Cellular Therapy for Type 1 Diabetes

Abstract

The present disclosure is directed to isolated cell populations comprising a T cell receptor that targets proinsulin and regulatory T cells that exogenously express the T cell receptor for use in treating type 1 diabetes. In some aspects, provided herein is an autologous T cell (e.g., regulatory T cell) therapy for the treatment of type 1 diabetes (T1D), e.g., the treatment of T1D in patients with the HLA-DRB1*04:01 genetic haplotype. In some embodiments, the therapy is a targeted therapy for T1D patients who have residual / residual beta cell function in the pancreas (e.g., still produce insulin).

Description

[Technical Field]

[0001] (Related Applications) This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 63 / 434,062, filed December 20, 2022, U.S. Provisional Patent Application No. 63 / 459,843, filed April 17, 2023, and U.S. Provisional Patent Application No. 63 / 578,226, filed August 23, 2023, the contents of each of which are incorporated herein by reference in their entirety.

[0002] Electronic Sequence Listing Reference The contents of the electronic sequence listing (A133670005WO00-SEQ-MSB.xml, size: 341,248 bytes, and creation date: December 18, 2023) are incorporated herein by reference in their entirety. [Background technology]

[0003] Type 1 diabetes is a T cell-mediated autoimmune disease that results in islet β cell destruction, hypoinsulinemia, and altered glucose homeostasis. The activity of effector T cells and regulatory T cells, which function together in immune homeostasis, is dysregulated in patients with type 1 diabetes. Clinical studies have shown that modulating T cells influences disease progression in these patients. There is evidence that therapeutic intervention in type 1 diabetes patients who have the genetic human leukocyte antigen (HLA) haplotype DR3-DQ2 (more than half of all patients), who are early in the autoimmune pathogenesis, can prevent the development of symptomatic disease and insulin dependence.

[0004] Approximately 64,000 patients are newly diagnosed with type 1 diabetes each year, and there is currently no cure for type 1 diabetes. Available therapies rely on symptomatic treatment, which often involves immunosuppressive agents that can have severe side effects. In these patients, there are no approved therapies that restore immune tolerance and halt the loss of beta cell function. Regulatory T cells have the potential to treat this disease and other autoimmune diseases because they can selectively target affected cell types and tissues and generate local immune responses through antigen-specific mechanisms. Summary of the Invention [Means for solving the problem]

[0005] In some embodiments, provided herein is a T cell therapy (e.g., Treg cell therapy) that uses TCRs to target the pancreas and draining lymph nodes in patients with Type 1 Diabetes (T1D), thereby limiting ongoing beta cell destruction and preserving residual beta cell mass. This therapy has the additional potential to promote repair of damaged tissue and establish long-lived tissue residency. While suppressing T cell function has been shown to delay the onset of T1D, no therapy exists to halt the autoimmunity that drives T1D and permanently recalibrate the immune response. Described herein in some embodiments is a targeted approach to suppressing islet-associated inflammation, providing patients with significant and sustained clinical benefit. In some embodiments, this pioneering approach for patients with T1D exploits the natural role of T cells in the immune system to treat T1D.

[0006] The present disclosure is based, at least in part, on the development of T cell receptors (TCRs) specific for proinsulin (e.g., complexed with a Major Histocompatibility Complex (MHC) molecule), which can be transduced into T cells (e.g., regulatory T cells) and expressed on the surface of these T cells. T cells expressing the TCRs of the present disclosure effectively activate cells (e.g., conventional T cells) in the presence of proinsulin. Thus, these TCRs provide a mechanism for targeting T cells (e.g., regulatory T cells) and the immune system to cells and tissues that express proinsulin (e.g., for the treatment of type 1 diabetes).

[0007] Some embodiments relate to engineered T cell receptors that specifically bind to the preproinsulin 73-90 peptide complexed with an MHC molecule.

[0008] In some embodiments, the engineered T cell receptor comprises an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of any one of SEQ ID NOs: 3, 33, 53, and 73, and / or the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of any one of SEQ ID NOs: 8, 38, 58, and 78.

[0009] In some embodiments, the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO:3, and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:8.

[0010] In some embodiments, the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 1 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 2, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 6 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 7.

[0011] In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 4, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 9. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 4, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 9. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 4, and the beta chain comprises a beta variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 9.

[0012] In some embodiments, the alpha chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 5, and the beta chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 10. In some embodiments, the alpha chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 5, and the beta chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 10. In some embodiments, the alpha chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 5, and the beta chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 10.

[0013] In some embodiments, the alpha chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 260, and the beta chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 261. In some embodiments, the alpha chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 260, and the beta chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 261. In some embodiments, the alpha chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 260, and the beta chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 261.

[0014] In some embodiments, the engineered T cell receptor comprises the sequence of SEQ ID NO: 310. In some embodiments, the engineered T cell receptor comprises the sequence of SEQ ID NO: 311.

[0015] In some embodiments, the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO:33, and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:38.

[0016] In some embodiments, the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 31 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 32, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 36 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 37.

[0017] In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 34, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 39. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 34, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 39. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 34, and the beta chain comprises a beta variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 39.

[0018] In some embodiments, the alpha chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 35, and the beta chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 40. In some embodiments, the alpha chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 35, and the beta chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 40. In some embodiments, the alpha chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 35, and the beta chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 40.

[0019] In some embodiments, the alpha chain comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 266, and the beta chain comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 267. In some embodiments, the alpha chain comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 266, and the beta chain comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 267. In some embodiments, the alpha chain comprises an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 266, and the beta chain comprises an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 267.

[0020] In some embodiments, the engineered T cell receptor comprises the sequence of SEQ ID NO: 312 or 313.

[0021] In some embodiments, the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO:53, and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:58.

[0022] In some embodiments, the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 51 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 52, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 56 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 57.

[0023] In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 54, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 59. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 54, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 59. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 54, and the beta chain comprises a beta variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 59.

[0024] In some embodiments, the alpha chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 55, and the beta chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 60. In some embodiments, the alpha chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 55, and the beta chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 60. In some embodiments, the alpha chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 55, and the beta chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 60.

[0025] In some embodiments, the alpha chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 270, and the beta chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 271. In some embodiments, the alpha chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 270, and the beta chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 271. In some embodiments, the alpha chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 270, and the beta chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 271.

[0026] In some embodiments, the engineered T cell receptor comprises the sequence of SEQ ID NO: 314 or 315.

[0027] In some embodiments, the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO:73, and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:78.

[0028] In some embodiments, the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 71 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 72, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 76 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 77.

[0029] In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 74, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 79. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 74, and the beta chain comprises a beta variable domain comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 79. In some embodiments, the alpha chain comprises an alpha variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 74, and the beta chain comprises a beta variable domain comprising an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 79.

[0030] In some embodiments, the alpha chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 75, and the beta chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 80. In some embodiments, the alpha chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 75, and the beta chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 80. In some embodiments, the alpha chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 75, and the beta chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 80.

[0031] In some embodiments, the alpha chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 274, and the beta chain comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 275. In some embodiments, the alpha chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 274, and the beta chain comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 275. In some embodiments, the alpha chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 274, and the beta chain comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 275.

[0032] In some embodiments, the engineered T cell receptor comprises the sequence of SEQ ID NO: 316 or 317.

[0033] In some embodiments, the engineered T cell receptor is encoded as a single polypeptide.

[0034] In some embodiments, the engineered T cell receptor comprises a self-cleaving peptide sequence located between the alpha and beta chains.

[0035] In some embodiments, the self-cleaving peptide sequence is a 2A peptide sequence, for example, a P2A, E2A, F2A, or T2A peptide sequence.

[0036] In some embodiments, the preproinsulin 73-90 peptide comprises the amino acid sequence GAGSLQPLALEGSLQKRG (SEQ ID NO: 109).

[0037] In some embodiments, the MHC molecule comprises an HLA-DRB1*04:01 molecule.

[0038] Some embodiments relate to an engineered nucleic acid encoding an engineered T cell receptor described in any one of the preceding paragraphs.

[0039] In some embodiments, the engineered nucleic acid is a viral vector, optionally a lentiviral vector.

[0040] In some embodiments, the engineered nucleic acid comprises an EF-1 alpha promoter or an MND promoter operably linked to a sequence encoding an engineered T cell receptor.

[0041] In some embodiments, the engineered nucleic acid further comprises an enhancer element, optionally an optimized Post-Transcriptional Regulatory Element (oPRE) or a Woodchuck Hepatitis Virus Post-Transcriptional Regulatory Element (WPRE), and further optionally a WPRE-mut6.

[0042] Another embodiment relates to regulatory T cells comprising an engineered T cell receptor that specifically binds to the preproinsulin 73-90 peptide complexed with an MHC molecule.

[0043] In some embodiments, the engineered T cell receptor is an engineered T cell receptor described in any one of the preceding paragraphs.

[0044] In some embodiments, the engineered T cell receptor expresses an engineered nucleic acid described in any one of the preceding paragraphs.

[0045] In some embodiments, the regulatory T cells are phenotypically stable.

[0046] In some embodiments, the regulatory T cells have not been gene edited at the endogenous FOXP3 locus.

[0047] Yet another embodiment relates to a method of treating type 1 diabetes in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising regulatory T cells according to any one of the preceding paragraphs.

[0048] In some embodiments, the regulatory T cells are autologous to the subject.

[0049] In some embodiments, the subject has type 1 diabetes, eg, stage 3 type 1 diabetes.

[0050] In some embodiments, the subject is an HLA-DRB1*04:01 positive subject.

[0051] Some embodiments are directed to methods of treating type 1 diabetes in a subject in need thereof, the method comprising administering to a subject diagnosed with type 1 diabetes a therapeutically effective amount of a population of cells comprising regulatory T cells, wherein (i) the regulatory T cells of the population express an engineered T cell receptor, and the engineered T cell receptor is capable of expressing preproinsulin 73-90 (preproinsulin 73-90 or PPI) complexed with an MHC molecule in the pancreas and / or pancreatic-draining lymph nodes of the subject. 73~90 and (ii) less than 10% of the regulatory T cells in the population are gene edited at the endogenous FOXP3 locus.

[0052] Another aspect relates to a method of treating type 1 diabetes in a subject in need thereof, the method comprising administering to a subject diagnosed with type 1 diabetes a therapeutically effective amount of a population of cells comprising regulatory T cells, wherein the regulatory T cells of the population express any one of the engineered T cell receptors of Table 1, and wherein the regulatory T cells specifically bind to preproinsulin 73-90 complexed with an MHC molecule in the pancreas and / or pancreatic-draining lymph nodes of the subject.

[0053] In some embodiments, the population of cells is autologous to the subject.

[0054] In some embodiments, less than 1% of the regulatory T cells in the population have been gene edited at the endogenous FOXP3 locus, e.g., less than 0.5% or less than 0.1% of the regulatory T cells in the population have been gene edited at the endogenous FOXP3 locus.

[0055] In some embodiments, the subject is an HLA-DRB1*04:01 positive subject.

[0056] In some embodiments, the subject has been diagnosed with stage 3 type 1 diabetes.

[0057] In some embodiments, the method further includes selecting a subject who is HLA-DRB1*04:01 positive and has been diagnosed with type 1 diabetes, e.g., stage 3 type 1 diabetes, and administering to the subject a population of cells comprising regulatory T cells described herein.

[0058] In some embodiments, the regulatory T cells of the population are phenotypically stable regulatory T cells. In some embodiments, at least 80% of the cells of the population are phenotypically stable regulatory T cells. For example, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the cells of the population can be phenotypically stable regulatory T cells.

[0059] In some embodiments, no more than 20% of the cells in the population are conventional CD4 + T cells. For example, 15% or less, 10% or less, or 5% or less of the cells in the population are conventional CD4 + It may be a T cell.

[0060] In some embodiments, 5% or less of the cells in the population are CD8 + T cells. For example, less than 1% of the cells in a population are CD8 + It may be a T cell.

[0061] In some embodiments, 10% or less of the cells in the population are non-regulatory T cells that express an engineered T cell receptor. For example, 5% or less or 1% or less of the cells in the population can be non-regulatory T cells that express an engineered T cell receptor.

[0062] In some embodiments, at least 40% of the cells in the population express an engineered T cell receptor. + CD25 hi / + CD127 - / low Regulatory T cells. For example, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the cells in the population are CD4 T cells that express an engineered T cell receptor. + CD25 hi / + CD127 - / low They may be regulatory T cells.

[0063] In some embodiments, at least 70% of the cells in the population are viable. For example, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of the cells in the population can be viable.

[0064] In some embodiments, the method further comprises, prior to administering the cells, isolating the cells from the subject, sorting the cells to obtain phenotypically stable regulatory T cells, and expanding the phenotypically stable regulatory T cells, thereby producing a population of cells phenotypically stable regulatory T cells.

[0065] Some embodiments relate to methods of preventing beta cell destruction and / or preserving beta cell mass in a subject in need thereof, the method comprising administering to the subject a composition comprising a cell population expressing a TCR provided herein, wherein the TCR specifically binds to preproinsulin 73-90 complexed with an MHC molecule (e.g., an MHC class II antigen) in the pancreas and / or draining lymph nodes of the subject, thereby preventing beta cell destruction and / or preserving beta cell mass in the subject.

[0066] Another aspect relates to a method of repairing damaged pancreatic tissue in a subject in need thereof, comprising administering to the subject a composition comprising a cell population expressing a TCR provided herein, wherein the TCR specifically binds to preproinsulin 73-90 complexed with an MHC molecule in the pancreas and / or draining lymph nodes of the subject, thereby repairing damaged pancreatic tissue in the subject.

[0067] Yet another aspect relates to a method of preventing autoimmunity associated with type 1 diabetes in a subject in need thereof, comprising administering to the subject a composition comprising a population of cells expressing a TCR provided herein, wherein the TCR specifically binds to preproinsulin 73-90 complexed with an MHC molecule in the pancreas and / or draining lymph nodes of the subject, thereby preventing autoimmunity associated with type 1 diabetes in the subject.

