracellular signaling domain and a third polypeptide comprising A composition comprising: the first type protein interaction domain and the second type protein interaction domain specifically bind to each other; the fourth type protein interaction domain and the fifth type protein interaction domain specifically bind to each other; composition.

44. A cell comprising the CAR or composition of any one of claims 41 to 43.

45. A method for treating or preventing an autoimmune disease or condition, a T cell-mediated inflammation or immune response, a malignant T cell condition, a transplant rejection, or GvHD in a subject in need thereof, comprising administering to the subject a composition and / or cells according to any one of the preceding claims.

46. The method of claim 45, wherein the TCR recognition domain comprises an MHC that is allogeneic to the subject.

47. The method of claim 45, wherein the TCR recognition domain comprises an MHC autologous to the transplanted cells.

48. The method of claim 45, wherein the TCR recognition domain comprises an MHC that is heterologous to the transplanted cells.

49. 46. ​​The method of claim 45, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is allogeneic to the subject.

50. The method of claim 45, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is autologous to the transplanted cells.

51. The method of claim 45, wherein the TCR recognition domain comprises an MHC and a peptide, and the peptide is autologous to the transplanted cells.

52. 52. The method of any one of claims 45 to 51, wherein said MHC and / or said peptide is synthetic.

53. The method of any one of claims 45 to 52, wherein the graft is any human or non-human cell, tissue, or organ.

54. The method of any one of claims 45 to 53, wherein the transplant is an allogeneic hematopoietic stem cell or solid organ transplant.

55. 53. The method of any one of claims 45 to 52, wherein said malignant T-cell condition is T-cell acute lymphoblastic leukemia or T-cell lymphoblastic lymphoma.

56. 53. The method of any one of claims 45-52, wherein the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma.

