T cell receptor targeting the minor histocompatibility antigen HA-2

Abstract

Provided herein are T cell receptors (TCRs) or antigen-binding fragments thereof, such as those that recognize or bind to relatively hematopoietically restricted minor histocompatibility antigens, e.g., HA-2. In particular, the disclosure relates to TCRs that bind to or recognize specific HA-2 peptides in the context of major histocompatibility complex (MHC) molecules. The disclosure further relates to nucleic acids encoding such TCRs, engineered cells containing such TCRs, methods for isolating such TCRs, and their uses, e.g., in cell therapy.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Patent Application No. 63 / 385,357, filed November 29, 2022, and U.S. Provisional Patent Application No. 63 / 496,766, filed April 18, 2023, each of which is incorporated herein by reference.

[0002] Sequence Listing The sequence listing set forth in the file BSB-0006WO001_SeqListing.xml is 385 kilobytes in size, was created on November 28, 2023, and is incorporated herein by reference. [Background technology]

[0003] Allogeneic hematopoietic stem cell transplantation (HSCT) (HLA-matched or HLA-haploidentical) can be used to treat diseases or conditions such as hematologic malignancies and other non-malignant conditions. Some subjects may relapse after alloSCT. Improved therapies are needed to achieve optimal treatment outcomes. Embodiments are provided that meet such needs. Summary of the Invention

[0004] T cell receptors (TCRs), or antigen-binding fragments thereof, are described that recognize or bind to relatively hematopoietically restricted minor histocompatibility antigens, such as HA-2. In particular, the TCRs bind to or recognize specific HA-2 peptides in the context of major histocompatibility complex (MHC) molecules. The present disclosure further relates to nucleic acids encoding such TCRs, engineered cells containing such TCRs, methods for isolating such TCRs, and their uses, e.g., in cell therapy.

[0005] a TCR or antigen-binding fragment thereof, comprising an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises a complementarity determining region 3 (CDR-3) comprising SEQ ID NO: 13, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 21; or (b) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 31, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 39; or (c) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 49, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 57; or (d) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 67, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 75; or (e) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 85, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 93; or (f) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 103, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 111; or (g) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 121, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 129; or (h) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 139, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 147; or (i) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 157, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 165; or (j) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 175, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 183; or (k) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 193, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 201; or (l) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 211, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 219; or (m) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 229, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 237; or (n) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 247, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 255; or (o) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 265, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 273; or (p) Provided herein is a TCR or antigen-binding fragment thereof, wherein the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 283, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 291.

[0006] Also, a TCR or an antigen-binding fragment thereof, comprising an alpha chain comprising a Vα region and a beta chain comprising a Vβ region, or a gamma chain comprising a Vγ region and a delta chain comprising a Vδ region, (a) the Vα or Vγ region comprises CDR-1 comprising SEQ ID NO: 11, CDR-2 comprising SEQ ID NO: 12, and CDR-3 comprising SEQ ID NO: 13, and the Vβ or Vδ region comprises CDR-1 comprising SEQ ID NO: 19, CDR-2 comprising SEQ ID NO: 20, and CDR-3 comprising SEQ ID NO: 21; or (b) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 29, a CDR-2 comprising SEQ ID NO: 30, and a CDR-3 comprising SEQ ID NO: 31, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 37, a CDR-2 comprising SEQ ID NO: 38, and a CDR-3 comprising SEQ ID NO: 38; or (c) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 47, a CDR-2 comprising SEQ ID NO: 48, and a CDR-3 comprising SEQ ID NO: 49, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 55, a CDR-2 comprising SEQ ID NO: 56, and a CDR-3 comprising SEQ ID NO: 57; or (d) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 65, a CDR-2 comprising SEQ ID NO: 66, and a CDR-3 comprising SEQ ID NO: 67, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 73, a CDR-2 comprising SEQ ID NO: 74, and a CDR-3 comprising SEQ ID NO: 75; or (e) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 83, a CDR-2 comprising SEQ ID NO: 84, and a CDR-3 comprising SEQ ID NO: 85, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 91, a CDR-2 comprising SEQ ID NO: 92, and a CDR-3 comprising SEQ ID NO: 93; or (f) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 101, a CDR-2 comprising SEQ ID NO: 102, and a CDR-3 comprising SEQ ID NO: 103, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 109, a CDR-2 comprising SEQ ID NO: 110, and a CDR-3 comprising SEQ ID NO: 111; or (g) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 119, a CDR-2 comprising SEQ ID NO: 120, and a CDR-3 comprising SEQ ID NO: 121, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 127, a CDR-2 comprising SEQ ID NO: 128, and a CDR-3 comprising SEQ ID NO: 129; or (h) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 137, a CDR-2 comprising SEQ ID NO: 138, and a CDR-3 comprising SEQ ID NO: 139, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 145, a CDR-2 comprising SEQ ID NO: 146, and a CDR-3 comprising SEQ ID NO: 147; or (i) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 155, a CDR-2 comprising SEQ ID NO: 156, and a CDR-3 comprising SEQ ID NO: 157, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 163, a CDR-2 comprising SEQ ID NO: 164, and a CDR-3 comprising SEQ ID NO: 165; or (j) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 173, a CDR-2 comprising SEQ ID NO: 174, and a CDR-3 comprising SEQ ID NO: 175, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 181, a CDR-2 comprising SEQ ID NO: 182, and a CDR-3 comprising SEQ ID NO: 183; or (k) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 191, a CDR-2 comprising SEQ ID NO: 192, and a CDR-3 comprising SEQ ID NO: 193, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 199, a CDR-2 comprising SEQ ID NO: 200, and a CDR-3 comprising SEQ ID NO: 201; or (l) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 209, a CDR-2 comprising SEQ ID NO: 210, and a CDR-3 comprising SEQ ID NO: 211, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 217, a CDR-2 comprising SEQ ID NO: 218, and a CDR-3 comprising SEQ ID NO: 219; or (m) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 227, a CDR-2 comprising SEQ ID NO: 228, and a CDR-3 comprising SEQ ID NO: 229, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 235, a CDR-2 comprising SEQ ID NO: 236, and a CDR-3 comprising SEQ ID NO: 237; or (n) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 245, a CDR-2 comprising SEQ ID NO: 246, and a CDR-3 comprising SEQ ID NO: 247, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 253, a CDR-2 comprising SEQ ID NO: 254, and a CDR-3 comprising SEQ ID NO: 255; or (o) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 263, a CDR-2 comprising SEQ ID NO: 264, and a CDR-3 comprising SEQ ID NO: 265, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 271, a CDR-2 comprising SEQ ID NO: 272, and a CDR-3 comprising SEQ ID NO: 273; or (p) Provided herein is a TCR or antigen-binding fragment thereof, wherein the Vα or Vγ region comprises CDR-1 comprising SEQ ID NO: 281, CDR-2 comprising SEQ ID NO: 282, and CDR-3 comprising SEQ ID NO: 283, and the Vβ or Vδ region comprises CDR-1 comprising SEQ ID NO: 289, CDR-2 comprising SEQ ID NO: 290, and CDR-3 comprising SEQ ID NO: 291.