[0068] Some embodiments relate to a method of suppressing islet-associated inflammation in a subject in need thereof, comprising administering to the subject a composition comprising the cell population of claim 247, wherein a TCR specifically binds to preproinsulin 73-90 complexed with an MHC molecule in the pancreas and / or draining lymph nodes of the subject, thereby suppressing islet-associated inflammation in the subject.

[0069] In some embodiments, the subject has type 1 diabetes. In some embodiments, the subject has stage 3 type 1 diabetes (e.g., new-onset stage 3 type 1 diabetes).

[0070] In some embodiments, the composition further comprises a pharmaceutically acceptable excipient or cryopreservative.

[0071] In some embodiments, the administering comprises intravenous administration. In some embodiments, the administering comprises one or more injections (e.g., one or more injections of the isolated cell population or pharmaceutical composition). In some embodiments, the administering comprises a single injection (e.g., a single injection of the isolated cell population or pharmaceutical composition). In some embodiments, the administering comprises two or more injections of the isolated cell population or pharmaceutical composition.

[0072] In some embodiments, the subject has the HLA-DRB1 genetic haplotype (expressing Major Histocompatibility Complex, Class II, DR Beta 1 (HLA-DRB1)). In some embodiments, the subject has the HLA-DRB1*04:01 genetic haplotype.

[0073] In some embodiments, the subject has new-onset / adult-onset type 1 diabetes. In some embodiments, the subject has new-onset / adult-onset stage 3 type 1 diabetes.

[0074] In some embodiments, the subject has residual beta cell function.

[0075] Some aspects of the present disclosure provide one or more amino acid sequences encoding a TCR that binds (e.g., specifically binds) a proinsulin peptide (e.g., preproinsulin 73-90) complexed with an MHC molecule. In some embodiments, the MHC molecule is an MHC class II antigen.

[0076] Some aspects of the present disclosure provide TCRs that (a) comprise one or more amino acid sequences set forth in Table 1 (e.g., CDR sequences belonging to any one of TCR-A, TCR-B, TCR-C, TCR-D, TCR-E, TCR-F, TCR-G, TCR-H, TCR-I, TCR-J, TCR-K, TCR-L, TCR-M, TCR-N, TCR-O, TCR-P, TCR-Q, TCR-R, TCR-S, TCR-T, TCR-U, TCR-V, TCR-X, TCR-Y, and TCR-Z), and (b) bind (e.g., specifically bind) to a proinsulin peptide (e.g., preproinsulin 73-90) complexed with an MHC molecule. In some embodiments, the proinsulin peptide is preproinsulin 73-90.

[0077] In some embodiments, the preproinsulin 73-90 peptide comprises the amino acid sequence GAGSLQPLALEGSLQKRG (SEQ ID NO: 109). In some embodiments, the MHC molecule comprises an HLA-DRB1*04:01 molecule. In some embodiments, the MHC molecule comprises an HLA-DRA*0101 molecule.

[0078] In some embodiments, the TCR is encoded as a single polypeptide. The polypeptide encoding the TCR may include, for example, an N-terminal beta domain and a C-terminal alpha domain. In some embodiments, the polypeptide includes a self-cleaving peptide sequence located between the TCR alpha chain and the TCR beta chain. In some embodiments, the self-cleaving peptide sequence is a 2A peptide sequence. The 2A peptide sequence may be, for example, a P2A, E2A, F2A, or T2A peptide sequence. In some embodiments, the 2A peptide sequence is a P2A peptide sequence.

[0079] In some embodiments, the TCR alpha chain comprises a murine constant region. In some embodiments, the TCR beta chain comprises a murine constant region.

[0080] In some embodiments, the TCR comprises one or more amino acid substitutions to cysteine ​​residues in the alpha chain constant region and the beta chain constant region, wherein the cysteine ​​residues form or are capable of forming one or more disulfide bonds. In some embodiments, the TCR alpha chain constant region comprises a T48C amino acid substitution relative to the alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, and the beta chain constant region comprises a S57C amino acid substitution relative to the beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108.

[0081] Some aspects of the present disclosure provide a nucleic acid encoding any one of the TCRs described herein. In some embodiments, the nucleic acid is a vector (e.g., a viral vector or a lentiviral vector).

[0082] In some embodiments, the nucleic acid comprises a promoter operably linked to a coding sequence encoding a TCR. The promoter may be, for example, an EF-1 alpha promoter or an MND promoter. In some embodiments, the nucleic acid further comprises an enhancer element. The enhancer element may be, for example, an optimized post-transcriptional regulatory element (oPRE) or a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). In some embodiments, the enhancer element comprises WPRE-mut6. In some embodiments, the coding sequence is codon-optimized.

[0083] Some aspects of the present disclosure provide engineered cells that include a TCR (e.g., an engineered TCR) that binds to (e.g., specifically binds to) a proinsulin peptide (e.g., preproinsulin 73-90) complexed with an MHC molecule. In some embodiments, the proinsulin peptide is preproinsulin 73-90. In some embodiments, the TCR is any one of the TCRs described herein. In some embodiments, the engineered cell is a T cell, e.g., a regulatory T cell. In some embodiments, the engineered cell is from (e.g., isolated from) a subject, for example, from a subject that has (e.g., has been diagnosed with) or is suspected of having type 1 diabetes.

[0084] Some embodiments of the present disclosure provide a cell population comprising a plurality of engineered cells (e.g., regulatory T cells) comprising any one of the TCRs described herein.

[0085] Some aspects of the present disclosure provide pharmaceutical compositions comprising the cell populations described herein and a pharmaceutically acceptable excipient. Other aspects of the present disclosure provide compositions comprising the cell populations described herein and a cryopreservation agent.

[0086] Some aspects of the present disclosure provide methods comprising administering to a subject a population of cells, pharmaceutical composition, or composition described herein, wherein the subject has type 1 diabetes. In some embodiments, the population of cells, pharmaceutical composition, or composition is administered in an amount effective to alleviate one or more symptoms of type 1 diabetes. In some embodiments, the administering comprises intravenous administration. In some embodiments, the administering comprises one or more injections (e.g., one or more injections of a population of cells, pharmaceutical composition, or composition described herein). In some embodiments, the administering comprises a single injection. In some embodiments, the administering comprises a single injection of a population of cells (e.g., an isolated cell population). In some embodiments, the administering comprises a single injection of a pharmaceutical composition.

[0087] In some embodiments, the TCR specifically binds to preproinsulin 73-90 complexed with an MHC molecule (eg, an MHC class II antigen) in the pancreas and / or draining lymph nodes of a subject.

[0088] In some embodiments, the cell population, pharmaceutical composition, or composition is administered in an amount effective to prevent beta cell destruction and / or preserve beta cell mass in the pancreas of a subject.

[0089] In some embodiments, the cell population, pharmaceutical composition, or composition is administered in an amount effective to repair pancreatic tissue damage in a subject.

[0090] In some embodiments, the cell population, pharmaceutical composition, or composition is administered in an amount effective to prevent autoimmunity associated with type 1 diabetes in a subject.

[0091] In some embodiments, the cell population, pharmaceutical composition, or composition is administered in an amount effective to suppress pancreatic islet-associated inflammation in a subject. [Brief explanation of the drawings]

[0092] [Figure 1A] 1 provides a graph showing the ability of TCRs of the present disclosure to activate TCR-null Jurkat cells in the presence of preproinsulin peptide, as indicated by CD69 expression levels. [Figure 1B] 1 provides a graph showing the ability of TCRs of the present disclosure to activate TCR-null Jurkat cells in the presence of preproinsulin peptide, as indicated by CD69 expression levels. [Figure 2] 1 provides a graph showing activation of TCR-null Jurkat cells by TCRs of the present disclosure as determined by CD69 levels at the indicated concentrations of preproinsulin peptide. [Figure 3] 1 provides a graph showing activation of conventional CD4+ T cells by TCRs of the present disclosure as determined by CD69 levels at the indicated concentrations of preproinsulin peptide. [Figure 4A] 1 provides a graph showing activation of conventional CD4+ T cells by TCRs of the present disclosure as determined by the percentage of TCR+ cells expressing CD69 at the indicated concentrations of preproinsulin peptide. [Figure 4B] 1 provides a graph showing activation of conventional CD4+ T cells by TCRs of the present disclosure as determined by the percentage of TCR+ cells expressing CD69 at the indicated concentrations of preproinsulin peptide. [Figure 5A]

[0023] Figure 1 provides a graph showing activation of conventional CD4+ T cells by TCRs of the present disclosure, as determined by the percentage of TCR+ cells expressing CD69, at the indicated concentrations of preproinsulin peptide, where the TCRs have either unmodified or disulfide-modified constant domain sequences. [Figure 5B]

[0023] Figure 1 provides a graph showing activation of conventional CD4+ T cells by TCRs of the present disclosure, as determined by the percentage of TCR+ cells expressing CD69, at the indicated concentrations of preproinsulin peptide, where the TCRs have either unmodified or disulfide-modified constant domain sequences. [Figure 5C]

[0023] Figure 1 provides a graph showing activation of conventional CD4+ T cells by TCRs of the present disclosure, as determined by the percentage of TCR+ cells expressing CD69, at the indicated concentrations of preproinsulin peptide, where the TCRs have either unmodified or disulfide-modified constant domain sequences. [Figure 6] 1 provides a graph showing activation of regulatory T cells by TCRs of the present disclosure as determined by the percentage of TCR+ cells expressing CD69 at the indicated concentrations of preproinsulin peptide. [Figure 7A] 1 provides graphs showing the suppressive activity of regulatory T cells by TCRs of the present disclosure as determined by IL-10 production and CD69 upregulation at various concentrations of preproinsulin peptide. [Figure 7B] 1 provides graphs showing the suppressive activity of regulatory T cells by TCRs of the present disclosure as determined by IL-10 production and CD69 upregulation at various concentrations of preproinsulin peptide. [Figure 7C] 1 provides graphs showing the suppressive activity of regulatory T cells by TCRs of the present disclosure as determined by IL-10 production and CD69 upregulation at various concentrations of preproinsulin peptide. [Figure 7D] 1 provides graphs showing the suppressive activity of regulatory T cells by TCRs of the present disclosure as determined by IL-10 production and CD69 upregulation at various concentrations of preproinsulin peptide. [Figure 7E] 1 provides graphs showing the suppressive activity of regulatory T cells by TCRs of the present disclosure as determined by IL-10 production and CD69 upregulation at various concentrations of preproinsulin peptide. [Figure 8A] 1 provides a graph showing the suppressive activity of regulatory T cells expressing a TCR of the present disclosure on conventional T cells, as determined by their ability to suppress proliferation of conventional T cells. [Figure 8B] 1 provides a graph showing the suppressive activity of regulatory T cells expressing a TCR of the present disclosure on conventional T cells, as determined by their ability to suppress proliferation of conventional T cells. [Figure 8C] 1 provides a graph showing the suppressive activity of regulatory T cells expressing a TCR of the present disclosure on conventional T cells, as determined by their ability to suppress proliferation of conventional T cells. [Figure 9] 1 provides a graph showing the suppressive activity of regulatory T cells expressing the TCRs of the present disclosure on CD8 T cells and dendritic cells. [Figure 10] 1 provides a graph showing the results of X-scan analysis on TCRs of the present disclosure demonstrating target specificity. [Figure 11] 1 provides a graph showing the suppressive activity of regulatory T cells expressing the TCRs of the present disclosure on conventional T cells. [Figure 12] Graphs are provided showing that Jurkat cells transduced with a TCR of the present disclosure exhibited upregulation of CD69 only when co-cultured with cells expressing HLA-DRB*04:01 (e.g., did not exhibit upregulation when co-cultured with cells that did not express HLA-DRB*04:01). [Figure 13] 1 shows the percent TSDR hypomethylation of six different runs produced by the method described in Example 6. DETAILED DESCRIPTION OF THE INVENTION

[0093] In some aspects, provided herein is an autologous T cell (e.g., regulatory T cell) therapy for the treatment of type 1 diabetes (T1D), e.g., for the treatment of T1D in patients with the HLA-DRB1*04:01 genetic haplotype. In some embodiments, the therapy is a targeted therapy for T1D patients with residual / residual beta cell function in the pancreas (e.g., still producing insulin). In some embodiments, the patient's own regulatory T cells are minimally engineered to express a TCR that specifically recognizes an immunogenic protein fragment of an autoantigen that drives the development of effector T cells in the pancreas and draining lymph nodes, thereby resulting in beta cell destruction. Engineering these regulatory T cells has been shown to enhance the therapeutic potential of the cells, as evidenced by increased retention, activation, proliferation, and differentiation of regulatory T cells when they encounter their cognate antigen-major histocompatibility complex (MHC) (e.g., MHC class II antigen) in the pancreas, e.g., in inflamed islets of Langerhans and pancreatic-draining lymph nodes. In some embodiments, the therapies described herein provide a strong safety profile and highly localized anti-inflammatory effects at the site of disease. In some aspects, the engineered regulatory T cells provided herein (which express a TCR of the present disclosure) exhibit antigen specificity, dose-dependent regulatory T cell functionality, anti-inflammatory cytokine production, and suppression of the production of inflammatory factors (e.g., T cell-derived cytokines). In some embodiments, the engineered regulatory T cells also retain their regulatory T cell phenotype under inflammatory conditions (e.g., in the presence of one or more inflammatory cytokines).