57. The autoimmune disease is selected from the group consisting of thyroiditis, type 1 diabetes, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barré syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis, e.g., acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immune-mediated arthritis, chronic inflammatory arthritis, osteoarthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post-treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, spondyloarthritis, and juvenile-onset rheumatoid arthritis, chronic progressive arthritis, and juvenile-onset rheumatoid arthritis. and ankylosing spondylitis), relapsing arthritis, inflammatory hyperproliferative skin diseases, psoriasis, such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and nail psoriasis, atopy including atopic diseases, such as hay fever and Job's syndrome, dermatitis, such as contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, nonspecific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic intraocular inflammatory disease, Urticaria, e.g. chronic allergic urticaria and chronic idiopathic urticaria, e.g. chronic autoimmune urticaria, myositis, polymyositis / dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic sclerosis), sclerosis, e.g. systemic sclerosis, multiple sclerosis (MS), e.g. optic-spinal MS, primary progressive MS (PPMS), and relapsing-remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, disseminated sclerosis, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (e.g. Crohn's disease, autoimmune-mediated gastrointestinal diseases, colonic ulcers, inflammation, e.g., ulcerative colitis, ulcerative colitis, microscopic colitis, collagenous colitis, polypoid colitis, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), enteritis, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome, e.g., adult or acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, autoimmune blood disorders, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as seen in meningitis, herpes gestationis, pemphigoid of pregnancy, scrotal pruritus, autoimmune premature ovarian failure,sudden hearing loss due to autoimmune conditions, 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, non-granulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome, such as chronic or acute glomerulonephritis, e.g., primary GN, immune-mediated GN, membrane nephritis, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membranoproliferative or membranoproliferative GN (MPGN) including types I and II, and rapidly progressive GN, proliferative nephritis, autoimmune polyendocrine deficiency, balanitis e.g. plasma cell circumscribed balanitis, balanoposthitis, erythema annulare centrifugally, erythema dyschromatosis perstans, erythema multiforme, granuloma annulare, lichen sclerosus and atrophicus, lichen simplex chronicus, lichen spinous, lichen planus, ichthyosis lamellar, epidermolytic keratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, allergic reactions, eczema e.g. allergens asthenic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and bullous palmoplantar eczema; asthma, e.g., bronchial asthma, bronchial asthma, and autoimmune asthma; conditions involving T cell infiltration and chronic inflammatory responses; immune responses to foreign antigens such as fetal ABO blood group during pregnancy; chronic pulmonary inflammatory diseases; autoimmune myocarditis; leukocyte adhesion deficiency; lupus, e.g., lupus nephritis, lupus encephalitis, childhood lupus, non-renal lupus, extrarenal lupus, discoid lupus and discoid lupus erythematosus; lupus alopecia; systemic lupus erythematosus (SLE); for example cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and disseminated lupus erythematosus, juvenile-onset (type I) diabetes, for example insulin-dependent diabetes mellitus (IDDM) of childhood, adult-onset diabetes mellitus (type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic aortopathy, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, sarcoidosis, granulomatous diseases, for example lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitis, for example vasculitis,Necrotizing vasculitis such as large-vasculitis (including polymyalgia rheumatica and giant cell (Takayasu) arteritis), medium-sized vasculitis (including Kawasaki disease and polyarteritis nodosa / periarteritis nodosa), microscopic polyarteritis, immune-mediated vasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, systemic necrotizing vasculitis, and ANCA-associated vasculitis such as Churg-Strauss vasculitis or Churg-Strauss syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs-positive anemia, Diamond-Blackfan anemia, hemolytic anemia, or Immune hemolytic anemias, e.g. autoimmune hemolytic anemia (AIHA), pernicious anemia (pernicious anemia), Addison's disease, pure red cell anemia or pure red cell aplasia (PRCA), factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases with leukocyte leakage, CNS inflammatory disorders, multiple organ injury syndromes, e.g. secondary to sepsis, trauma or hemorrhage, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, antiphospholipid syndrome, allergic neuritis, Behcet's disease / syndrome, Castleman syndrome, Goodpasture's syndrome, Raynaud's syndrome, Sjögren's syndrome, Stevens-Johnson syndrome, pemphigoid, e.g., bullous pemphigoid and cutaneous pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, mucous membrane pemphigoid, and pemphigus erythematous), autoimmune polyendocrinopathy, Reiter's disease or Reiter's syndrome, immune complex diseases such as immune complex nephritis, antibody-related nephritis, chronic neuropathy such as polyneuropathy, IgM polyneuropathy or IgM-related neuropathy, and autoimmune or immune-mediated thrombocytopenia, e.g., chronic ITP or idiopathic thrombocytopenic purpura (ITP) including acute ITP, scleritis, e.g. idiopathic keratoscleritis, episcleritis, autoimmune diseases of the testes and ovaries, e.g. autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis, e.g. autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis) or subacute thyroiditis, idiopathic hypothyroidism, Graves' disease, polyglandular syndromes, e.g. autoimmune polyglandular syndrome (or polyendocrinopathy syndrome), paraneoplastic syndromes including neurological paraneoplastic syndromes,Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis, e.g. allergic encephalomyelitis or allergic encephalomyelitis 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 syndrome, autoimmune hepatitis, lupoid hepatitis, giant cell hepatitis, autoimmune chronic active hepatitis, lymphocytic interstitial pneumonia (LIP), bronchiolitis obliterans (non-transplant) vs. NSIP, Guillain-Barré syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis, e.g., primary biliary cirrhosis and pulmonary cirrhosis, autoimmune enteropathy syndrome, celiac disease or celiac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, muscle atrophy Autoimmune lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear diseases such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsing or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome / non-syphilitic interstitial keratitis, Bell's palsy, Sweet's disease / syndrome, autoimmune rosacea, shingles-associated pain, amyloidosis, non-cancerous lymphocytosis, monoclonal antibody tests primary lymphocytosis, including central B-cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathies, paraneoplastic syndromes, channelopathies, including channelopathies of the CNS, autism, inflammatory myopathy, focal or segmental glomerulosclerosis or focal segmental glomerulosclerosis (FSGS), endocrine ophthalmopathy, uveitis, chorioretinitis, autoimmune liver disease, fibromyalgia, polyendocrine deficiency, Schmidt syndrome, adrenalitis, gastrotrophy, presenile dementia, autoimmune demyelinating diseases and demyelinating diseases such as chronic inflammatory demyelinating polyneuropathy, Dressler syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility,for example due to antisperm antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortions, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, fancier's lung, allergic granulomatous vasculitis, benign lymphocytic vasculitis, Alport's syndrome, alveolitis, e.g. allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reactions, Samter's syndrome, Kaplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial pulmonary fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum elevata, erythroblastosis fetalis, eosinophilic fasciitis, Charmant's syndrome group, Felty's syndrome, cyclitis, e.g., chronic cyclitis, heterochromic iritis, iridocyclitis (acute or chronic), or Fuchs' cyclitis, Henoch-Schönlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndrome, Evans' syndrome, autoimmune dysgonadism, Sydenham's chorea, poststreptococcal nephritis, thromboangiitis obliterans, thyrotoxicosis, spinal cord fistula, choroiditis, giant cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplanted organ reperfusion, autoimmune retinopathy, aphthous ulcers, Aphthous stomatitis, arteriosclerotic disorders, azoospermia, autoimmune hemolysis, Beck's disease, allergic enterocolitis, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, rheumatic fever, Hammann-Rich disease, sensorineural hearing loss, focal ileitis, leukopenia, transverse myelitis, primary idiopathic myxedema, sympathetic ophthalmia, acute polyradiculitis, pyoderma gangrenosum, acquired splenic atrophy, vitiligo, toxic shock syndrome, conditions involving T-cell infiltration, leukocyte adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, diseases involving leukocyte leakage, multiple organ injury syndrome, Antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathy, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine deficiency, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid sinusitis, frontal sinusitis, maxillary sinusitis or sphenoid sinusitis, eosinophil-related disorders such as eosinophilia, eosinophilic pulmonary infiltrates, eosinophilic myalgia syndrome, Löffler's syndrome,The method of any one of claims 45 to 52, wherein the disease is chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, eosinophil-containing granuloma, seronegative spondyloarthritis, autoimmune polyglandular disease, sclerosing cholangitis, sclera, episclerosis, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia-telangiectasia syndrome, vascular ectasia, autoimmune disorders associated with connective tissue diseases, rheumatism, allergic hypersensitivity disorders, glomerulonephritis, reperfusion injury, ischemia-reperfusion injury, lymphomatous tracheobronchitis, inflammatory skin diseases, skin diseases with an acute inflammatory component, and autoimmune uveoretinitis (AUR).