[0007] Also, a TCR or an antigen-binding fragment thereof, comprising an alpha chain comprising a Vα region and a beta chain comprising a Vβ region, or a gamma chain comprising a Vγ region and a delta chain comprising a Vδ region, (a) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 14, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 22; or (b) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 32, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 40; or (c) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 50, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 58; or (d) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 68, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 76; or (e) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 86, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 94; or (f) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 104, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 112; or (g) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 122, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 130; or (h) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 140, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 148; or (i) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 158, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 166; or (j) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 176, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 184; or (k) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 194, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 202; or (l) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 212, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 220; or (m) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 230, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 238; or (n) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 248, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 256; or (o) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 266, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 274; or (p) Provided herein is a TCR or antigen-binding fragment thereof, wherein the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 284, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 292.

[0008] In some embodiments, (a) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 14, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 22; or (b) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 32, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 40; or (c) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 50, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 58; or (d) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 68, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 76; or (e) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 86, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 94; or (f) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 104, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 112; or (g) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 122, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 130; or (h) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 140, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 148; or (i) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 158, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 166; or (j) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 176, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 184; or (k) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 194, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 202; or (l) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 212, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 220; or (m) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 230, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 238; or (n) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 248, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 256; or (o) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 266, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 274; or (p) The Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 284, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 292.

[0009] Also, a TCR or an antigen-binding fragment thereof, comprising an alpha chain comprising a Vα region and a beta chain comprising a Vβ region, or a gamma chain comprising a Vγ region and a delta chain comprising a Vδ region, (a) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 14, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 22; or (b) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 32, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 40; or (c) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 50, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 58; or (d) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 68, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 76; or (e) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 86, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 94; or (f) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 104, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 112; or (g) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 122, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 130; or (h) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 140, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 148; or (i) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 158, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 166; or (j) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 176, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 184; or (k) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 194, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 202; or (l) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 212, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 220; or (m) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 230, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 238; or (n) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 248, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 256; or (o) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 266, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 274; or (p) A TCR or antigen-binding fragment thereof is provided, wherein the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 284, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 292.

[0010] Also, a TCR or an antigen-binding fragment thereof, comprising an alpha chain comprising a Vα region and a beta chain comprising a Vβ region, or a gamma chain comprising a Vγ region and a delta chain comprising a Vδ region, (a) the Vα or Vγ region comprises SEQ ID NO: 14, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 22, or a sequence having at least 90% sequence identity thereto; or (b) the Vα or Vγ region comprises SEQ ID NO: 32, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 40, or a sequence having at least 90% sequence identity thereto; or (c) the Vα or Vγ region comprises SEQ ID NO: 50, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 58, or a sequence having at least 90% sequence identity thereto; or (d) the Vα or Vγ region comprises SEQ ID NO: 68, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 76, or a sequence having at least 90% sequence identity thereto; or (e) the Vα or Vγ region comprises SEQ ID NO: 86, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 94, or a sequence having at least 90% sequence identity thereto; or (f) the Vα or Vγ region comprises SEQ ID NO: 104, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 112, or a sequence having at least 90% sequence identity thereto; or (g) the Vα or Vγ region comprises SEQ ID NO: 122, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 130, or a sequence having at least 90% sequence identity thereto; or (h) the Vα or Vγ region comprises SEQ ID NO: 140, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 148, or a sequence having at least 90% sequence identity thereto; or (i) the Vα or Vγ region comprises SEQ ID NO: 158, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 166, or a sequence having at least 90% sequence identity thereto; or (j) the Vα or Vγ region comprises SEQ ID NO: 176, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 184, or a sequence having at least 90% sequence identity thereto; or (k) the Vα or Vγ region comprises SEQ ID NO: 194, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 202, or a sequence having at least 90% sequence identity thereto; or (l) the Vα or Vγ region comprises SEQ ID NO: 212, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 220, or a sequence having at least 90% sequence identity thereto; or (m) the Vα or Vγ region comprises SEQ ID NO: 230, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 238, or a sequence having at least 90% sequence identity thereto; or (n) the Vα or Vγ region comprises SEQ ID NO: 248, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 256, or a sequence having at least 90% sequence identity thereto; or (o) the Vα or Vγ region comprises SEQ ID NO: 266, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 274, or a sequence having at least 90% sequence identity thereto; or (p) There is provided a TCR or antigen-binding fragment thereof, wherein the Vα or Vγ region comprises SEQ ID NO: 284, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 292, or a sequence having at least 90% sequence identity thereto.

[0011] In some embodiments, (a) the Vα or Vγ region comprises SEQ ID NO: 14 and the Vβ or Vδ region comprises SEQ ID NO: 22; or (b) the Vα or Vγ region comprises SEQ ID NO: 32 and the Vβ or Vδ region comprises SEQ ID NO: 40; or (c) the Vα or Vγ region comprises SEQ ID NO: 50 and the Vβ or Vδ region comprises SEQ ID NO: 58; or (d) the Vα or Vγ region comprises SEQ ID NO: 68 and the Vβ or Vδ region comprises SEQ ID NO: 76; or (e) the Vα or Vγ region comprises SEQ ID NO: 86 and the Vβ or Vδ region comprises SEQ ID NO: 94; or (f) the Vα or Vγ region comprises SEQ ID NO: 104 and the Vβ or Vδ region comprises SEQ ID NO: 112; or (g) the Vα or Vγ region comprises SEQ ID NO: 122 and the Vβ or Vδ region comprises SEQ ID NO: 130; or (h) the Vα or Vγ region comprises SEQ ID NO: 140 and the Vβ or Vδ region comprises SEQ ID NO: 148; or (i) the Vα or Vγ region comprises SEQ ID NO: 158 and the Vβ or Vδ region comprises SEQ ID NO: 166; or (j) the Vα or Vγ region comprises SEQ ID NO: 176 and the Vβ or Vδ region comprises SEQ ID NO: 184; or (k) the Vα or Vγ region comprises SEQ ID NO: 194 and the Vβ or Vδ region comprises SEQ ID NO: 202; or (l) the Vα or Vγ region comprises SEQ ID NO: 212 and the Vβ or Vδ region comprises SEQ ID NO: 220; or (m) the Vα or Vγ region comprises SEQ ID NO: 230 and the Vβ or Vδ region comprises SEQ ID NO: 238; or (n) the Vα or Vγ region comprises SEQ ID NO: 248 and the Vβ or Vδ region comprises SEQ ID NO: 256; or (o) the Vα or Vγ region comprises SEQ ID NO: 266 and the Vβ or Vδ region comprises SEQ ID NO: 274; or (p) the Vα or Vγ region comprises SEQ ID NO: 284, and the Vβ or Vδ region comprises SEQ ID NO: 292.

[0012] In some embodiments, the alpha chain further comprises an alpha constant (Cα) region and the beta chain further comprises a beta constant (Cβ) region, or the gamma chain further comprises a gamma constant (Cγ) region and the delta chain further comprises a delta constant (Cδ) region. In some embodiments, Cα comprises SEQ ID NO: 3 or 5, and Cβ comprises SEQ ID NO: 7 or 9.