[0094] The present disclosure provides compositions of T cell receptors (TCRs), compositions of engineered regulatory T cells comprising the TCRs, and methods of using the engineered regulatory T cells for the treatment of type 1 diabetes (e.g., stage 3 type 1 diabetes). While 40% of Caucasians in the United States have the HLA-DR3 or -DR4 allele, at least one of these alleles is present in 95% of patients with type 1 diabetes. Diabetes risk is also influenced by the DRB1*04 variant, with DRB1*04:01 having an odds ratio of 8.4. The TCRs of the present disclosure are capable of specifically binding to a proinsulin peptide presented by an MHC molecule, e.g., an MHC class II antigen, e.g., HLA-DRB1, e.g., HLA-DRB1*04:01, with nanomolar to low micromolar binding affinity. Engineering regulatory T cells to express the TCRs of the present disclosure allows the T cells to specifically target distinct cell types and tissues associated with type 1 diabetes to prevent immune-mediated destruction, restore homeostasis, and promote repair in affected tissues.

[0095] While many of the aspects and embodiments described herein are directed to regulatory T cells, the TCRs of the present disclosure may be expressed in other T cell types, depending on the intended use of the T cell expressing the TCR (e.g., use of the T cell to target a preproinsulin peptide, e.g., preproinsulin peptide 73-90).

[0096] T cell receptor In some embodiments, the present disclosure provides T cell receptors (TCRs) that enable antigen-specific targeting (e.g., by engineered regulatory T cells). TCRs are (include) transmembrane heterodimers that comprise an alpha chain and a beta chain linked by a disulfide bond. Within these chains are complementary determining regions (CDRs) that determine the target peptide to which the TCR binds. TCRs activate T cells in which they reside, resulting in an exaggerated immune response. Antigen-presenting cells digest specific proteins (antigens) and present their fragments (peptides) on major histocompatibility complexes (MHC). This peptide-MHC (pMHC) complex binds to the TCR, and other costimulatory molecules are activated, resulting in T cell activation, proliferation, differentiation, apoptosis, and / or cytokine release.

[0097] A TCR specifically binds to a target peptide (e.g., a proinsulin peptide) complexed with an MHC molecule. A TCR is considered to "specifically" bind to a target peptide complexed with an MHC molecule if it has a higher binding affinity to the target peptide complexed with an MHC molecule relative to a non-target peptide complexed with an MHC molecule. A TCR binds to a target peptide complexed with an MHC molecule with at least 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M or 10 -10 M (e.g., 10 -4 M~10 -10M). In some embodiments, a TCR is considered to "specifically" bind a target peptide complexed with an MHC molecule if T cells expressing the TCR are activated (e.g., as assessed by increased CD69 expression) upon contact with the target peptide complexed with an MHC molecule, or are more highly activated toward a non-target peptide complexed with an MHC molecule. In some embodiments, the peptide is presented by a cell expressing the MHC molecule. In some embodiments, the TCR specifically binds a proinsulin peptide (e.g., a preproinsulin peptide). In some embodiments, the TCR specifically binds a preproinsulin 73-90 peptide (e.g., comprising the amino acid sequence of SEQ ID NO: 109). Preproinsulin is an islet autoantigen found in subjects with type 1 diabetes and is abundantly and selectively expressed by pancreatic beta cells.

[0098] In some embodiments, the TCR specifically binds to preproinsulin 73-90 complexed with an MHC molecule in the pancreas of a subject. In some embodiments, the TCR specifically binds to preproinsulin 73-90 complexed with an MHC molecule in the draining lymph nodes of a subject. Lymph node drainage is the process by which the lymphatic system moves lymph to lymph nodes throughout the body.

[0099] In some embodiments, the TCR comprises one or more amino acid sequences set forth in Table 1 (e.g., one or more amino acid sequences belonging to any one of TCR-A, TCR-B, TCR-C, TCR-D, TCR-E, TCR-F, TCR-G, TCR-H, TCR-I, TCR-J, TCR-K, TCR-L, TCR-M, TCR-N, TCR-O, TCR-P, TCR-Q, TCR-R, TCR-S, TCR-T, TCR-U, TCR-V, TCR-X, TCR-Y, and TCR-Z).

[0100] The TCR may comprise any alpha chain CDR1, CDR2, or CDR3 amino acid sequence provided in Table 1. In some embodiments, the alpha CDR1 of the TCR is any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, or 250. In some embodiments, the alpha CDR2 of the TCR is any one of SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, or 251. In some embodiments, the alpha CDR3 of the TCR is any one of SEQ ID NOs: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, or 252. The TCR may comprise any beta chain CDR1, CDR2, or CDR3 amino acid sequence provided in Table 1. In some embodiments, the beta CDR1 of the TCR is any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, or 255. In some embodiments, the beta CDR2 of the TCR is any one of SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, or 256. In some embodiments, the beta CDR3 of the TCR is any one of SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, or 257.

[0101] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 113, 123, 133, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233, 243, or 253. In some embodiments, the beta chain variable region of the TCR comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 9, 19, 29, 39, 49, 59, 69, 79, 89, 99, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, or 258. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, or 308. In some embodiments, the beta chain of the TCR comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 119, 129, 139, 149, 159, 169, 179, 189, 199, 209, 219, 229, 239, 249, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, or 309. In some embodiments, the TCR alpha or beta variable region comprises a V region and a J region. In some embodiments, the TCR beta variable region comprises a V region, a D region, and a J region.

[0102] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 4, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 9. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 5, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 10. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 260, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 261.

[0103] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 14, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 19. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 15, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 20. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 262, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 263.

[0104] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 24, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 29. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 25, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 30. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 264, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 265.

[0105] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 34, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 39. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 35, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 40. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 266, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 267.

[0106] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 44, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 49. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 45, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 50. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 268, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 269.

[0107] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 54, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 59. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 55, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 60. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 270, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 271.

[0108] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 64, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 69. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 65, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 70. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 272, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 273.

[0109] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 74, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 79. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 75, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 80. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 274, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 275.

[0110] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 84, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 89. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 85, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 90. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 276, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 277.

[0111] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 94, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 99. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 95, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 100. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 278, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 279.

[0112] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 113, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 118. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 114, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 119. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 280, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 281.

[0113] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 123, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 128. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 124, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 129. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 282, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 283.

[0114] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 133, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 134, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 139. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 284, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 285.

[0115] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 143, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 148. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 144, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 149. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 286, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 287.

[0116] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 153, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 158. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 154, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 159. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 288, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 289.

[0117] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 163, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 168. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 164, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 169. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 290, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 291.

[0118] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 173, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 178. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 174, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 179. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 292, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 293.

[0119] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 183, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 188. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 184, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 189. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 294, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 295.

[0120] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 193, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 198. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 194, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 19. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 296, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 297.

[0121] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 203, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 208. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 204, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 209. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 298, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 299.

[0122] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 213, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 218. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 214, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 219. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 300, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 301.

[0123] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 223, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 228. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 224, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 229. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 302, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 303.

[0124] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 233, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 238. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 234, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 239. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 304, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 305.

[0125] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 243, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 248. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 244, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 249. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 306, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 307.

[0126] In some embodiments, the alpha chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 253, and the beta chain variable region of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 258. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 254, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 259. In some embodiments, the alpha chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 308, and the beta chain of the TCR comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 309.

[0127] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 4 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 9 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 5 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 10 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 260 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 261 (e.g., TCRb full-length protein).

[0128] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3) and an amino acid sequence (e.g., full-length TCRa) that has at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 318 or 319. In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3) and an amino acid sequence (e.g., full-length TCRb) that has at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 320, 321, 322, 323, 324, or 325. In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 318 (e.g., full-length TCRa), the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 321 or 322 (e.g., full-length TCRb). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 5 (e.g., full-length TCRa), the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 320 (e.g., full-length TCRb). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 319 (e.g., full-length TCRa), the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 324 or 325 (e.g., full-length TCRb).In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 260 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90%, at least 95%, or at least 98% identity to the amino acid sequence of SEQ ID NO: 323 (e.g., TCRb full-length protein).

[0129] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 4 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 9 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 5 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 10 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 260 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 261 (e.g., TCRb full-length protein).

[0130] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 4 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 9 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 5 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 10 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 1-3 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 260 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 6-8 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 261 (e.g., TCRb full-length protein).

[0131] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 14 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 19 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 15 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 20 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 262 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 263 (e.g., TCRb full-length protein).

[0132] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 14 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 19 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 15 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 20 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 262 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 263 (e.g., TCRb full-length protein).

[0133] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 14 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 19 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 15 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 20 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 11-13 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 262 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 16-18 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 263 (e.g., TCRb full-length protein).

[0134] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 24 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 29 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 25 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 30 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 264 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 265 (e.g., TCRb full-length protein).

[0135] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 24 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 29 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 25 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 30 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 264 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 265 (e.g., TCRb full-length protein).

[0136] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 24 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 29 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 25 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 30 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 21-23 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 264 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 26-28 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 265 (e.g., TCRb full-length protein).

[0137] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 34 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 39 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 35 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 40 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 266 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 267 (e.g., TCRb full-length protein).

[0138] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 34 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 39 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 35 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 40 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 266 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 267 (e.g., TCRb full-length protein).

[0139] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 34 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 39 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 35 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 40 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 31-33 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 266 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 36-38 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 267 (e.g., TCRb full-length protein).

[0140] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 44 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 46-48 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 49 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 45 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 50 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 268 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 269 (e.g., TCRb full-length protein).

[0141] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 44 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 46-48 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 49 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 45 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 50 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 268 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 269 (e.g., TCRb full-length protein).

[0142] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 44 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 46-48 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 49 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 45 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 50 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 268 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 41-43 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 269 (e.g., TCRb full-length protein).

[0143] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 54 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 55 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 59 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 60 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 270 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 271 (e.g., TCRb full-length protein).

[0144] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 54 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 55 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 59 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 60 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 270 (e.g., the TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 271 (e.g., the TCRb full-length protein).

[0145] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 54 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 55 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 59 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 60 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 270 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 51-53 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 271 (e.g., TCRb full-length protein).

[0146] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 64 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 66-68 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 69 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 65 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 70 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 272 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 273 (e.g., TCRb full-length protein).

[0147] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 64 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 66-68 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 69 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 65 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 70 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 272 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 273 (e.g., TCRb full-length protein).

[0148] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 64 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 66-68 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 69 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 65 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 70 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 272 (e.g., the TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 61-63 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 273 (e.g., the TCRb full-length protein).

[0149] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 74 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 79 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 75 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 80 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 274 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 275 (e.g., TCRb full-length protein).

[0150] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 74 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 79 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 75 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 80 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 274 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 275 (e.g., TCRb full-length protein).

[0151] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 74 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 79 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 75 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 80 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 71-73 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 274 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 76-78 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 275 (e.g., TCRb full-length protein).

[0152] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 84 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 89 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 85 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 90 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 276 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 277 (e.g., TCRb full-length protein).

[0153] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 84 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 89 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 85 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 90 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 276 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 277 (e.g., TCRb full-length protein).

[0154] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 84 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 89 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 85 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 90 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 81-83 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 276 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 86-88 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 277 (e.g., TCRb full-length protein).

[0155] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 94 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 99 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 95 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 100 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 278 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 279 (e.g., TCRb full-length protein).

[0156] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 94 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 99 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 95 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 100 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 278 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 279 (e.g., TCRb full-length protein).

[0157] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 94 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 99 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 95 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 100 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 91-93 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 278 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 96-98 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 279 (e.g., TCRb full-length protein).

[0158] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 113 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 118 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 114 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 119 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 280 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 281 (e.g., TCRb full-length protein).

[0159] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 113 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 118 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 114 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 119 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 280 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 281 (e.g., TCRb full-length protein).

[0160] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 113 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 118 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 114 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 119 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 110-112 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 280 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 115-117 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 281 (e.g., TCRb full-length protein).

[0161] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 123 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 128 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 124 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 129 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 282 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 283 (e.g., TCRb full-length protein).

[0162] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 123 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 128 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 124 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 129 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 282 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 283 (e.g., TCRb full-length protein).

[0163] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 123 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 128 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 124 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 129 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 120-122 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 282 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 125-127 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 283 (e.g., TCRb full-length protein).

[0164] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 133 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 138 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 134 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 139 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 284 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 285 (e.g., TCRb full-length protein).

[0165] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 133 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 138 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 134 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 139 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 284 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 285 (e.g., TCRb full-length protein).

[0166] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 133 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 138 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 134 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 139 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 130-132 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 284 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 135-137 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 285 (e.g., TCRb full-length protein).

[0167] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 143 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 148 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 144 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 149 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 286 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 287 (e.g., TCRb full-length protein).

[0168] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 143 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 148 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 144 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 149 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 286 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 287 (e.g., TCRb full-length protein).

[0169] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 143 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 148 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 144 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 149 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 140-142 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 286 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 145-147 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 287 (e.g., TCRb full-length protein).

[0170] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 153 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 158 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 154 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 159 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 288 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 289 (e.g., TCRb full-length protein).

[0171] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 153 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 158 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 154 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 159 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 288 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 289 (e.g., TCRb full-length protein).

[0172] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 153 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 158 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 154 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 159 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 150-152 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 290 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 155-157 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 291 (e.g., TCRb full-length protein).

[0173] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 163 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 168 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 164 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 169 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 290 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 291 (e.g., TCRb full-length protein).

[0174] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 163 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 168 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 164 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 169 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 290 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 291 (e.g., TCRb full-length protein).

[0175] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 163 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 168 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 164 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 169 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 160-162 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 290 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 165-167 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 291 (e.g., TCRb full-length protein).

[0176] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 173 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 178 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 174 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 179 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 292 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 293 (e.g., TCRb full-length protein).

[0177] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 173 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 178 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 174 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 179 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 292 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 293 (e.g., TCRb full-length protein).

[0178] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 173 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 178 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 174 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 179 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 170-172 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 292 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 175-177 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 293 (e.g., TCRb full-length protein).

[0179] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 183 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 188 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 294 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 295 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 184 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 189 (e.g., TCRb full-length protein).

[0180] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 183 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 188 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 294 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 295 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 184 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 189 (e.g., TCRb full-length protein).

[0181] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 183 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 188 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 294 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 295 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 180-182 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 184 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 185-187 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 189 (e.g., TCRb full-length protein).

[0182] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 193 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 198 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 296 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 297 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 194 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 199 (e.g., TCRb full-length protein).