58. 53. The method of any one of claims 45 to 52, wherein the T cell-mediated immune response is an anti-drug specific response to a biologic, cell therapy, and / or gene therapy.

59. 59. The method of claim 58, wherein the biologic, cell therapy, or gene therapy is adeno-associated virus (AAV) gene therapy, a genome editing agent, or an enzyme replacement therapy.

60. The method of any one of claims 45 to 52, wherein the disease is type 1 diabetes and the TCR recognition domain comprises a sequence having at least 80% or at least 95% sequence identity to one or more of SEQ ID NOs:8 to 17; HLA-A*0201 and at least one of SEQ ID NOs:2013 to 2016 and 2031 to 2033; or HLA-A*02:01 and at least one of SEQ ID NOs:20128 to 2129.

61. The method of any one of claims 45 to 52, wherein the disease is vitiligo and the TCR recognition domain comprises a sequence having at least 80% or at least 95% sequence identity to SEQ ID NO:18, SEQ ID NO:19, or one of SEQ ID NOs:20 to 22, or comprises HLA-A*0201 and SEQ ID NO:2018, or HLA-A*0301 and SEQ ID NO:2019, or HLA-A*2402 and SEQ ID NO:2020, or HLA-A*0101 and SEQ ID NO:2021.

62. The method of any one of claims 45 to 52, wherein the method is for treating and / or preventing GvHD and the TCR recognition domain comprises a sequence having at least 80% or at least 95% sequence identity to at least one of HLA-A*0101 and SEQ ID NO:2034 to 2037; or HLA-B*0702 and SEQ ID NO:2038; or HLA-B*0801 and SEQ ID NO:2039.

63. The method of any one of claims 45 to 52, wherein the disease is type 1 diabetes and the TCR recognition domain comprises one or more of SEQ ID NOs:8 to 17, or comprises HLA-A*0201 and at least one of SEQ ID NOs:2013 to 2016 and 2031 to 2033, or comprises HLA-A*02:01 and at least one of SEQ ID NOs:20128 to 2129.

64. The method of any one of claims 45 to 52, wherein the disease is vitiligo and the TCR recognition domain comprises SEQ ID NO:18, SEQ ID NO:19, and one of SEQ ID NOs:20 to 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 one of claims 45 to 52, wherein the method is for treating and / or preventing GvHD and the TCR recognition domain comprises at least one of HLA-A*0101 and SEQ ID NO:2034 to 2037, or comprises HLA-B*0702 and SEQ ID NO:2038, or comprises HLA-B*0801 and SEQ ID NO:2039.

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