[0013] In some embodiments, (a) the alpha or gamma chain comprises SEQ ID NO: 17 and the beta or delta chain comprises SEQ ID NO: 25; or (b) the alpha or gamma chain comprises SEQ ID NO: 35 and the beta or delta chain comprises SEQ ID NO: 43; or (c) the alpha or gamma chain comprises SEQ ID NO: 53 and the beta or delta chain comprises SEQ ID NO: 61; or (d) the alpha or gamma chain comprises SEQ ID NO: 71 and the beta or delta chain comprises SEQ ID NO: 79; or (e) the alpha or gamma chain comprises SEQ ID NO: 89 and the beta or delta chain comprises SEQ ID NO: 97; or (f) the alpha or gamma chain comprises SEQ ID NO: 107 and the beta or delta chain comprises SEQ ID NO: 115; or (g) the alpha or gamma chain comprises SEQ ID NO: 125 and the beta or delta chain comprises SEQ ID NO: 133; or (h) the alpha or gamma chain comprises SEQ ID NO: 143 and the beta or delta chain comprises SEQ ID NO: 151; or (i) the alpha or gamma chain comprises SEQ ID NO: 161 and the beta or delta chain comprises SEQ ID NO: 169; or (j) the alpha or gamma chain comprises SEQ ID NO: 179 and the beta or delta chain comprises SEQ ID NO: 187; or (k) the alpha or gamma chain comprises SEQ ID NO: 197 and the beta or delta chain comprises SEQ ID NO: 205; or (l) the alpha or gamma chain comprises SEQ ID NO: 215 and the beta or delta chain comprises SEQ ID NO: 223; or (m) the alpha or gamma chain comprises SEQ ID NO: 233 and the beta or delta chain comprises SEQ ID NO: 241; or (n) the alpha or gamma chain comprises SEQ ID NO: 251 and the beta or delta chain comprises SEQ ID NO: 259; or (o) the alpha or gamma chain comprises SEQ ID NO: 269 and the beta or delta chain comprises SEQ ID NO: 277; or (p) the alpha or gamma chain comprises SEQ ID NO: 287, and the beta or delta chain comprises SEQ ID NO: 295.

[0014] In some embodiments, the TCR or antigen-binding fragment thereof recognizes a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule. In some embodiments, the MHC molecule is a human leukocyte antigen (HLA)-A molecule. In some embodiments, the HLA-A molecule is of the serotype HLA-A * 02:01. In some embodiments, the peptide epitope of HA-2 is set forth in SEQ ID NO:1.

[0015] Also provided are polynucleotides encoding any of the TCRs provided herein or antigen-binding fragments thereof, or the Vα, Vγ, Vβ, Vδ, alpha chain, beta chain, gamma chain, or delta chain thereof.

[0016] In some embodiments, the polynucleotide comprises a nucleotide sequence encoding a Vα region and a nucleotide sequence encoding a Vβ region, or a nucleotide sequence encoding a Vγ region and a nucleotide sequence encoding a Vδ region; (a) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 15, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 23, or a sequence having at least 90% sequence identity thereto; or (b) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 33, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 41, or a sequence having at least 90% sequence identity thereto; or (c) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 51, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 59, or a sequence having at least 90% sequence identity thereto; or (d) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 69, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 77, or a sequence having at least 90% sequence identity thereto; or (e) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 87, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 95, or a sequence having at least 90% sequence identity thereto; or (f) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 105, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 113, or a sequence having at least 90% sequence identity thereto; or (g) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 123, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 131, or a sequence having at least 90% sequence identity thereto; or (h) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 141, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 149, or a sequence having at least 90% sequence identity thereto; or (i) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 159, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 167, or a sequence having at least 90% sequence identity thereto; or (j) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 177, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 185, or a sequence having at least 90% sequence identity thereto; or (k) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 195, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 203, or a sequence having at least 90% sequence identity thereto; or (l) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 213, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 221, or a sequence having at least 90% sequence identity thereto; or (m) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 231, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 239, or a sequence having at least 90% sequence identity thereto; or (n) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 249, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 257, or a sequence having at least 90% sequence identity thereto; or (o) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 267, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 275, or a sequence having at least 90% sequence identity thereto; or (p) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 285, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 293, or a sequence having at least 90% sequence identity thereto.

[0017] In some embodiments, one or more of the nucleic acid sequences encoding the Vα, Vγ, Vβ, Vδ, α constant, γ constant, β constant, or δ constant chains are codon optimized. (a) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 16 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 24; or (b) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 34 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 42; or (c) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 52 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 60; or (d) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 70 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 78; or (e) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 88 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 96; or (f) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 106 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 114; or (g) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 124 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 132; or (h) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 142 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 150; or (i) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 160 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 168; or (j) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 178 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 186; or (k) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 196 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 204; or (l) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 214 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 222; or (m) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 232 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 240; or (n) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 250 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 258; or (o) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 268 and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 276; or (p) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 286, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 294.

[0018] In some embodiments, the polynucleotide comprises a nucleotide sequence encoding an alpha chain and a nucleotide sequence encoding a beta chain, or a nucleotide sequence encoding a gamma chain and a nucleotide sequence encoding a delta chain; (a) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 18, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 26, or a sequence having at least 90% sequence identity thereto; or (b) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 36, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 44, or a sequence having at least 90% sequence identity thereto; or (c) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:54, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:62, or a sequence having at least 90% sequence identity thereto; or (d) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 72, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 80, or a sequence having at least 90% sequence identity thereto; or (e) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:90, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:98, or a sequence having at least 90% sequence identity thereto; or (f) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 108, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 116, or a sequence having at least 90% sequence identity thereto; or (g) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 126, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 134, or a sequence having at least 90% sequence identity thereto; or (h) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 144, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 152, or a sequence having at least 90% sequence identity thereto; or (i) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 162, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 170, or a sequence having at least 90% sequence identity thereto; or (j) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 180, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 188, or a sequence having at least 90% sequence identity thereto; or (k) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 198, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 206, or a sequence having at least 90% sequence identity thereto; or (l) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 216, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 224, or a sequence having at least 90% sequence identity thereto; or (m) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 234, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 242, or a sequence having at least 90% sequence identity thereto; or (n) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 252, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 260, or a sequence having at least 90% sequence identity thereto; or (o) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 270, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 278, or a sequence having at least 90% sequence identity thereto; or (p) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 288, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 296, or a sequence having at least 90% sequence identity thereto. Any of the above sequences can be codon-optimized. Codon optimization can be in the variable region, the constant region, or both. Exemplary variable region-optimized codon sequences are provided in Table 3.

[0019] In some embodiments, (a) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 18 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 26; or (b) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 36 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 44; or (c) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:54 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:62; or (d) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 72 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 80; or (e) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 90 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 98; or (f) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 108 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 116; or (g) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 126 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 134; or (h) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 144 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 152; or (i) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 162 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 170; or (j) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 180 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 188; or (k) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 198 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 206; or (l) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 216 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 224; or (m) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 234 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 242; or (n) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 252 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 260; or (o) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 270 and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 278; or (p) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:288, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:296. Any of the above sequences can be codon-optimized. Codon optimization can be in the variable region, the constant region, or both. Exemplary variable region-optimized codon sequences are provided in Table 3.