[0183] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 193 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 198 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 296 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 297 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 194 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 199 (e.g., TCRb full-length protein).

[0184] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 193 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 198 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 296 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 297 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 190-192 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 194 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 195-197 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 199 (e.g., TCRb full-length protein).

[0185] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 203 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 208 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 298 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 299 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 204 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 209 (e.g., TCRb full-length protein).

[0186] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 203 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 208 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 298 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 299 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 204 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 209 (e.g., TCRb full-length protein).

[0187] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 203 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 208 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 298 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 299 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 200-202 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 204 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 205-207 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 209 (e.g., TCRb full-length protein).

[0188] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 213 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 218 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 300 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 301 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 214 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 219 (e.g., TCRb full-length protein).

[0189] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 213 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 218 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 300 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 301 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 214 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 219 (e.g., TCRb full-length protein).

[0190] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 213 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 218 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 300 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 301 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 210-212 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 214 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 215-217 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 219 (e.g., TCRb full-length protein).

[0191] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 223 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 228 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 302 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 303 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 224 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 229 (e.g., TCRb full-length protein).

[0192] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 223 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 228 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 302 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 303 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 224 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 229 (e.g., TCRb full-length protein).

[0193] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 223 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 228 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 302 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 303 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 220-222 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 224 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 225-227 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 229 (e.g., TCRb full-length protein).

[0194] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 233 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 238 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 304 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 305 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 234 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 239 (e.g., TCRb full-length protein).

[0195] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 233 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 238 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 304 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 305 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 234 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 239 (e.g., TCRb full-length protein).

[0196] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 233 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 238 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 304 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 305 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 230-232 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 234 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 235-237 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 239 (e.g., TCRb full-length protein).

[0197] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 243 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 248 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 306 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 307 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 244 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 249 (e.g., TCRb full-length protein).

[0198] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 243 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 248 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 306 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 307 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 244 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 249 (e.g., TCRb full-length protein).

[0199] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 243 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 248 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 306 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 307 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 240-242 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 244 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 245-247 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 249 (e.g., TCRb full-length protein).

[0200] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 253 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 258 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDR1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 308 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 309 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 90% identity to SEQ ID NO: 254 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 90% identity to SEQ ID NO: 259 (e.g., TCRb full-length protein).

[0201] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 253 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 258 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDR1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 308 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 309 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 254 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 259 (e.g., TCRb full-length protein).

[0202] In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 253 (e.g., TCRa variable domain), an amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 258 (e.g., TCRb variable domain). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDR1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 308 (e.g., TCRa full-length protein), an amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDR1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 309 (e.g., TCRb full-length protein). In some embodiments, the TCR comprises the amino acid sequence of SEQ ID NOs: 250-252 (e.g., TCRa CDRs 1-3), an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 254 (e.g., TCRa full-length protein), the amino acid sequence of SEQ ID NOs: 255-257 (e.g., TCRb CDRs 1-3), and an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 259 (e.g., TCRb full-length protein).

[0203] Regulatory T cells (or other T cells) can comprise any one or more of the TCRs or TCR components provided in Table 1 (e.g., TCRa CDR1, TCRa CDR2, TCRa CDR3, TCRa variable domain, TCRa complete, TCRa complete (DS), TCRb CDR1, TCRb CDR2, TCRb CDR3, TCRb variable domain, TCRb complete, TCRb complete (DS), and / or complete ORF). In some embodiments, the TCR further comprises a ribosomal skip P2A element (e.g., SEQ ID NO: 103). In some embodiments, the orientation of the TCR is beta chain-P2A-alpha chain. In some embodiments, the orientation of the TCR is alpha chain-P2A-beta chain. Table 1-1 Table 1-2 Table 1-3 Table 1-4 Table 1-5 Table 1-6 Table 1-7 Table 1-8 Table 1-9 Table 1-10 Table 1-11 Table 1-12 Table 1-13 Table 1-14 Table 1-15 Table 1-16 Table 1-17 [Table 1-18] [Table 1-19] [Table 1-20] [Table 1-21] [Table 1-22] [Table 1-23] [Table 1-24]

[0204] In some embodiments, the TCR is an exogenous TCR. An exogenous TCR can be any TCR introduced into a T cell, e.g., a regulatory T cell, where the TCR is not exogenous to the regulatory T cell (i.e., naturally occurring in the regulatory T cell). For example, in some embodiments, the exogenous TCR is encoded by a nucleic acid that is not exogenous to the regulatory T cell (i.e., not naturally occurring in the genome of the regulatory T cell). In some embodiments, the nucleic acid is an engineered nucleic acid, e.g., a recombinant or synthetic nucleic acid. Thus, an exogenous TCR may also be referred to herein as an engineered TCR. Regulatory T cells comprising (e.g., expressing) an engineered TCR are considered to be engineered regulatory T cells.

[0205] The TCR can be a human TCR (comprises human TCR nucleic acid and / or amino acid sequences). In some embodiments, the TCR is a TCR from a monkey, mouse, rat, or any other animal. In some embodiments, the TCR is chimeric (comprises nucleic acid and / or amino acid sequences from two or more different species).

[0206] The target peptide can be a peptide or portion of a peptide associated with type 1 diabetes. The target peptide associated with type 1 diabetes can be a peptide belonging to proinsulin. In some embodiments, the target peptide is a peptide that is overexpressed in a population of cells associated with type 1 diabetes relative to a control (e.g., relative to a population of cells not associated with type 1 diabetes). In some embodiments, the target peptide is a peptide that is expressed (or expressed at a higher level) at a site (e.g., tissue) of disease relative to other sites in the body.

[0207] In some embodiments, the target protein is a proinsulin peptide. As described herein, the proinsulin peptide is a peptide of proinsulin, preproinsulin (PPI), or proinsulin C. In some embodiments, the proinsulin peptide comprises the sequence GAGSLQPLALEGSLQKRG (SEQ ID NO: 109). In some embodiments, the proinsulin peptide consists of the sequence of SEQ ID NO: 109. Thus, in some embodiments, the TCR of the present disclosure specifically binds to a peptide comprising or consisting of the sequence of SEQ ID NO: 109.

[0208] Preproinsulin (PPI) is a 110-amino acid protein used to synthesize insulin. PPI is an inactive precursor molecule that is directly translated into the rough endoplasmic reticulum (RER), where its signal peptide is removed by signal peptidases to form proinsulin. Proinsulin is the penultimate precursor of insulin and is produced in pancreatic beta cells. In humans, the proinsulin peptide is approximately 86 amino acid residues long and is formed by three distinct chains (A chain, B chain, and C peptide). Proinsulin is post-translationally cleaved into three peptides: the B and A chain peptides, which are covalently linked via two disulfide bonds to form insulin, and the C peptide. Proinsulin is cleaved by proprotein convertase 1 / 3 and proprotein convertase 2, which remove the C peptide, and carboxypeptidase E, which removes two paired amino acids from the terminus of the protein, resulting in active insulin. Interestingly, proinsulin demonstrates affinity for the insulin receptor. Proinsulin C-peptide ("C-peptide") is approximately 31 amino acids long. C-peptide is cleaved from proinsulin during its conversion to insulin. The full-length C-peptide is amino acids 33-63 of proinsulin.

[0209] In some embodiments, the target peptide is subject to an increased autoimmune response in subjects with type 1 diabetes relative to controls. In some embodiments, the target peptide is a peptide present at a site associated with type 1 diabetes in a subject relative to a non-affected site in the same subject. In some embodiments, the target peptide is specifically presented by an MHC allele associated with the presence of type 1 diabetes in a subject.

[0210] In some embodiments, the target peptide is a peptide that is overexpressed in cells of a subject with type 1 diabetes relative to a control (e.g., relative to a healthy subject). A target peptide is considered to be overexpressed in a cell (e.g., one associated with type 1 diabetes) if expression of the target peptide in the cell is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% higher than control cells (e.g., a population of cells not associated with type 1 diabetes) in the cell. In some embodiments, a target peptide is overexpressed in cells of a subject with type 1 diabetes if expression of the target peptide in cells of the subject with type 1 diabetes is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% higher in the subject with type 1 diabetes relative to a healthy subject (e.g., a subject without type 1 diabetes).

[0211] In some embodiments, the target peptide is a peptide that is highly expressed in cells at a site of disease in a subject with type 1 diabetes relative to a control (e.g., target peptide expression in a non-diseased / non-diseased site in the subject). A target peptide is considered to be highly expressed in cells at a site of disease if expression of the target peptide in disease site cells (e.g., those associated with type 1 diabetes) is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% higher than control cells (e.g., cells not present at the site of disease).

[0212] In some embodiments, the target peptide is a peptide complexed with MHC having an HLA haplotype associated with type 1 diabetes.

[0213] In some embodiments, the TCR is (or is encoded as) a single polypeptide (e.g., comprising a beta chain and an alpha chain). In some embodiments, the TCR comprises an N-terminal beta chain and a C-terminal alpha chain. In other embodiments, the TCR comprises an N-terminal alpha chain and a C-terminal beta chain.

[0214] The TCR may comprise a linker domain located between the alpha chain and the beta chain. In some embodiments, the linker domain comprises a self-cleaving peptide sequence (e.g., a self-cleaving peptide sequence located between the alpha chain and the beta chain). A self-cleaving peptide sequence is a peptide sequence that induces separation of a polypeptide into two peptides using a non-classical mechanism. In some embodiments, the self-cleaving peptide sequence may induce ribosomal skipping during translation of the polypeptide. In some embodiments, the self-cleaving peptide sequence is 10-30, 10-25, 15-30, 15-25, or 18-22 amino acids in length. In some embodiments, the self-cleaving peptide sequence may be a 2A peptide sequence. The 2A peptide sequence may comprise, for example, the DXEXNPGP (SEQ ID NO: 101) amino acid motif, in which X can be any amino acid. In some embodiments, the 2A peptide sequence is a P2A (derived from porcine teschovirus-1 2A), E2A (derived from equine rhinitis A virus), F2A (derived from foot-and-mouth disease virus), or T2A (derived from Thosea asigna virus 2A) peptide sequence. The T2A peptide sequence may, for example, comprise the amino acid sequence of EGRGSLLTCGDVEENPGP (SEQ ID NO: 102). The P2A peptide sequence may, for example, comprise the amino acid sequence of ATNFSLLKQAGDVEENPGP (SEQ ID NO: 103). The E2A peptide sequence may, for example, comprise the amino acid sequence of QCTNYALLKLAGDVESNPGP (SEQ ID NO: 104). The F2A peptide sequence may, for example, comprise the amino acid sequence of VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 105).

[0215] In some embodiments, a TCR comprises two or more polypeptides, for example, in some embodiments, a TCR comprises a first polypeptide comprising an alpha chain and a second polypeptide comprising a beta chain.

[0216] In some embodiments, the TCR comprises one or more cysteine ​​residues present in the alpha chain of the TCR, which are capable of forming one or more disulfide bonds with one or more cysteine ​​residues in the beta chain of the TCR. In some embodiments, the exogenous TCR comprises one or more cysteine ​​residues present in the alpha chain constant region of the TCR, which form or are capable of forming one or more disulfide bonds with one or more cysteine ​​residues in the beta chain constant region of the TCR.

[0217] In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 48 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., T48C). In some embodiments, the TCR beta chain constant region comprises an amino acid substitution at position 57 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., S57C) amino acid substitution. In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 48 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., T48C), and the TCR beta chain constant region comprises an amino acid substitution at position 57 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., S57C) amino acid substitution, wherein the cysteine ​​residue at position 48 of the alpha chain is capable of forming a disulfide bond with the cysteine ​​residue at position 57 of the beta chain.

[0218] In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 45 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., T45C). In some embodiments, the TCR beta chain constant region comprises an amino acid substitution at position 77 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., S77C) amino acid substitution. In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 45 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., T45C), and the TCR beta chain constant region comprises an amino acid substitution at position 77 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., S77C) amino acid substitution, wherein the cysteine ​​residue at position 45 of the alpha chain is capable of forming a disulfide bond with the cysteine ​​residue at position 77 of the beta chain.

[0219] In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 10 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., Y10C). In some embodiments, the TCR beta chain constant region comprises an amino acid substitution at position 17 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., S17C) amino acid substitution. In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 10 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., Y10C), and the TCR beta chain constant region comprises an amino acid substitution at position 17 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., S17C) amino acid substitution, wherein the cysteine ​​residue at position 10 of the alpha chain is capable of forming a disulfide bond with the cysteine ​​residue at position 17 of the beta chain.

[0220] In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 45 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., T45C). In some embodiments, the TCR beta chain constant region comprises an amino acid substitution at position 59 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., D59C) amino acid substitution. In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 45 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., T45C), and the TCR beta chain constant region comprises an amino acid substitution at position 59 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., D59C) amino acid substitution, wherein the cysteine ​​residue at position 45 of the alpha chain is capable of forming a disulfide bond with the cysteine ​​residue at position 59 of the beta chain.

[0221] In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 15 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., S15C). In some embodiments, the TCR beta chain constant region comprises an amino acid substitution at position 15 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., E15C) amino acid substitution. In some embodiments, the TCR alpha chain constant region comprises an amino acid substitution at position 15 relative to a TCR alpha chain constant region comprising the amino acid sequence of SEQ ID NO: 106, introducing a cysteine ​​(e.g., S15C), and the TCR beta chain constant region comprises an amino acid substitution at position 15 relative to a TCR beta chain constant region comprising the amino acid sequence of SEQ ID NO: 108, introducing a cysteine ​​(e.g., E15C) amino acid substitution, wherein the cysteine ​​residue at position 15 of the alpha chain is capable of forming a disulfide bond with the cysteine ​​residue at position 15 of the beta chain.

[0222] Nucleic acid encoding a TCR In some embodiments, the disclosure provides a nucleic acid encoding a TCR (e.g., a TCR listed in Table 1). The nucleic acid can be or include deoxyribonucleic acid (DNA), ribonucleic acid (RNA) (e.g., messenger RNA), threose nucleic acid (TNA), glycol nucleic acid (GNA), peptide nucleic acid (PNA), locked nucleic acid (LNA), ethylene nucleic acid (ENA), cyclohexenyl nucleic acid (CeNA), and / or chimeras.