[0020] In some embodiments, the nucleotide sequence encoding the alpha chain and the nucleotide sequence encoding the beta chain are present on a single nucleic acid or vector. In some embodiments, the nucleotide sequence encoding the alpha chain and the nucleotide sequence encoding the beta chain are present on a single nucleic acid or vector and expressed from a single promoter. In some embodiments, the nucleotide sequence encoding the alpha chain and the nucleotide sequence encoding the beta chain are separated by a peptide sequence that causes ribosome skipping and are present on a single nucleic acid or vector that is expressed from a single promoter. The peptide that causes ribosome skipping can be, but is not limited to, a 2A peptide. The 2A peptide can be, but is not limited to, a P2A peptide. In some embodiments, the P2A peptide comprises SEQ ID NO: 309.

[0021] In some embodiments, the nucleotide sequence encodes the amino acid sequence of SEQ ID NO: 27, 314, 45, 63, 81, 316, 99, 117, 135, 153, 318, 171, 189, 207, 225, 243, 261, 279, or 297. In some embodiments, the nucleotide sequences encoding the alpha and beta chains of SEQ ID NO: 27, 45, 63, 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, or 297 are swapped.

[0022] In some embodiments, the nucleotide sequence comprises SEQ ID NO: 28, 46, 64, 82, 100, 118, 136, 154, 172, 190, 208, 226, 244, 262, 280, 298, 315, 317, or 319, or a codon-optimized variant of SEQ ID NO: 28, 46, 64, 82, 100, 118, 136, 154, 172, 190, 208, 226, 244, 262, 280, 298, 315, 317, or 319.

[0023] Also provided herein is a vector comprising any of the polynucleotides provided herein. In some embodiments, the vector is a viral vector. In some embodiments, the viral vector is a lentiviral vector.

[0024] Also provided herein are engineered cells comprising any of the TCRs or antigen-binding fragments thereof provided herein. Also provided herein are engineered cells comprising any of the polynucleotides provided herein or any of the vectors provided herein. In some embodiments, the TCR or antigen-binding fragment thereof is heterologous to the cell. In some embodiments, the engineered cell is a cell line. In some embodiments, the engineered cell is a primary cell obtained from a subject. In some embodiments, the engineered cell is a T cell.

[0025] Also provided herein are methods for producing engineered cells that involve introducing any of the polynucleotides provided herein, or any of the vectors provided herein, into a cell to form an engineered cell.

[0026] Also provided herein are compositions comprising any of the TCRs or antigen-binding fragments thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, or any of the engineered cells provided herein. In some embodiments, the composition also comprises a pharmaceutically acceptable excipient.

[0027] Also provided herein are methods for identifying a TCR that targets a relatively hematopoietic-restricted minor histocompatibility antigen (miHA), the method involving identifying a functional TCR that recognizes a relatively hematopoietic-restricted miHA among a plurality of functional TCRs, the plurality of functional TCRs being encoded by a plurality of functional TCR-encoding nucleic acid vectors generated by a high-throughput nucleic acid amplification and assembly method using nucleic acid obtained from a single T cell among the plurality of T cells, the plurality of T cells being derived from a donor cancer patient. In some embodiments, the donor is a donor of a cancer patient that recognizes a HA-2(V) miHA. + and HLA-A * 02:01 + In some embodiments, the donor cancer patient is a recovered HA-2(V) + In some embodiments, the donor cancer patient is a (HA-2(V) - Recovered HA-2(V) undergoing AlloSCT + I am a cancer patient.

[0028] Also provided herein are methods for identifying a TCR that targets a relatively hematopoietic-restricted miHA, the method comprising: (i) generating nucleic acid vectors encoding a plurality of functional TCRs by a high-throughput nucleic acid amplification and assembly method using nucleic acid obtained from a single T cell among a plurality of T cells, the T cell being from a human donor cancer patient; and (ii) identifying a functional TCR that recognizes a relatively hematopoietic-restricted miHA from among the plurality of functional TCRs encoded by the nucleic acid vector encoding the plurality of functional TCRs. In some embodiments, the donor has HA-2(V) + and HLA-A * 02:01 + is.

[0029] In some embodiments, the relatively hematopoietic-restricted miHA is the minor histocompatibility antigen HA-2. In some embodiments, the identified functional TCR recognizes a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule. In some embodiments, the MHC molecule is a human leukocyte antigen (HLA)-A molecule. In some embodiments, the HLA-A molecule is of the serotype HLA-A * 02:01. In some embodiments, the peptide epitope of HA-2 is set forth in SEQ ID NO:1.

[0030] In some embodiments, T cells from a human cancer patient are cultured under conditions for cell expansion of the T cells prior to generation of multiple functional TCR-encoding nucleic acid vectors.

[0031] In some embodiments, the T cells from a human cancer patient are not cultured under conditions for cell expansion of the T cells prior to generation of a plurality of functional TCR-encoding nucleic acid vectors.

[0032] In some embodiments, high-throughput nucleic acid amplification and assembly involves (1) amplifying a first amplification product and a second amplification product from complementary DNA (cDNA) generated from RNA obtained from a single T cell among a plurality of T cells sorted into each of a plurality of distinct locations on a device, wherein the first amplification product comprises a nucleotide sequence encoding a full-length Vα region or a full-length Vγ region of a TCR, and the second amplification product comprises a nucleotide sequence encoding a full-length Vβ region or a full-length Vδ region of a TCR; and (2) assembling the first amplification product and the second amplification product from each of the plurality of distinct locations into a nucleic acid vector to obtain an assembled nucleic acid vector comprising a nucleotide sequence encoding a functional TCR for each of the plurality of distinct locations, wherein the functional TCR comprises (i) a full-length Vα region and a full-length Vβ region derived from the single T cell, or (ii) a full-length Vγ region and a full-length Vδ region derived from the single T cell.

[0033] Also provided herein are engineered cells comprising a TCR identified by any of the methods described herein. In some embodiments, the engineered cells are T cells modified to knock out endogenously expressed TCRs. Also provided herein are compositions comprising any of the engineered cells provided herein. In some embodiments, the composition also comprises a pharmaceutically acceptable excipient.

[0034] Also provided herein are therapeutic methods involving administering any of the TCRs or antigen-binding fragments thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein to a subject with a disease or disorder. Also provided herein are any of the TCRs or antigen-binding fragments thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein for use in treating a disease or disorder in a subject. Also provided herein is the use of any of the TCRs or antigen-binding fragments thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein in the manufacture of a medicament for the treatment of a disease or disorder in a subject. Also provided herein is the use of any of the TCRs or antigen-binding fragments thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein for the treatment of a disease or disorder in a subject.