[0223] As used herein, a nucleic acid generally refers to an engineered nucleic acid. An engineered nucleic acid is a non-naturally occurring polynucleotide (e.g., at least two nucleotides covalently linked to each other, optionally containing phosphodiester bonds, referred to as a phosphodiester backbone). Engineered nucleic acids include recombinant nucleic acids and synthetic nucleic acids. A recombinant nucleic acid is a molecule constructed by joining nucleic acids (e.g., isolated nucleic acids, synthetic nucleic acids, or a combination thereof) from two different organisms (e.g., human and mouse). A synthetic nucleic acid is a molecule that is amplified or synthesized by chemical or other means. Synthetic nucleic acids include those that are chemically or otherwise modified but are still capable of base pairing (binding) with naturally occurring nucleic acid molecules. Recombinant and synthetic nucleic acids also include molecules resulting from replication of any of the above.

[0224] The engineered nucleic acids of the present disclosure can be produced using standard molecular biology methods (see, e.g., Green and Sambrook, Molecular Cloning, A Laboratory Manual, 2012, Cold Spring Harbor Press). In some embodiments, the nucleic acids are produced using GIBSON ASSEMBLY® cloning (see, e.g., Gibson, D.G.E. al. Nature Methods, 343-345, 2009, and Gibson, D.G.E. al. Nature Methods, 901-903, 2010, each of which is incorporated herein by reference). GIBSON ASSEMBLY® typically uses three enzymatic activities in a single-tube reaction: a 5' exonuclease, a 3' extension activity of a DNA polymerase, and a DNA ligase activity. The 5' exonuclease activity chews back the 5' terminal sequence, exposing complementary sequences for annealing. Polymerase activity then fills in the gaps on the annealed domains. DNA ligase then seals the nicks, covalently linking the DNA fragments to each other. The overlapping sequences of adjacent fragments are much longer than those used in Golden Gate Assembly, thus resulting in a higher percentage of correct assembly. The MegaGate molecular cloning method may also be used. MegaGate is a toxin-free Gateway technology that eliminates the ccdb toxin used in Gateway recombinase cloning and instead uses meganuclease-mediated digestion to eliminate background vectors during cloning (see, for example, Kramme C. et al. STAR Protoc. 2021 Oct. 22;2(4):100907, which is incorporated herein by reference). Other methods of producing engineered polynucleotides may also be used in accordance with the present disclosure.

[0225] In some embodiments, the engineered nucleic acid comprises a promoter operably linked to an open reading frame. A promoter is (includes) a nucleotide sequence (e.g., ATG) to which RNA polymerase binds for initial transcription. A promoter is typically located directly upstream (at its 5' end) of the transcription start site. In some embodiments, the promoter is a heterologous promoter. A heterologous promoter is not naturally associated with the open reading frame to which it is operably linked. In some embodiments, the promoter is an inducible promoter. Inducible promoters can be regulated in vivo, for example, by chemical agents, temperature, or light. Non-limiting examples of promoters that can be used as provided herein include the Eukaryotic Translation Elongation Factor 1 alpha (EF-1 alpha) promoter and the MND promoter (myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer binding site replaced) (see, e.g., Gill, D.R. et al. Gene Ther. 2001;8:1539-46, and Astrakhan, A. et al. Blood 2012;119:4395-4407).

[0226] An open reading frame is (inclusive) a contiguous series of codons that begins with a start codon (e.g., ATG) and ends with a stop codon (e.g., TAA, TAG, or TGA) that encodes a polypeptide, e.g., a protein. An open reading frame is operably linked to a promoter if the promoter controls transcription of the open reading frame.

[0227] The nucleic acid encoding the TCR can be a vector or a plasmid. In some embodiments, the vector is a viral vector. For example, the vector can be a lentiviral vector, an adenoviral vector, an adeno-associated viral (AAV) vector, a herpesvirus vector, a retroviral vector, or a baculoviral vector. Viral vectors provide efficient delivery of exogenous TCRs to the regulatory T cells of the present disclosure. Exemplary viral vectors can be derived from double-stranded DNA viruses, including lentiviruses, retroviruses (e.g., Retroviridae viral vectors), adenoviruses (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvoviruses (e.g., adeno-associated viruses), coronaviruses, negative-strand RNA viruses such as orthomyxoviruses (e.g., influenza viruses), rhabdoviruses (e.g., rabies and vesicular stomatitis viruses), paramyxoviruses (e.g., measles and Sendai), positive-strand RNA viruses such as picornaviruses and alphaviruses, as well as adenoviruses, herpesviruses (e.g., herpes simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus, replication-deficient herpesviruses), and poxviruses (e.g., vaccinia, Modified Vaccinia Ankara (MVA), fowlpox, and canarypox). Other viruses include, for example, Norwalk virus, togavirus, flavivirus, reovirus, papovavirus, hepadnavirus, human papillomavirus, human foamy virus, and hepatitis virus.Examples of retroviruses include avian leukosis-sarcoma, avian C virus, mammalian C, B, and D viruses, oncoretroviruses, HTLV-BLV group, alpharetroviruses, gammaretroviruses, Spumaretrovirus genus, murine leukemia virus, murine sarcoma virus, mouse mammary tumor virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, gibbon leukemia virus, Mason-Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus, and lentiviruses.

[0228] In some embodiments, the nucleic acid encoding the exogenous TCR is RNA (e.g., messenger RNA (mRNA)). In some embodiments, the mRNA comprises a 5' cap, a 5' untranslated region (UTR), an ORF, a 3' UTR, and / or a poly(A) tail.

[0229] In some embodiments, the nucleic acid is codon-optimized. Codon optimization methods are known in the art. In some embodiments, codon optimization can be used to match codon frequencies in the target organism and the host organism to ensure proper folding, bias the GC content to increase RNA (e.g., mRNA) stability or reduce secondary structures, minimize tandemly repeated codons or base runs that may impair gene assembly or expression, customize transcriptional and translational control regions, insert or remove protein transport sequences, remove / add post-translational modification sites (e.g., glycosylation sites) in the encoded protein, add, remove, or shuffle protein domains, insert or delete restriction sites, modify ribosome binding sites and RNA (e.g., mRNA) degradation sites, adjust the translation rate to ensure proper folding of various domains of the protein, or reduce or eliminate problematic secondary structures within the polynucleotide.

[0230] In some embodiments, the nucleic acid encoding a TCR comprises a promoter operably linked to a coding sequence encoding an exogenous human TCR. The promoter may be a viral promoter or a native promoter. In some embodiments, the promoter is a constitutively active promoter or an inducible promoter. In some embodiments, the promoter is the eukaryotic translation elongation factor 1 alpha (EF-1 alpha) promoter and the MND promoter (myeloproliferative sarcoma virus enhancer, deletion of the negative control region, substitution of the dl587rev primer binding site) (see, e.g., Gill, D.R. et al. Gene Ther. 2001;8:1539-46, and Astrakhan, A. et al. Blood 2012;119:4395-4407).

[0231] The vector may also include a stop codon and / or an expression enhancer element. Any suitable vector, promoter, enhancer, and stop codon known in the art may be used. In some embodiments, the enhancer element is an optimized post-transcriptional regulatory element (oPRE), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). The WPRE can be a wild-type WPRE or a WPRE mutant sequence (e.g., WPRE-mut6, which has a mutant woodchuck hepatitis virus X protein open reading frame translation start site). In some embodiments, the WPRE (e.g., WPRE-mut6) is that described in Zanta-Boussif, MA et al., Gene Therapy volume 16, pages 605-619 (2009).

[0232] In some embodiments, the vector encoding a T cell receptor (TCR) is a recombinant lentiviral vector encoding a TCR. The recombinant lentiviral vector can be a third-generation vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped self-inactivating vector (e.g., generated using a split genome four-plasmid system (rev; gag + pol; VSV-G; gene encoding a TCR)). In some embodiments, the recombinant lentiviral vector encodes any of the TCRs described herein. In some embodiments, the recombinant lentiviral vector encodes a TCR comprising one or more amino acid sequences set forth in Table 1 (e.g., a CDR sequence belonging to any one of TCR-A, TCR-B, TCR-C, TCR-D, TCR-E, TCR-F, TCR-G, TCR-H, TCR-I, TCR-J, TCR-K, TCR-L, TCR-M, TCR-N, TCR-O, TCR-P, TCR-Q, TCR-R, TCR-S, TCR-T, TCR-U, TCR-V, TCR-X, TCR-Y, and TCR-Z). In some embodiments, the recombinant lentiviral vector encodes the beta and alpha chains of any one of the TCRs provided in Table 1 (e.g., TCR-A, TCR-B, TCR-C, TCR-D, TCR-E, TCR-F, TCR-G, TCR-H, TCR-I, TCR-J, TCR-K, TCR-L, TCR-M, TCR-N, TCR-O, TCR-P, TCR-Q, TCR-R, TCR-S, TCR-T, TCR-U, TCR-V, TCR-X, TCR-Y, or TCR-Z), wherein the beta and alpha chains are linked by a ribosomal skipping P2A element.

[0233] In some embodiments, the nucleic acid encoding the TCR comprises a nucleotide sequence having greater than 90%, greater than 95%, or greater than 98% identity to SEQ ID NO: 326. In some embodiments, the nucleic acid encoding the TCR comprises the nucleotide sequence of SEQ ID NO: 326.

[0234] Isolated regulatory T cells The present disclosure provides methods for producing regulatory T cells (e.g., stable regulatory T cells) that express the TCRs described herein, and compositions comprising such regulatory T cells. These regulatory T cells can be used, for example, to treat type 1 diabetes. reg Regulatory T cells, also referred to as "regulatory T cells," are (and include) T cells that regulate the immune system. Regulatory T cells are immunosuppressive and generally suppress or downregulate the induction and proliferation of effector T cells. Regulatory T cells are also known as naive CD4 + They are believed to be derived from the same lineage as the cells and express the biomarkers CD4, CD25, and FOXP3. In some embodiments, the regulatory T cells (e.g., stable regulatory T cells) comprise an exogenous human TCR that specifically binds to proinsulin. This binding occurs when proinsulin is complexed with an MHC molecule (e.g., MHC class I or MHC class II).

[0235] Stable or thymic regulatory T cells are regulatory T cells that contain a hypomethylated T cell-specific demethylation region (TSDR) at the endogenous FOXP3 locus and express CD4, CD25, and FOXP3. Regulatory T cells are considered "stable" (also interchangeably referred to as "phenotypically stable") when they are terminally differentiated, i.e., when they have lost their ability to change their cell fate. During thymic regulatory T cell development, the genome organizer Special AT-rich Sequence-Binding Protein 1 (SATB1) binds to CD4. + CD8 +It binds to specific genomic sites from the thymocyte stage, opening up chromatin and activating super-enhancers associated with many regulatory T cell signature genes, such as FOXP3, IL2RA, (CD25), CTLA4, IKZF2 (HELIOS), and IFZF4 (EOS). SATB1 and myeloid / lymphoid or mixed-lineage leukemia 4 (MLL4), enzymes involved in enhancer priming, occupy a newly identified conserved enhancer region, commonly referred to as Conserved Noncoding Sequence 0 (CNS0), in the FOXP3 locus, subsequently activating enhancers in CNS3 and CNS2, followed by promoter activation. This results in stable hypomethylation and expression of FOXP3 and other regulatory T cell-associated genes, thereby resulting in a stable regulatory T cell phenotype. Epigenetic mechanisms, particularly DNA methylation / demethylation, which is heritable through cell division, play an important role in the stable maintenance of regulatory T cell-specific gene expression. Hypomethylation of the TSDR, an evolutionarily conserved CpG-rich regulatory element of the FOXP3 gene, is associated with FOXP3 expression. The TSDR of the endogenous FOXP3 locus becomes hypomethylated when the methyl group from one or more methylated cytosines in the TSDR is removed and replaced with a cytosine.In some embodiments, measurement of the methylation status of the TSDR of the FOXP3 locus is as described in Kressler et al., "Targeted De-Methylation of the FOXP3-TSDR Is Sufficient to Induce Physiological FOXP3 Expression but Not a Functional Regulatory T Phenotype," Frontiers in Immunology, January 7, 2021, or Schreiber, et al., "The Regulatory T-Specific Demethylated Region Stabilizes Foxp3 Expression Independently of NF-κB Signaling," PLOS One, February 5, 2014. For example, genomic DNA can be isolated and bisulfite conversion performed. In some embodiments, this converts all unmethylated cysteines to uracils. The TSDR region can be amplified independently of methylation status using primers that amplify bisulfite-converted DNA. Two competing probes detect the abundance of methylated or unmethylated CpGs, and the total and / or relative abundance is determined by ddPCR or pyrosequencing. Alternatively, next-generation sequencing (NGS) can be performed on bisulfite-converted DNA to determine the sequence of the bisulfite-converted DNA at the TSDR locus.

[0236] In some embodiments, stable regulatory T cells retain markers of stability in the presence of pro-inflammatory conditions (e.g., in the presence of one or more pro-inflammatory cytokines). In some embodiments, stable regulatory T cells comprise a hypomethylated T-regulatory cell-specific demethylated region (TSDR) at the endogenous FOXP3 locus and express CD4, CD25, and FOXP3 in the presence of pro-inflammatory conditions (e.g., in the presence of one or more pro-inflammatory cytokines). In some embodiments, stable regulatory T cells comprise a hypomethylated T-regulatory cell-specific demethylated region (TSDR) at the endogenous FOXP3 locus and express CD4, CD25, and FOXP3 in the presence of pro-inflammatory conditions for at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 21, 22, 23, 24, or 25 days.

[0237] Stable regulatory T cells expressing an exogenous TCR (e.g., a TCR described in Table 1) that specifically binds to a target peptide (e.g., proinsulin) complexed with an MHC molecule further exhibit one or more of the following functions: (i) cytokine secretion activity (e.g., secretion of IL-10, IL-4, IL-6, IL-11, and / or IL-13) when the TCR contacts the target peptide; (ii) expression of activation markers associated with regulatory T cells (e.g., expression of CD69, 4-1BB, CD25, CD71, and / or CTLA-4) when the TCR contacts the target peptide; and / or (iii) suppressive activity (e.g., the ability of the stable regulatory T cells to suppress the activation of conventional T cells with specificity for a common target peptide) when the TCR contacts the target peptide complexed with an MHC molecule.

[0238] Thus, the regulatory T cells of the isolated cell populations provided herein may comprise a hypomethylated TSDR at the endogenous FOXP3 locus, may comprise expression of CD4, CD25, and FOXP3, and / or may exhibit cytokine secretion, activation, and / or suppressive activity.

[0239] In some embodiments, regulatory T cells (e.g., stable regulatory T cells) are CD25 + In some embodiments, the regulatory T cells (e.g., stable regulatory T cells) are CD25 high As used herein, "CD25 + / high " means that the cells + or CD25 high This means that either CD25 + The cells are those that express detectable levels of CD25 (e.g., detectable by FACS). + / high The cells are those that express CD25 above a threshold or control level. For example, CD25 + In the context of an isolated cell population containing CD25 cells, a subpopulation of these cells may be identified as CD25 + The former subpopulation may express higher levels of CD25 than the other subpopulation of cells (or than the CD25+ cells of all other cells in the population). high The latter subpopulation is thought to be CD25 + Only (CD25 high It is considered that

[0240] In some embodiments, regulatory T cells (e.g., stable regulatory T cells) are CD4 + is.

[0241] In some embodiments, regulatory T cells (e.g., stable regulatory T cells) express CD127 - / lo As used herein, "CD127 - / lo " means that cells express CD127 - or CD127 lo This means that either CD127 - The cells are those that do not express CD127, do not express detectable levels of CD127 (e.g., by FACS), or express CD127 below a threshold or control level. For example, at least some proportion of CD127 +In the context of an isolated cell population containing CD127 cells (e.g., detectable by FACS), a subpopulation of these cells may be identified by the expression of CD127 in this population. + The former subpopulation may express lower levels of CD127 relative to another subpopulation of cells (or relative to all other CD127+ cells in the population). lo (CD127 - It is considered that

[0242] In some embodiments, regulatory T cells (e.g., stable regulatory T cells) are CD45RA + is.

[0243] In some embodiments, the regulatory T cells (e.g., stable regulatory T cells) are FOXP3 + is.

[0244] In some embodiments, regulatory T cells (e.g., stable regulatory T cells) are CD25 + / high CD4 + CD127 - / lo That is, regulatory T cells express CD25 or express it at high levels, express CD4, and do not express CD127 or express it at low levels. In some embodiments, regulatory T cells do not express CD127. In some embodiments, regulatory T cells (e.g., stable regulatory T cells) express CD25 + / high CD4 + CD127 - / lo / CD45RA + Regulatory T cells are cells. Thus, regulatory T cells express CD25 or express high levels of CD25, express CD4 and CD45RA, and do not express or express low levels of CD127. In some embodiments, regulatory T cells express FOXP3. In some embodiments, regulatory T cells express CD25 + / high CD4 + CD127 - / lo FOXP3 +Thus, regulatory T cells express CD25 or express high levels of CD25, CD4 and FOXP3, and do not express CD127 or express low levels of CD127. In some embodiments, regulatory T cells express CD25 + / high CD4 + CD45RA + CD127 - / lo FOXP3 + Thus, regulatory T cells express CD25 or express high levels of CD25, CD4, CD45RA, and FOXP3, and do not express CD127 or express low levels of CD127.

[0245] A cell "expresses" a biomarker (e.g., CD4, CD25, CD45RA, and / or FOXP3) if the biomarker can be detected using a conventional protein expression assay, e.g., an antibody detection assay. An antibody detection assay in this context involves detection of an antibody that binds to the biomarker of interest (e.g., CD4, CD25, CD45RA, or FOXP3) when the biomarker is present inside the cell or on the surface of the cell (e.g., using Fluorescence Activated Cell Sorting (FACS) or Western blot). A cell "does not express" a biomarker (e.g., CD127) if its expression is not detectable using a conventional protein assay. A cell "expresses" a "low level" of a biomarker (e.g., CD127) if its expression is lower than control levels (e.g., at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% lower). In some embodiments, the control level of a biomarker in regulatory T cells is a level of a conventional T cell or CD8 +The expression level of a biomarker (e.g., CD127) in T cells. For example, a regulatory T cell expresses a low level of CD127 if its expression is lower (e.g., at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% lower) than that of CD127 in conventional T cells.

[0246] In some embodiments, the stable regulatory T cells of the isolated cell populations provided herein maintain a hypomethylated TSDR at the endogenous FOXP3 locus over time (i.e., the TSDR at the endogenous FOXP3 locus of viable cells (e.g., in culture, in cryoprotectant, and / or in vivo) is hypomethylated for a certain measurable period of time). For example, the stable regulatory T cells of the isolated cell populations may maintain a hypomethylated TSDR at the endogenous FOXP3 locus for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days (e.g., in culture, in cryoprotectant, and / or in vivo after being obtained from a subject or after being transduced with a nucleic acid expressing a TCR). In some embodiments, stable regulatory T cells maintain hypomethylated TSDR at the endogenous FOXP3 locus for more than 5 days, more than 10 days, more than 15 days, or more than 20 days (e.g., in culture, in cryoprotectant, and / or in vivo after being obtained from a subject or after transduction with a nucleic acid expressing a TCR). In some embodiments, stable regulatory T cells maintain hypomethylated TSDR at the endogenous FOXP3 locus for 1-20 days, 1-10 days, 1-5 days, 5-30 days, 5-20 days, 10-40 days, or 25-50 days (e.g., in culture, in cryoprotectant, and / or in vivo after being obtained from a subject or after transduction with a nucleic acid expressing a TCR).

[0247] In some embodiments, the regulatory T cells of the isolated cell population of the present disclosure are autologous cells. The term autologous in this context refers to cells obtained from the same subject to which the cells are subsequently administered (e.g., after manipulation to introduce exogenous TCR into the cells). For example, a population of cells can be obtained from a subject, subjected to the methods described herein, and then administered to the same subject (the same subject from which the population of cells was originally obtained) to treat type 1 diabetes. In such embodiments, the population of cells administered to the subject comprises autologous regulatory T cells.

[0248] In some embodiments, the regulatory T cell of the present disclosure is allogeneic cell.In this context, the term allogeneic refers to the cell that is obtained from one subject and then administered to another subject.For example, a group of cells can be obtained from a subject, and then be subjected to the method described herein, and then be administered to another subject to treat autoimmune disease.

[0249] In some embodiments, regulatory T cells (eg, a biological sample containing regulatory T cells) are obtained from a subject diagnosed with or suspected of having type 1 diabetes.

[0250] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with a major histocompatibility complex (MHC) molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 2, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 3, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 6, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 7, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 8.

[0251] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 3. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 4. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 260.

[0252] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 6, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 7, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 9. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 10. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 261.

[0253] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 11, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 12, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 13, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 17, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 18.

[0254] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 11, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 12, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 15. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 262.

[0255] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 17, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 18. In some embodiments, the beta chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 19. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 20. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 263.

[0256] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 21, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 22, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 23, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 26, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 27, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 28.

[0257] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 21, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 22, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 23. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 24. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 25. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 264.

[0258] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 26, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 27, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 28. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 29. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 30. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 265.

[0259] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 31, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 32, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 33, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 36, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 37, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 38.

[0260] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 31, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 32, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 33. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 34. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 35. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 266.

[0261] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 36, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 37, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 38. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 39. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 40. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 267.

[0262] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 41, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 42, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 43, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 46, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 47, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 48.

[0263] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 41, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 42, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 43. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 44. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 45. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 268.

[0264] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 46, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 47, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 48. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 49. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 50. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 269.

[0265] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 51, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 52, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 53, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 56, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 57, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 58.

[0266] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 51, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 52, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 53. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 54. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 55. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 270.

[0267] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 56, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 57, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 58. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 59. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 60. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 271.

[0268] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 61, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 62, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 63, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 66, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 67, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 68.

[0269] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 61, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 62, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 63. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 64. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 65. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 272.

[0270] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 66, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 67, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 68. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 69. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 70. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 273.

[0271] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 71, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 72, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 73, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 76, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 77, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 78.

[0272] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 71, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 72, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 73. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 74. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 75. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 274.

[0273] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 76, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 77, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 78. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 79. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 80. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 275.

[0274] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 81, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 82, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 83, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 86, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 87, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 88.

[0275] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 81, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 82, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 83. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 84. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 85. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 276.

[0276] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 86, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 87, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 88. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 89. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 90. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 277.

[0277] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 91, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 92, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 93, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 96, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 97, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 98.

[0278] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 91, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 92, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 93. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 94. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 95. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 278.

[0279] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 96, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 97, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 98. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 99. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 100. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 279.

[0280] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 110, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 111, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 112, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 115, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 116, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 117.

[0281] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 110, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 111, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 112. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 113. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 280.

[0282] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 115, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 116, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 117. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 118. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 119. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 281.

[0283] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 120, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 121, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 122, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 125, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 126, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 127.

[0284] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 120, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 121, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 122. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 123. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 124. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 282.

[0285] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 125, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 126, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 127. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 128. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 129. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 283.

[0286] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 130, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 131, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 132, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 135, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 136, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 137.

[0287] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 130, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 131, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 132. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 133. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 134. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 284.

[0288] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 135, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 136, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 137. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the beta chain comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 139. In some embodiments, the beta chain comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 285.

[0289] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 140, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 141, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 142, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 145, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 146, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 147.

[0290] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 140, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 141, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 142. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 143. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 144. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 286.

[0291] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 145, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 146, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 147. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 148. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 149. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 287.

[0292] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 150, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 151, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 152, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 155, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 156, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 157.

[0293] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of aa SEQ ID NO: 150, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 151, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 152. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 153. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 154. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 288.

[0294] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 155, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 156, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 157. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 158. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 159. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 289.

[0295] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 160, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 161, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 162, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 165, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 166, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 167.

[0296] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 160, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 161, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 162. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 163. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 164. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 290.

[0297] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 165, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 166, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 167. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 168. In some embodiments, the beta chain comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 169. In some embodiments, the beta chain comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 291.

[0298] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 170, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 171, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 172, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 175, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 176, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 177.

[0299] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 170, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 171, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 173. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 174. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 292.

[0300] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 175, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 176, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 177. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 178. In some embodiments, the beta chain comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 179. In some embodiments, the beta chain comprises an amino acid sequence at least 90%, 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 293.

[0301] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 180, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 181, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 182, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 185, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 186, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 187.

[0302] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 180, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 181, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 182. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 183. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 294. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 184.

[0303] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 185, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 186, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 187. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 188. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 295. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 189.

[0304] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 190, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 191, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 192, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 195, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 196, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 197.

[0305] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 190, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 191, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 192. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 193. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 296. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 194.

[0306] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 195, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 196, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 197. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 198. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 297. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 199.

[0307] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 200, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 201, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 202, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 205, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 206, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 207.

[0308] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 200, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 201, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 202. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 203. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 298. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 204.

[0309] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 205, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 206, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 207. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 208. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 299. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 209.

[0310] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 210, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 211, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 212, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 215, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 216, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 217.

[0311] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 210, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 211, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 212. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 213. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 300. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 214.

[0312] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 215, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 216, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 217. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 218. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 301. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 219.

[0313] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 220, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 221, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 222, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 225, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 226, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 227.

[0314] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 220, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 221, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 222. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 223. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 302. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 224.

[0315] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 225, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 226, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 227. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 228. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 303. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 229.

[0316] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 230, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 231, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 232, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 235, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 236, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 237.

[0317] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 230, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 231, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 232. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 233. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 304. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 234.

[0318] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 235, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 236, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 237. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 238. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 305. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 239.

[0319] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 240, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 241, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 242, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 245, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 246, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 247.

[0320] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 240, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 241, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 242. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 243. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 306. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 244.

[0321] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 245, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 246, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 247. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 248. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 307. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 249.

[0322] Some embodiments relate to engineered regulatory T cells comprising an exogenous TCR that binds to a proinsulin peptide complexed with an MHC molecule, wherein the TCR comprises an alpha chain and a beta chain, and wherein the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 250, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 251, a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 252, a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 255, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 256, and / or a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 257.

[0323] In some embodiments, the TCR comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 250, a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 251, and a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 252. In some embodiments, the alpha chain comprises an alpha variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 253. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 308. In some embodiments, the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 254.

[0324] In some embodiments, the TCR comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 255, a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO: 256, and a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO: 257. In some embodiments, the beta chain comprises a beta variable region comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 258. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 309. In some embodiments, the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 259.

[0325] group In some embodiments, the present disclosure provides cell populations (e.g., isolated cell populations) comprising regulatory T cells, including stable regulatory T cells transfected with an exogenous human TCR (e.g., a TCR listed in Table 1). Accordingly, in some embodiments, the present disclosure provides isolated cell populations comprising stable regulatory T cells comprising a hypomethylated TSDR at the endogenous FOXP3 locus. In some embodiments, the stable regulatory T cells further comprise an exogenous human TCR that specifically binds to a target peptide complexed with an MHC molecule.

[0326] An isolated cell population is a cell population that has been removed from a human body. Thus, it is considered "isolated" from the human body. The population of cells can be isolated (e.g., obtained) from a subject or a biological sample obtained from a subject, for example, using any known cell collection method, such as apheresis. The isolated cell population of the present disclosure can be subjected to the methods described herein to produce an isolated cell population with a higher number of regulatory T cells (e.g., stable regulatory T cells), for example, by apheresis relative to a population of cells obtained directly from a subject or a biological sample obtained from a subject, for example, using apheresis.