[0035] In some embodiments, the subject is eligible for or will undergo allogeneic hematopoietic stem cell transplantation (HSCT). * The subject is eligible for or will undergo allogeneic hematopoietic stem cell transplantation (HSCT) from a donor that does not express 02:01. In some embodiments, the subject has or has been diagnosed with a malignant hematologic disorder. In some embodiments, the subject has or has been diagnosed with acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), or acute lymphoblastic leukemia (ALL). In some embodiments, the subject has or has been diagnosed with a liquid tumor, hematopoietic tumor, lymphoma, or chronic myeloid leukemia (CML). In some embodiments, administration of the engineered cells or compositions induces or enhances cell death of cells associated with a malignant hematological disorder or induces or enhances a graft versus leukemia effect (GVL) in a subject. [Brief explanation of the drawings]

[0036] [Figure 1A] Graph showing killing of LCL 00177 (HA-2 M / M) and LCL 00174 (HA-2 V / V) cells by engineered T cells expressing TCR D, TCRA, or TCR H. The left bar in each pair represents LCL 00177 cells. The right bar in each pair represents LCL 00174 cells. [Figure 1B]Graph showing killing of LCL 00177 (HA-2 M / M) and LCL 00174 (HA-2 V / V) cells by engineered T cells expressing an exemplary anti-HA-2 TCR or control T cells. The left bar in each pair represents LCL 00177 non-immune cells. The right bar in each pair represents LCL 00174 immune cells. DETAILED DESCRIPTION OF THE INVENTION

[0037] Provided herein are TCRs, including recombinant TCRs, that bind to or recognize peptide epitopes associated with relatively hematopoietically restricted minor histocompatibility antigens, e.g., HA-2, such as peptide epitopes expressed on the surface of cells in the context of MHC molecules. Among the provided embodiments are approaches useful in the treatment of such diseases and conditions and / or for targeting cell types, such as cancer cells or cells associated with hematological ailments. In some embodiments, the provided TCRs and antigen-binding fragments thereof bind to or recognize peptide epitopes of HA-2 in the context of MHC molecules.

[0038] Also provided herein are nucleic acid molecules encoding TCRs, engineered cells containing TCRs, compositions containing TCRs or cells, and methods of treatment or use, such as therapeutic applications, involving administering such TCRs, engineered cells, or compositions, as well as uses of such TCRs, cells, or compositions. In some embodiments, engineered cells expressing the provided TCRs or antigen-binding fragments thereof exhibit cytotoxic activity against target cells expressing the peptide epitope, such as cancer cells or cells associated with hematologic diseases. Also provided herein are methods for identifying TCRs that target relatively hematopoietic-restricted miHAs.

[0039] Allogeneic stem cell transplantation (Allo-SCT) can be a curative treatment for patients with hematologic malignancies as well as for patients with nonmalignant but medically serious conditions, such as hemoglobinopathies, thalassemias, and autoimmune diseases. Allo-SCT can also be used to create tolerance to transplanted solid organs. Mature αβ T cells contained in the donor allograft play a key role and can be considered in two broad classes. One class promotes the reconstitution of anti-pathogen immunity, particularly through the transfer of memory T cells. The second class of T cells, called alloreactive T cells, recognizes the patient as "non-self." When allo-SCT is used to treat hematologic malignancies, alloreactive donor T cells can kill malignant cells, thereby mediating the graft-versus-host leukemia (GVL) effect. However, they can also cause graft-versus-host disease (GVHD), in which alloreactive T cells attack healthy host tissues, including, for example, the skin, gut, and liver.

[0040] In human leukocyte antigen (HLA)-matched or haploidentical alloSCT, alloreactive T cells target miHA, a peptide product of a coding polymorphism that distinguishes the recipient from the donor. Importantly, alloreactive CD8+ T cells targeting miHA, which has expression restricted to hematopoietic cells, are less likely to cause GVHD. Administration of anti-miHA T cells, among other strategies, could minimize the risk of extensive toxicity without compromising therapeutic efficacy, either in the context of boosting alloSCT or as a stand-alone therapy.

[0041] Cell therapies (including those involving the administration of cells expressing recombinant receptors or TCRs specific for a disease or disorder of interest, such as recombinant TCRs and / or other recombinant antigen receptors), as well as other adoptive immune cell and adoptive T cell therapies, can be effective in treating diseases and disorders. In certain circumstances, available approaches to adoptive cell therapy may not always be entirely satisfactory. In some situations, optimal effectiveness may depend on the ability of the administered cells to express the recombinant receptor and the ability of the recombinant receptor to recognize and bind to a target antigen within the subject, such as a peptide epitope of HA-2, based, for example, on the affinity of the antigen-binding domain of the TCR for that target antigen. In some cases, the consistency and / or efficiency of expression of the recombinant receptor, as well as the activity of the receptor, limit the availability of certain cells or certain cell populations to certain therapeutic approaches.

[0042] When engineered into a human T cell, a xenogeneic TCR (e.g., a humanized TCR or fully human recombinant TCR) can compete with the endogenous TCR complex and / or form mispairings with endogenous TCR chains, which, in certain aspects, can reduce recombinant TCR signaling, activity, and / or expression, ultimately resulting in reduced activity of the engineered cell. For example, in some cases, suboptimal expression of an engineered or recombinant TCR can result from competition with endogenous TCRs and / or TCRs with mispairing chains for signaling molecules and / or domains involved in enabling expression of the complex on the cell surface, such as the invariant CD3 signaling molecule (e.g., availability of co-expressed CD3 delta, epsilon, gamma, and / or zeta chains). In some aspects, available CD3 molecules can limit TCR expression and function within the cell. To reduce or eliminate mispairing between a xenogeneic TCR and an endogenous TCR, expression of the endogenous TCR can be reduced or eliminated. In some embodiments, the anti-HA-2 TCR-expressing T cells are further modified to knock out endogenously expressed TCRs, hi some embodiments, expression of the endogenous TRAC gene and the endogenous TRBC gene in the T cells is knocked out, such as by CRISPR.

[0043] In some aspects, provided embodiments are based on the observation of improved affinity, expression, or activity of exemplary fully human recombinant TCRs, such as certain provided TCRs specific for HA-2, even at low effector-to-target (E:T) ratios. The activity, e.g., cytokine secretion and / or cytolytic activity, of engineered T cells expressing the recombinant TCRs can, in some cases, be limited when there are fewer engineered T cells compared to target cells. In some aspects, such improved activity, particularly at low E:T ratios and using fully human sequences, is advantageous for improving therapeutic efficacy.

[0044] In some cases, certain available approaches for obtaining antigen-specific recombinant receptors, such as recombinant TCRs, may result in recombinant receptors that exhibit cross-reactivity to different non-target antigens (see, e.g., Cameron et al., (2013) Science Translational Medicine, 5(197):197ral03). In some aspects, provided embodiments may be used in combination with other antigen-specific recombinant receptors, such as those targeting HLA subtypes A, B, C, D, E, E, F, F, HLA subtypes B, E, F, F, HLA subtypes C, E, F, F, HLA subtypes D, E, F, F, HLA subtypes ... * This is based on the observation that certain provided TCRs specific for particular immunogenic HA-2 peptides presented by 02:01 do not exhibit cross-reactivity to cells expressing other peptide antigens or alloreactivity to other HLA subtypes. Thus, the provided TCRs exhibit improved expression and activity while minimizing the risk of cross-reactivity to other antigens, such as non-target antigens or peptide epitopes presented via non-target HLA subtypes, that may be present in the subject.