[0327] In some embodiments, the isolated cell population is CD25 + / high CD4 + CD127 - / lo In some embodiments, the isolated cell population comprises CD25 regulatory T cells. + / high CD4 + CD127 - / lo / FOXP3 + In some embodiments, the isolated cell population comprises CD25 regulatory T cells. + / high CD4 + CD127 - / lo / FOXP3 + / CD45RA + Includes regulatory T cells.

[0328] In some embodiments, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the cells of the isolated cell population are stable regulatory T cells that comprise a hypomethylated TSDR at the endogenous FOXP3 locus.

[0329] In some embodiments, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the cells in the isolated cell population are CD25 + / high CD4 + CD127 - / lo In some embodiments, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the cells in the isolated cell population are CD25 + / high CD4 + CD127 - / lo / FOXP3 + Regulatory T cells.

[0330] In some embodiments, at least 10%, at least 25%, at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the stable regulatory T cells of the population are CD45RA + In some embodiments, at least 10%, at least 25%, at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the stable regulatory T cells of the population are CD25 + / high CD4 + CD127 - / lo / CD45RA + In some embodiments, at least 10%, at least 25%, at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the stable regulatory T cells of the population are CD25 + / high CD4 + CD127 - / lo / FOXP3 + / CD45RA + is.

[0331] In some embodiments, the stable regulatory T cells of the cell population are not engineered at the endogenous FOXP3 locus. The FOXP3 locus is considered engineered if it has been edited (e.g., directly edited) using gene editing techniques, such as homology-directed repair (HDR)-based gene editing, or if the cells have been otherwise genetically modified to increase or stabilize expression from the FOXP3 locus relative to endogenous (naturally occurring) FOXP3 expression. In some embodiments, the endogenous FOXP3 in the regulatory T cells of the population has not been gene-edited. Thus, in some embodiments, fewer than 10% (less than) of the cells of the isolated cell population express FOXP3 protein from an engineered FOXP3 locus. In some embodiments, fewer than (less than) 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the cells of the isolated cell population express FOXP3 protein from the engineered FOXP3 locus. In some embodiments, fewer than (less than) 0.5% or less than 0.1% of the cells of the isolated cell population express FOXP3 protein from the engineered FOXP3 locus.

[0332] In some embodiments, the isolated cell population comprises at least 1 x 10 2 , at least 1 x 10 3 , at least 1 x 10 4 , at least 1 x 10 5 , at least 1 x 10 6 , at least 1 x 10 7 , at least 1 x 10 8 , at least 1 x 10 9 , or at least 1 × 10 10 In some embodiments, the isolated cell population comprises 1 x 10 stable regulatory T cells. 2 ~1×10 10 , 1×10 3 ~1×1010 , 1×10 4 ~1×10 10 , 1×10 5 ~1×10 10 , 1×10 6 ~1×10 10 , 1×10 7 ~1×10 10 , 1×10 8 ~1×10 10 , 1×10 5 ~1×10 9 , 1×10 6 ~1×10 8 , 1×10 7 ~1×10 10 , or 1 × 10 4 ~1×10 6 In some embodiments, the isolated cell population comprises 1 x 10 stable regulatory T cells. 6 ~1×10 10 Contains stable regulatory T cells.

[0333] In some embodiments, a subset of the isolated cell population is stable regulatory T cells (e.g., those with a hypomethylated TSDR at the endogenous FOXP3 locus and expressing CD4, CD25, and FOXP3). In some embodiments, the stable regulatory T cells within the isolated cell population maintain a hypomethylated TSDR at the endogenous FOXP3 locus over time, as described elsewhere herein.

[0334] In some embodiments, stable regulatory T cells within the isolated cell population retain markers of stability in the presence of pro-inflammatory conditions (e.g., in the presence of one or more pro-inflammatory cytokines). In some embodiments, stable regulatory T cells within the isolated cell population maintain a hypomethylated TSDR at the endogenous FOXP3 locus in the presence of pro-inflammatory conditions (e.g., in the presence of one or more pro-inflammatory cytokines). In some embodiments, stable regulatory T cells within the isolated cell population maintain a hypomethylated TSDR at the endogenous FOXP3 locus and express CD4, CD25, and FOXP3 in the presence of pro-inflammatory conditions (e.g., in the presence of one or more pro-inflammatory cytokines). In some embodiments, the stable regulatory T cells within the isolated cell population maintain a hypomethylated TSDR at the endogenous FOXP3 locus in the presence of a pro-inflammatory condition (e.g., in the presence of one or more pro-inflammatory cytokines) for at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 21, 22, 23, 24, or 25 days.

[0335] At least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% of the cells of the isolated cell population can be stable regulatory T cells. Thus, in some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus. In some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days (e.g., after being obtained from the subject or after transduction with five or fewer copies of a nucleic acid expressing a TCR, e.g., a nucleic acid expressing a TCR).

[0336] In some embodiments, at least 50% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 5 days. In some embodiments, at least 50% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 10 days. In some embodiments, at least 50% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 15 days. In some embodiments, at least 60% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 5 days. In some embodiments, at least 60% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 10 days. In some embodiments, at least 60% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 15 days. In some embodiments, at least 70% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 5 days. In some embodiments, at least 70% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 10 days. In some embodiments, at least 70% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 15 days. In some embodiments, at least 80% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 5 days. In some embodiments, at least 80% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 10 days. In some embodiments, at least 80% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 15 days. In some embodiments, at least 90% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 5 days.In some embodiments, at least 90% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 10 days, hi some embodiments, at least 90% of the cells of the isolated cell population comprise a hypomethylated TSDR at the FOXP3 locus for at least 15 days.

[0337] In some embodiments, the percentage of cells in the isolated cell population comprising stable regulatory T cells does not decrease by more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 percentage point during at least 5, 6, 7, 8, 9, 10, or 11 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in the isolated cell population comprising stable regulatory T cells increases by 1, 2, 3, 4, or 5 percentage points during at least 5, 6, 7, 8, 9, 10, or 11 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in the isolated cell population comprising stable regulatory T cells does not decrease by more than 10 percentage points during at least 5 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in an isolated cell population comprising stable regulatory T cells does not decrease by more than 10 percentage points during at least 10 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in an isolated cell population comprising stable regulatory T cells does not decrease by more than 10 percentage points during at least 15 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in an isolated cell population comprising stable regulatory T cells does not decrease by more than 5 percentage points during at least 5 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in an isolated cell population comprising stable regulatory T cells does not decrease by more than 5 percentage points during at least 10 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in the isolated cell population comprising stable regulatory T cells does not decrease by more than 5 percentage points during at least 15 days of expansion following transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR.In some embodiments, the percentage of cells in an isolated cell population comprising stable regulatory T cells does not decrease by more than 1 percentage point during at least 5 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in an isolated cell population comprising stable regulatory T cells does not decrease by more than 1 percentage point during at least 10 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR. In some embodiments, the percentage of cells in an isolated cell population comprising stable regulatory T cells does not decrease by more than 1 percentage point during at least 15 days of expansion after transduction of the stable regulatory T cells with a nucleic acid expressing an exogenous human TCR.

[0338] The percentage of stable regulatory T cells relative to total cells in an isolated cell population comprising regulatory T cells can be assessed about 1, about 6, about 12, about 24, about 36, about 48, about 72, about 96, or about 120 hours after transduction of the cells with a nucleic acid expressing an exogenous human TCR. The percentage of stable regulatory T cells relative to total cells in an isolated cell population comprising regulatory T cells can be assessed 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days after transduction of the cells with a nucleic acid expressing an exogenous human TCR. The percentage of stable regulatory T cells relative to total cells in an isolated cell population comprising regulatory T cells can be assessed about 1-21, 1-7, 4-14, 4-7, 7-10, 7-14, 10-21, or 14-21 days after transduction of the cells with a nucleic acid expressing an exogenous human TCR.

[0339] In some embodiments, at least 10%, at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of the cells of the isolated cell population comprise an unmodified endogenous FOXP3 locus. In some embodiments, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2%, or less than 1% of the cells of the isolated cell population comprise a modified endogenous FOXP3 locus. An unmodified endogenous FOXP3 locus is a naturally occurring FOXP3 locus, which has not been edited, for example, using gene editing or transgenic techniques.

[0340] In some embodiments, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of the cells of the isolated cell population express physiological levels of functional FOXP3 protein, where physiological levels of functional FOXP3 protein are the levels of protein expressed from an unmodified (naturally occurring) FOXP3 locus.

[0341] In some embodiments, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of the cells of the isolated cell population are free of an exogenous, e.g., ectopic, FOXP3 gene or exogenous, e.g., ectopic FOXP3 protein. In some embodiments, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of the cells of the isolated cell population are free of ectopic expression of FOXP3. Ectopic expression of FOXP3 refers to expression that does not naturally occur in a cell. Thus, ectopic expression can result, for example, from expression of an exogenous transgene or from expression from an endogenous FOXP3 gene that has been modified at the FOXP3 locus, e.g., in the promoter region of the FOXP3 gene.

[0342] In some embodiments, less than 40%, less than 30%, less than 20%, less than 10%, less than 5%, less than 2%, or less than 1% of the cells of the isolated cell population comprise an exogenous, e.g., ectopic, FOXP3 gene or exogenous, e.g., ectopic FOXP3 protein, which has been introduced into the cells by, for example, transfection, transduction, or electroporation.

[0343] In some embodiments, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the regulatory T cells of the isolated cell population of regulatory T cells express an exogenous TCR (e.g., after transduction of the isolated cell population with a nucleic acid encoding an exogenous TCR). In some embodiments, between 10% and 60% or between 20% and 50% of the regulatory T cells of the isolated cell population of regulatory T cells express an exogenous TCR. In some embodiments, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the stable regulatory T cells of the isolated cell population of regulatory T cells express an exogenous TCR (e.g., after transduction of the isolated cell population with an exogenous TCR). In some embodiments, between 10% and 60%, or between 20% and 50% of the stable regulatory T cells of the isolated cell population of regulatory T cells express an exogenous TCR.

[0344] In some embodiments, the TSDR at the endogenous FOXP3 locus of stable regulatory T cells of the isolated cell population of regulatory T cells remains hypomethylated until administration of the isolated cell population to a subject. In some embodiments, the TSDR at the endogenous FOXP3 locus of stable regulatory T cells of the isolated cell population of regulatory T cells remains hypomethylated after cryopreservation freeze-thaw cycles.

[0345] In some embodiments, the regulatory T cells (e.g., stable regulatory T cells) of the isolated cell population exhibit one or more cellular functions associated with regulatory T cells when activated by binding to pMHC. Non-limiting examples of such cellular functions include cytokine secretion activity, expression of specific activation markers, and suppressive activity. Cytokine secretion activity simply refers to the secretion of certain anti-inflammatory cytokines, such as IL-10, IL-4, IL-6, IL-11, and IL-13. Activation markers include, but are not limited to, CD69, 4-1BB, CD25, CD71, or CTLA-4. Regulatory T cells express one or more of these markers, for example, upon contact with a target peptide complexed with an MHC molecule.

[0346] Suppressive activity refers to the activity of non-regulatory T cells (e.g., CD8 + T cells and CD4 +Regulatory T cells partially suppress the activation, proliferation, and cytokine production of conventional T cells (e.g., T cells that inhibit immune system overactivity). Regulatory T cells exhibit suppressive activity when contacted with a target peptide complexed with an MHC molecule that binds to an exogenous TCR expressed by the regulatory T cells. In some embodiments, the target peptide is presented by a cell expressing the MHC molecule. For example, in some embodiments, regulatory T cells suppress the activation, proliferation, and cytokine production of conventional T cells that have specificity for a common target peptide complexed with an MHC molecule, e.g., a common target peptide presented by an antigen-presenting cell (APC). In some embodiments, regulatory T cells suppress the proliferation and growth of non-regulatory T cells by at least 25%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% relative to a control (e.g., non-regulatory T cells in the absence of the regulatory T cells).

[0347] In some embodiments, regulatory T cells suppress interferon (IFN)-gamma production from conventional T cells by at least 25%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% relative to a control (e.g., conventional T cells in the absence of regulatory T cells). In some embodiments, regulatory T cells suppress IFN-gamma production from conventional T cells by 50% to 99%, 75% to 99%, or 80% to 100% relative to a control (e.g., when present in a population comprising a 1:1 to 1:8 ratio of regulatory T cells compared to conventional T cells). In some embodiments, regulatory T cells suppress CD71 production from conventional T cells by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, or at least 70% relative to a control (e.g., conventional T cells in the absence of regulatory T cells). In some embodiments, the regulatory T cells suppress CD71 production by conventional T cells by 20% to 90% or 30% to 80% relative to a control (e.g., when present in a population comprising a 1:1 to 1:8 ratio of regulatory T cells compared to conventional T cells). In some embodiments, the regulatory T cells suppress CD25 production from conventional T cells by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, or at least 70% relative to a control (e.g., conventional T cells in the absence of regulatory T cells). In some embodiments, the regulatory T cells suppress CD25 production by conventional T cells by 40% to 70% relative to a control (e.g., when present in a population comprising a 1:1 to 1:8 ratio of regulatory T cells compared to conventional T cells).

[0348] In some embodiments, regulatory T cells exhibit cytokine secretion activity (e.g., secretion of IL-10) upon contact with a target peptide complexed with an MHC molecule that binds to an exogenous TCR expressed by the regulatory T cells, e.g., a target peptide presented by an APC. In some embodiments, regulatory T cells exhibit expression of activation markers upon contact with a target peptide complexed with an MHC molecule that binds to an exogenous TCR expressed by the regulatory T cells. For example, in some embodiments, regulatory T cells exhibit expression of CD69, 4-1BB, CD25, CD71, and / or CTLA-4 upon contact with a target peptide complexed with an MHC molecule that binds to an exogenous TCR expressed by the regulatory T cells. In some embodiments, regulatory T cells exhibit suppressive activity upon contact with a target peptide complexed with an MHC molecule that binds to an exogenous TCR expressed by the regulatory T cells. For example, in some embodiments, regulatory T cells suppress the activation of conventional T cells with specificity for a common target peptide complexed with an MHC molecule.

[0349] The cellular function of stable regulatory T cells can be assessed about 1, about 6, about 12, about 24, about 36, about 48, about 72, about 96, or about 120 hours after transduction of the cells with a nucleic acid expressing an exogenous human TCR. The cellular function of stable regulatory T cells can be assessed 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days after transduction of the cells with a nucleic acid expressing an exogenous human TCR. The cellular function of stable regulatory T cells can be assessed about 1-21, 1-7, 4-14, 4-7, 7-10, 7-14, 10-21, or 14-21 days after transduction of the cells with a nucleic acid expressing an exogenous human TCR.

[0350] In some embodiments, transduced regulatory T cells (e.g., isolated populations of transduced regulatory T cells) retain their cellular function (e.g., the ability to be activated) after cryopreservation freeze-thaw cycling. In some embodiments, transduced regulatory T cells may be activated and / or expanded after cryopreservation freeze-thaw cycling. In some embodiments, regulatory T cells may exhibit cytokine secretion activity, expression of specific activation markers, and / or suppressive activity after cryopreservation freeze-thaw cycling.

[0351] In some embodiments, at least 5%, at least 10%, at least 25%, at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the population of regulatory T cells are CD25 T cells prior to activation and / or transduction with an exogenous human T cell receptor-expressing nucleic acid. + / high CD4 + CD127 - / lo In some embodiments, at least 5%, at least 10%, at least 25%, at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the population of regulatory T cells are CD25 T cells prior to activation and / or transduction with an exogenous human T cell receptor-expressing nucleic acid. + / high CD4 + CD127 - / lo CD45RA + is.

[0352] The isolated cell population containing regulatory T cells may contain a small amount of non-regulatory T cells (e.g., conventional T cells). Non-regulatory T cells include NK T cells, B cells, and CD8 T cells derived from peripheral blood and lymph nodes. + T cells, neutrophils, eosinophils, CD14 + Cellular, or conventional (CD4 +) T cells. In some embodiments, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, or less than 0.01% of the cells in an isolated cell population comprising regulatory T cells are non-regulatory T cells. In some embodiments, about 0.01% to about 0.1%, about 0.1% to about 0.5%, about 0.5% to about 1%, about 0.5% to about 10%, about 2% to about 5%, or about 5% to about 10% of the cells in an isolated cell population comprising regulatory T cells are non-regulatory T cells. In some embodiments, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, or less than 0.01% of the cells of an isolated cell population comprising regulatory T cells are non-regulatory T cells that comprise an exogenous human TCR (e.g., an engineered human TCR of the present disclosure). In some embodiments, no more than 10% of the cells of an isolated cell population comprising regulatory T cells are non-regulatory T cells that comprise an exogenous human TCR.

[0353] Conventional T cells generally produce IL-2 and other interleukin factors. In some embodiments, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, or less than 0.01% of the cells in an isolated cell population comprising regulatory T cells are conventional T cells. In some embodiments, about 0.01%, about 0.1%, about 0.1% to about 0.5%, about 0.5% to about 1%, about 0.5% to about 10%, about 2% to about 5%, or about 5% to about 10% of the cells in an isolated cell population comprising regulatory T cells are conventional T cells. In some embodiments, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, or less than 0.01% of the cells of an isolated cell population comprising regulatory T cells are conventional T cells comprising an exogenous human TCR (e.g., an engineered human TCR of the present disclosure). In some embodiments, an isolated cell population comprising regulatory T cells comprises an undetectable amount of conventional T cells. In some embodiments, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, or less than 0.01% of the cells of an isolated cell population comprising regulatory T cells are conventional CD4 T cells. + In some embodiments, 20% or less of the cells in the isolated cell population comprising regulatory T cells are conventional CD4 T cells. + In some embodiments, the isolated cell population comprising regulatory T cells contains undetectable amounts of conventional CD4 T cells. + A cell type is considered to be undetectable if the presence of the cell in a sample is not detectable by flow cytometry, for example, based on one or more cell-specific biomarkers.

[0354] In some embodiments, the isolated cell population comprising regulatory T cells comprises an undetectable amount of B cells. In some embodiments, the isolated cell population comprising regulatory T cells comprises an undetectable amount of NK T cells. In some embodiments, the isolated cell population comprising regulatory T cells comprises an undetectable amount of CD14 +In some embodiments, the isolated cell population comprising regulatory T cells comprises an undetectable number of eosinophils. In some embodiments, the isolated cell population comprising regulatory T cells comprises an undetectable number of neutrophils.

[0355] In some embodiments, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, or less than 0.01% of the cells of the isolated cell population comprising regulatory T cells are CD8 + In some embodiments, 5% or less of the cells in the isolated cell population are CD8 T cells. + In some embodiments, the isolated cell population comprising regulatory T cells contains undetectable amounts of CD8 + Contains T cells.

[0356] The percentage of conventional T cells or other non-regulatory T cells relative to total cells (or relative to stable regulatory T cells) can be assessed about 1, about 6, about 12, about 24, about 36, about 48, about 72, about 96, or about 120 hours after transduction of the cells with a nucleic acid expressing an exogenous human TCR. The percentage of conventional T cells or other non-regulatory T cells relative to total cells (or relative to stable regulatory T cells) can be assessed 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days after transduction of the cells with a nucleic acid expressing an exogenous human TCR. The percentage of conventional T cells or other non-regulatory T cells relative to total cells (or relative to stable regulatory T cells) can be assessed at about 1-21, 1-7, 4-14, 4-7, 7-10, 7-14, 10-21, or 14-21 days after transduction of the cells with a nucleic acid expressing an exogenous human TCR.

[0357] In some embodiments, the ratio of regulatory T cells to conventional T cells in the isolated population of cells comprising regulatory T cells is at least 5:1, at least 10:1, at least 15:1, at least 20:1, at least 25:1, at least 30:1, at least 35:1, at least 40:1, at least 45:1, at least 50:1, at least 60:1, at least 70:1, at least 80:1, at least 90:1, or at least 100:1.

[0358] Pharmaceutical Composition In some embodiments, the present disclosure provides a pharmaceutical composition comprising any one of the TCRs described herein (e.g., a TCR described in Table 1). In some embodiments, the present disclosure provides a pharmaceutical composition comprising an isolated cell population of regulatory T cells (e.g., stable regulatory T cells) described herein. In some embodiments, the pharmaceutical composition comprises a cell population of regulatory T cells (e.g., stable regulatory T cells) described herein and a pharmaceutically acceptable excipient.

[0359] As used herein, a pharmaceutically acceptable excipient may also be referred to as a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, or a pharmaceutically acceptable adjuvant. The formulation of pharmaceutically acceptable excipient and carrier solutions is well known to those skilled in the art, as is the development of appropriate dosing and treatment regimens for use with the particular compositions described herein in various treatment regimens.

[0360] Pharmaceutical compositions should typically be sterile and stable under the conditions of manufacture and storage. Sterile injectable formulations may be prepared using non-toxic parenterally acceptable diluents or solvents. Pharmaceutical compositions for use according to the present invention may contain pharmaceutically acceptable dispersing agents, wetting agents, suspending agents, isotonic agents, coatings, antibacterial and antifungal agents, carriers, excipients, salts, or stabilizers, which are non-toxic to subjects at the dosages and concentrations used. In some embodiments, the pharmaceutical composition may contain an organic solvent, such as, but not limited to, methyl acetate, dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), dimethoxyethane (DME), and dimethylacetamide (including mixtures or combinations thereof).

[0361] The pharmaceutical composition may comprise an effective amount of stable regulatory T cells sufficient to induce a desired biological response. For example, in some embodiments, the effective amount of stable regulatory T cells described herein may refer to a number of cells sufficient to ameliorate symptoms associated with type 1 diabetes. As will be understood by those skilled in the art, the effecti...

Claims

1. An engineered T cell receptor that specifically binds to the preproinsulin 73-90 peptide complexed with a major histocompatibility complex (MHC) molecule.

2. 2. The engineered T cell receptor of claim 1, wherein the engineered T cell receptor comprises an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of any one of SEQ ID NOs: 3, 33, 53, and 73, and / or the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of any one of SEQ ID NOs: 8, 38, 58, and 78.

3. 3. The engineered T cell receptor of claim 2, wherein the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO:3, and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:

8.

4. 4. The engineered T cell receptor of claim 3, wherein the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 1 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 2, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 6 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO:

7.

5. 5. The engineered T cell receptor of claim 3 or 4, wherein the alpha chain comprises an alpha variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO:4, and the beta chain comprises a beta variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO:

9.

6. 6. The engineered T cell receptor of any one of claims 3 to 5, wherein the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 5 or 260, and the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 10 or 261.

7. 7. The engineered T cell receptor of claim 6, comprising the sequence of SEQ ID NO: 310 or 311.

8. 3. The engineered T cell receptor of claim 2, wherein the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 33, and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:

38.

9. 9. The engineered T cell receptor of claim 8, wherein the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 31 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 32, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 36 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO:

37.

10. 10. The engineered T cell receptor of claim 8 or 9, wherein the alpha chain comprises an alpha variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 34, and the beta chain comprises a beta variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO:

39.

11. 11. The engineered T cell receptor of any one of claims 8-10, wherein the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 35 or 266, and the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 40 or 267.

12. 12. The engineered T cell receptor of claim 11, comprising the sequence of SEQ ID NO: 312 or 313.

13. 3. The engineered T cell receptor of claim 2, wherein the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 53, and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:

58.

14. 14. The engineered T cell receptor of claim 13, wherein the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO:51 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO:52, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO:56 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO:

57.

15. 15. The engineered T cell receptor of claim 13 or 14, wherein the alpha chain comprises an alpha variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 54, and the beta chain comprises a beta variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO:

59.

16. 16. The engineered T cell receptor of any one of claims 13-15, wherein the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 55 or 270, and the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 60 or 271.

17. 17. The engineered T cell receptor of claim 16, comprising the sequence of SEQ ID NO: 314 or 315.

18. 3. The engineered T cell receptor of claim 2, wherein the alpha chain comprises a CDR3 alpha sequence comprising the amino acid sequence of SEQ ID NO: 73 and the beta chain comprises a CDR3 beta sequence comprising the amino acid sequence of SEQ ID NO:

78.

19. 19. The engineered T cell receptor of claim 18, wherein the alpha chain comprises a CDR1 alpha sequence comprising the amino acid sequence of SEQ ID NO: 71 and a CDR2 alpha sequence comprising the amino acid sequence of SEQ ID NO: 72, and the beta chain comprises a CDR1 beta sequence comprising the amino acid sequence of SEQ ID NO: 76 and a CDR2 beta sequence comprising the amino acid sequence of SEQ ID NO:

77.

20. 20. The engineered T cell receptor of claim 18 or 19, wherein the alpha chain comprises an alpha variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 74, and the beta chain comprises a beta variable domain comprising an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO:

79.

21. 21. The engineered T cell receptor of any one of claims 18-20, wherein the alpha chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 75 or 274, and the beta chain comprises an amino acid sequence having at least 90%, 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 80 or 275.

22. 22. The engineered T cell receptor of claim 21, comprising the sequence of SEQ ID NO: 316 or 317.

23. 23. The engineered T cell receptor of any one of claims 1 to 22, which is encoded as a single polypeptide.

24. 24. The engineered T cell receptor of any one of claims 2 to 23, further comprising a self-cleaving peptide sequence located between the alpha chain and the beta chain.

25. 25. The engineered T cell receptor of claim 24, wherein the self-cleaving peptide sequence is a 2A peptide sequence, optionally wherein the 2A peptide sequence is a P2A, E2A, F2A, or T2A peptide sequence.

26. 26. The engineered T cell receptor of any one of claims 1 to 25, wherein the preproinsulin 73-90 peptide comprises the amino acid sequence GAGSLQPLALEGSLQKRG (SEQ ID NO: 109).

27. 27. The engineered T cell receptor of any one of claims 1 to 26, wherein the MHC molecule comprises an HLA-DRB1*04:01 molecule.

28. 28. An engineered nucleic acid encoding an engineered T cell receptor according to any one of claims 1 to 27.

29. 29. The engineered nucleic acid of Claim 28, wherein the engineered nucleic acid is a viral vector, optionally a lentiviral vector.

30. 30. The engineered nucleic acid of claim 28 or 29, wherein the engineered nucleic acid comprises an EF-1 alpha promoter or an MND promoter operably linked to a sequence encoding the engineered T cell receptor.

31. 31. The engineered nucleic acid of any one of claims 28-30, wherein the engineered nucleic acid further comprises an enhancer element, optionally an optimized post-transcriptional regulatory element (oPRE) or a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), and optionally a WPRE-mut6.

32. Regulatory T cells comprising an engineered T cell receptor that specifically binds to preproinsulin 73-90 peptide complexed with a major histocompatibility complex (MHC) molecule.

33. The regulatory T cell of claim 32, wherein the engineered T cell receptor is an engineered T cell receptor of any one of claims 1 to 27.

34. The regulatory T cell of claim 32, wherein the engineered T cell receptor expresses an engineered nucleic acid of any one of claims 28 to 31.

35. The regulatory T cell of any one of claims 32 to 34, wherein the regulatory T cell is phenotypically stable.

36. 36. The regulatory T cell of any one of claims 32-35, wherein the regulatory T cell has not been gene edited at the endogenous FOXP3 locus.

37. 37. A method of treating type 1 diabetes in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising regulatory T cells of any one of claims 32-36.

38. 38. The method of claim 37, wherein the regulatory T cells are autologous to the subject.

39. 39. The method of claim 37 or 38, wherein the subject has type 1 diabetes, optionally stage 3 type 1 diabetes.

40. The method of any one of claims 37 to 39, wherein the subject is an HLA-DRB1*04:01 positive subject.

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