[0045] In some aspects, therapeutic approaches using such TCRs, e.g., adoptive cell therapy with engineered human T cells expressing the provided recombinant TCRs, may ultimately result in increased efficacy, e.g., by improving the GVL effect. In some situations, provided embodiments, including TCRs, polynucleotides encoding such TCRs, engineered cells, and cell compositions, may offer various advantages over available TCR-based therapies, improving the activity of recombinant receptors and responses to adoptive cell therapy targeting cancer cells and cells associated with hematological diseases.

[0046] All publications, including patent documents, scientific articles, and databases, referenced in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. To the extent that a definition set forth herein is contrary to or otherwise inconsistent with a definition set forth in a patent, application, published application, or other publication incorporated herein by reference, the definition set forth herein takes precedence over the definition incorporated herein by reference.

[0047] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0048] I. T cell receptors targeting histocompatibility antigen 2 (HA-2) Provided herein are TCRs, such as those that bind to or recognize peptide epitopes associated with relatively hematopoietic-restricted miHAs in the context of MHC molecules, e.g., peptide epitopes expressed on the surface of immune cells, cancer cells, and / or cells associated with hematologic diseases. In some embodiments, the provided TCRs bind to or recognize peptide epitopes of miHA HA-2 in the context of MHC molecules. Such TCRs and antigen-binding fragments exhibit antigen specificity that binds to or recognizes such peptide epitopes. Also provided in some embodiments are nucleic acid molecules encoding TCRs, engineered cells expressing TCRs, compositions, and therapeutic methods involving administering such TCRs, engineered cells, or compositions. In some aspects, engineered cells expressing the provided TCRs or antigen-binding fragments exhibit cytotoxic activity against target cells expressing the peptide epitopes, such as cancer cells or cells associated with hematologic diseases.

[0049] In some embodiments, the TCRs described recognize the miHA HA-2 antigen expressed on bone marrow-derived cells from humans with the HA-2 YIGEVLVSV (SEQ ID NO: 1) allele. The HA-2 antigen is encoded by the myosin 1G (MYO1G) gene.

[0050] A. Allogeneic stem cell transplantation AlloSCT is a potentially curative treatment option for patients with hematological malignancies. In alloSCT (HLA-matched or haploidentical), patients undergo a pre-transplant regimen consisting of chemotherapy, sometimes with radiation therapy, which facilitates engraftment by killing any remaining malignant cells, creating space in the recipient's bone marrow for donor stem cell engraftment, and by killing the patient's immune cells that can mediate donor allograft rejection.

[0051] Mature alpha / beta donor T cells contained in the donor graft play important roles and can be considered in two broad classes. One class promotes the reconstitution of anti-pathogen immunity, particularly through the transfer of memory T cells. The second class of T cells recognizes the patient as "non-self." These so-called "allo-reactive" T cells have both positive and detrimental effects. An important advantage of these cells is their ability to kill recipient malignant cells, which mediate the GVL effect. Alloreactive T cells can also kill normal patient or host hematopoietic and immune cells, both of which create space for cell engraftment and reduce immunological rejection of donor cells. However, donor T cells can also attack normal non-hematopoietic recipient tissues, causing GVHD. Therefore, patients receiving alloSCT alone undergo some form of systemic immunosuppression to reduce the frequency and severity of GVHD.

[0052] Despite the GVL effect, recurrent malignancies are the single largest cause of treatment failure and death in recipients of alloSCT for the treatment of hematologic neoplasms. There is good reason to believe that engineering more effective alloreactive T cell responses could reduce relapse. GVHD and the consequences of systemic immunosuppression (e.g., infection) are other major causes of morbidity and mortality. These, too, could be mitigated if alloreactivity were better engineered to focus on hematopoietic cells rather than normal host tissues. Despite these limitations, alloSCT is the global standard of care for patients with intermediate- to high-risk hematologic malignancies, supported by data from multiple sources supporting higher survival rates than without transplantation.

[0053] In HLA-matched or haploidentical alloSCT, alloreactive donor T cells target miHAs expressed in the recipient. MiHAs are peptide products of coding polymorphisms that distinguish the recipient from the donor. These polymorphisms exist at stable, known frequencies in the population and are inherited by Mendelian genetics. Some genes encoding miHAs are expressed in a wide range of tissues, while others are relatively restricted to hematopoietic cells. As currently practiced, there is no control over which miHAs are targeted in alloSCT. Because T cell responses against miHAs expressed on normal tissues almost always occur, severe GVHD is a major risk, thus necessitating immunosuppression. In contrast, miHAs, which are expressed relatively restricted to hematopoietic cells, are considered ideal targets for immunotherapy with donor-derived CD8+ T cells in combination with alloSCT. CD8+ T cells targeting such antigens can kill recipient malignant blood cells, mediating the GVL effect. T cells targeting relatively hematopoietic-restricted antigens can mediate graft-versus-host leukemia and promote engraftment with a reduced risk of graft-versus-host disease. They can also kill non-malignant recipient hematopoietic cells, including immune cells, thereby promoting engraftment and reducing the risk of immune rejection. Interestingly, CD8+ T cells targeting relatively hematopoietic-restricted miHAs have a reduced risk of causing GVHD.

[0054] CD8+ T cells recognize their targets through their antigen receptors (TCRs). The process of generating these receptors creates a highly diverse repertoire of unique TCRs, with each person having approximately 10 7 It is estimated that TCRs contain T cells that express more than 100 unique receptors. This diversity allows individuals to respond to a wide range of pathogens as well as miHAs. Unlike antibodies, which bind intact proteins, TCRs recognize short peptides, usually about 8-12 amino acids long, embedded on the surface of MHC molecules expressed on the surface of cells.

[0055] The advantage of this system of antigen detection is that T cells can recognize peptides derived from any protein, even those not expressed on the cell surface. Throughout evolution, MHC molecules have diversified in populations, with most of the variation occurring in the portion of the MHC molecule that binds and displays or presents peptides to the TCR. This allows for a large diversity of peptides that can be presented to T cells by MHC molecules, with the preference or restriction of certain peptides to specific MHC molecules. The result is that T cell presentation and recognition of each miHA is generally restricted to a single MHC type. Importantly, over the past several decades, over 50 relatively hematopoietic-restricted miHAs have been identified, a number sufficient to ensure that nearly all donor / recipient combinations, regardless of MHC type, have targetable relatively hematopoietic-restricted miHAs.

[0056] B. Histocompatibility antigen HA-2 HA-2 is a hematopoietically expressed, relatively hematopoietic-restricted miHA that has also been found on Hofbauer cells and trophoblast cells in the human placenta. HA-2 can be recognized and responded to under certain genetic conditions in the context of bone marrow transplantation. The antigenic peptide derived from HA-2 results from a single nucleotide difference between a non-immunogenic peptide (the "M peptide") containing the amino acid sequence YIGEVLVSM (SEQ ID NO: 2) and an immunogenic peptide (the "V peptide") containing the amino acid sequence YIGEVLVSV (SEQ ID NO: 1). The immunogenic peptide binds to class I MHC molecules, HLA-A, and HLA-B. * 02:01 context. HLA-A on antigen-presenting cells (APCs) as a result of immunogenic single nucleotide polymorphisms (SNPs). * 02:01 peptide-binding groove, thus increasing the binding affinity of HA-2 V peptides to HLA-A * In some embodiments, the subject is provided with an immunogenic peptide that can be recognized by 02:01-restricted T cells. *have received, are eligible for, or will undergo allogeneic hematopoietic stem cell transplantation (HSCT) from a donor that does not express 02:01, as such donor cells will not be recognized by any T cells with a TCR that recognizes the V peptide version of HA-2 and therefore will not be eliminated by such engineered cells.

[0057] The difference in immunogenicity of these peptides may be due to various factors. For example, the proteasomal cleavage and TAP-mediated transport of the two peptides into the endoplasmic reticulum are similar, and both variants can bind to HLA-restricted alleles. However, the M peptide does not bind to HLA-A. * 02:01 HLA-A, possibly related to the relatively large size of the arginine residue, which poses steric and electrostatic hindrance to the D-pocket residues. * It has lower affinity and less stable binding to 02:01. Differences in both MHC molecule and TCR binding may explain the immunogenicity of the HA-2 V peptide.

[0058] In some instances, the miHA HA-2 V peptide (YIGEVLVSV; SEQ ID NO: 1) is targeted, while the non-immunogenic M peptide (YIGEVLVSM; SEQ ID NO: 2) is not. HA-2 is a preferred target because its expression is relatively restricted to hematopoietic cells, is presented (or "restricted") by the most common human HLA types, and has an allele frequency of approximately 50%. See de Bueger et al. (1992) J Immunol Baltim Md 1950 149(5):1788-1794 and Wilke et al. (2003) Hematol J. 4(5):315-320.

[0059] In some embodiments, the TCR recognizes or binds to the HA-2 epitope in the context of an MHC molecule, such as an MHC class I molecule. In some embodiments, the MHC class I molecule is any one or more subtypes thereof, e.g., HLA-A *In some cases, there may be differences in the frequency of subtypes between different populations. For example, in some embodiments, more than 95% of the HLA-A2 positive Caucasian population is HLA-A2. * 02:01, whereas in the Chinese population, the frequency is approximately 23% HLA-A * In some embodiments, the MHC molecule is HLA-A * It is 02:01.

[0060] In some aspects, the provided TCRs or antigen-binding fragments thereof recognize or bind to a non-immunogenic epitope or domain of HA-2, such as a non-immunogenic V peptide comprising the amino acid sequence YIGEVLVSV (SEQ ID NO: 1). In some embodiments, the TCRs recognize or bind to an HLA-A * 02:01 + derived from a TCR donor subject that is HLA-A * 02:01 + For use in combination with alloSCT of the recipient subject, the alloSCT graft is HLA-A * 02:01 - It originates from a transplant donor who is

[0061] In some embodiments, the TCR or antigen-binding fragment thereof is isolated or purified, or recombinant. In certain embodiments, any of the provided TCRs or antigen-binding fragments thereof is recombinant. In some aspects, the TCR or antigen-binding fragment thereof is human. In some aspects, the TCR is a single chain. In other embodiments, the TCR contains two chains. In some embodiments, the TCR or antigen-binding fragment thereof is expressed on the surface of a cell (e.g., a T cell, such as a T cell engineered to lack expression of an endogenous TCR).

[0062] In some embodiments, the provided TCRs have one or more specified functional characteristics, such as a binding characteristic including binding to a particular epitope, and / or a particular binding affinity, e.g., as described herein. In some embodiments, engineered cells, such as T cells, expressing a provided TCR have one or more specified functional characteristics, such as a binding characteristic including binding to a particular epitope, a particular binding affinity, activation or stimulation of cell signaling, such as T cell signaling or TCR signaling, secretion of cytokines, and / or killing of target cells expressing or presenting an antigen, e.g., as described herein.

[0063] In some embodiments, the provided binding molecule is a TCR or an antigen-binding fragment thereof. In some embodiments, the TCR is an alpha chain comprising a Vα region and a beta chain comprising a Vβ region (also known as TCRα and TCRβ, respectively), or a gamma chain comprising a Vγ region and a delta chain comprising a Vδ region (also known as TCRγ and TCRδ, respectively), or a molecule containing an antigen-binding portion thereof, which can specifically bind to an antigen, e.g., a peptide antigen or peptide epitope bound to an MHC molecule. In some embodiments, the TCR is in the αβ form (e.g., an αβ TCR). In some embodiments, the TCR is in the γδ form (e.g., a γδ TCR). Typically, TCRs present in the αβ or γδ form are generally structurally similar, although T cells expressing them may have different anatomical locations or functions. TCRs can be found on the surface of cells or in a soluble form. Generally, TCRs are found on the surface of T cells, where they are responsible for recognizing antigens, typically peptides bound to MHC molecules.

[0064] In some embodiments, the TCRs provided herein can be intact or full-length TCRs, such as TCRs containing a full-length α chain and a full-length β chain, or a TCR containing a full-length γ chain and a full-length δ chain. In some embodiments, the antigen-binding portion of a TCR provided herein can be less than a full-length TCR, but binds to a specific peptide bound to an MHC molecule, such as binding to an MHC-peptide complex. In some cases, the antigen-binding portion or fragment of a TCR can contain only a portion of the structural domain of a full-length or intact TCR, but can still bind to a peptide epitope, such as an MHC-peptide complex, to which a full-length TCR binds. In some cases, the antigen-binding portion contains a variable domain of a TCR, such as the Vα and Vβ regions of a TCR provided herein or the Vγ and Vδ regions of a TCR, but is sufficient to form a binding site for the antigen-binding portion to bind to a specific MHC-peptide complex.

[0065] In some embodiments, the variable domain of a TCR generally contains the CDRs, which contribute to the antigen recognition and binding capacity and specificity of peptides, MHC molecules, and / or MHC-peptide complexes. In some embodiments, the CDRs of a TCR, or a combination thereof, form all or substantially all of the antigen-binding site of a given TCR molecule. The various CDRs within the variable region of a TCR chain are generally separated by framework regions (FRs), which generally show less variability among TCRs compared to the CDRs (see, e.g., Jores et al., Proc. Nat'l Acad. Sci. USA 87:9138, 1990; Chothia et al., EMBO J. 7:3745, 1988; see also Lefranc et al., Dev. Comp. Immunol. 27:55 (2003)). In some embodiments, CDR-3 is the primary CDR responsible for antigen binding or specificity, or the most important of the three CDRs on a given TCR variable region for antigen recognition and / or interaction with the processed peptide portion of a peptide-MHC complex. In some circumstances, CDR-1 of the alpha chain can interact with the N-terminal portion of a particular antigenic peptide. In some circumstances, CDR-1 of the beta chain can interact with the C-terminal portion of a peptide. In some circumstances, CDR-2 is the most strongly contributing or primary CDR responsible for interaction with or recognition of the MHC portion of an MHC-peptide complex. In some embodiments, the variable region of the beta chain can contain an additional hypervariable region (e.g., CDR4 or HVR4), which is generally involved in superantigen binding and not antigen recognition (Kotb (1995) Clinical Microbiology Reviews, 8:411-426).

[0066] In some embodiments, the α and / or β chains of the TCR, or the γ and / or δ chains of the TCR, may also contain constant domains, transmembrane domains, and / or short cytoplasmic tails (see, e.g., Janeway et al., Immunobiology: The Immune System in Health and Disease, 3 rd Ed., Current Biology Publications, p. 4:33, 1997). In some aspects, each chain (e.g., alpha or beta) of the TCR can have one N-terminal immunoglobulin variable domain, one immunoglobulin constant domain, a transmembrane region, and a short C-terminal cytoplasmic tail. In some embodiments, the TCR associates, e.g., via the cytoplasmic tail, with the invariant protein of the CD3 complex, which is involved in mediating signal transduction. In some cases, the structure allows the TCR to associate with other molecules, such as CD3 and its subunits. For example, a constant domain-containing TCR with a transmembrane region can anchor the protein to the cell membrane and associate with the invariant subunit of the CD3 signaling apparatus or complex. The intracellular tails of the CD3 signaling subunits (e.g., CD3γ, CD3δ, CD3ε, and CD3ζ chains) contain one or more immunoreceptor tyrosine-based activation motifs or ITAMs and are generally involved in the signaling capabilities of the TCR complex.

[0067] Various domains or regions of the TCR can be identified. In some cases, the exact location of a domain or region may vary depending on the particular structural or homology modeling or other features used to describe a particular domain. It is understood that reference to amino acids, including particular sequences set forth as SEQ ID NOs, used to describe the domain organization of a TCR is for illustrative purposes and is not meant to limit the scope of the provided embodiments. In some cases, a particular domain (e.g., a variable domain or a constant domain) may be longer or shorter than a few amino acids (e.g., one, two, three, or four). In some aspects, residues of a TCR are known or can be identified according to the International Immunogenetics Information System (IMGT) numbering system (see, e.g., www.imgt.org; see also Lefranc et al. (2003) Developmental and Comparative Immunology, 27(1);55-77; and The T Cell Factsbook 2nd Edition, Lefranc and LeFranc Academic Press 2001). Using this system, the CDR-1 sequence within the TCR Vα and / or Vβ regions optionally corresponds to amino acids located between residues 27 and 38 (inclusive), the CDR-2 sequence within the TCR Vα and / or Vβ regions optionally corresponds to amino acids located between residues 56 and 65 (inclusive), and the CDR-3 sequence within the TCR Vα and / or Vβ regions optionally corresponds to amino acids located between residues 105 and 117 (inclusive).

[0068] In some embodiments, some TCRs or antigen-binding fragments thereof provided herein bind to or recognize, in the context of an MHC molecule, a relatively hematopoietically restricted minor histocompatibility antigen, such as HA-2. In some embodiments, some TCRs or antigen-binding fragments thereof provided herein recognize or bind to a non-immunogenic epitope or domain of HA-2, such as, for example, an immunogenic V peptide comprising the amino acid sequence YIGEVLVSV (SEQ ID NO: 1). In some embodiments, some TCRs or antigen-binding fragments thereof provided herein do not recognize or bind to a non-immunogenic epitope or domain of HA-2, such as, for example, a non-immunogenic M peptide comprising the amino acid sequence YIGEVLVSM (SEQ ID NO: 2). In some embodiments, some TCRs or antigen-binding fragments thereof provided herein preferentially or selectively recognize or bind to immunogenic epitopes or domains of HA-2, such as the immunogenic V peptide comprising the amino acid sequence YIGEVLVSV (SEQ ID NO: 1), and some TCRs or antigen-binding fragments thereof do not recognize or bind to non-immunogenic epitopes or domains of HA-2, such as the non-immunogenic M peptide comprising the amino acid sequence YIGEVLVSM (SEQ ID NO: 2), or exhibit reduced affinity for binding to non-immunogenic epitopes and therefore increased relative selectivity for binding to immunogenic epitopes.

[0069] In some aspects, some of the TCRs or antigen-binding fragments thereof provided herein are HLA-A * These include those that bind to or recognize an epitope of HA-2 complexed with an MHC molecule of a specific HLA type, such as 02:01, such as an immunogenic V peptide comprising the amino acid sequence YIGEVLVSV (SEQ ID NO: 1).

[0070] In some embodiments, the TCRs provided herein are full-length TCRs. In some embodiments, the TCRs provided herein are dimeric TCRs (dTCRs). In some embodiments, the TCRs provided herein are single-chain TCRs (scTCRs). The TCRs provided herein can be in cell-associated or soluble form. In some embodiments, the TCRs provided herein are cell-associated forms that are expressed on the surface of a cell (e.g., a T cell, such as a T cell engineered to lack expression of an endogenous TCR).

[0071] In some embodiments, the TCRs provided herein are scTCRs, which are single amino acid chains containing an α chain and a β chain capable of binding to an MHC-peptide complex. Typically, scTCRs can be generated as described elsewhere. See, e.g., WO 96 / 13593, WO 96 / 18105, WO 99 / 18129, WO 04 / 033685, WO 2006 / 037960, WO 2011 / 044186; U.S. Patent No. 7,569,664; and Schlueter, CJ et al. J. Mol. Biol. 256, 859 (1996).

[0072] C. Exemplary Variable Domains Provided herein are TCRs or antigen-binding fragments thereof that recognize or bind to an epitope or region of a relatively hematopoietically restricted minor histocompatibility antigen, such as HA-2, in the context of an MHC molecule. In some embodiments, the HA-2 peptide is the YIGEVLVSV (SEQ ID NO: 1) peptide. The HA-2 antigen is encoded by the myosin 1G (MYO1G) gene. Exemplary sequences (e.g., CDRs, Vα and / or Vβ, or Vγ and / or Vδ, and constant region sequences) of HA-2-specific TCRs are provided.

[0073] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein bind to or recognize an immunogenic HA-2 allele displayed on the surface of leukemia cells in an alloSCT recipient. In some aspects, the cytotoxic activity of T cells expressing an anti-HA-2 TCR is stimulated upon contact of the T cells with a target cell that presents or expresses an antigen, such as an immunogenic HA-2 peptide. In some embodiments, some of the TCRs or antigen-binding fragments provided herein bind to or recognize a peptide epitope of HA-2 (e.g., a peptide epitope of an immunogenic allele of HA-2) in the context of an MHC, such as a particular MHC or a particular HLA subtype.

[0074] Among such TCRs or antigen-binding fragments thereof are TCRs or antigen-binding fragments thereof that contain either the Vα and Vβ regions, or the Vγ and Vδ regions, the sequences individually described, or a sufficient antigen-binding portion of such a sequence. In some embodiments, the provided TCRs or antigen-binding fragments thereof (e.g., anti-HA-2 TCRs) contain a Vα region sequence or a Vγ region sequence, or a sufficient antigen-binding portion thereof, that contains CDR-1, CDR-2, and / or CDR-3 described herein. In some embodiments, the provided TCRs or antigen-binding fragments thereof (e.g., anti-HA-2 TCRs) contain a Vβ region sequence or a Vδ region sequence, or a sufficient antigen-binding portion thereof, that contains CDR-1, CDR-2, and / or CDR-3 described herein. In some embodiments, the TCR or antigen-binding fragment thereof (e.g., an anti-HA-2 TCR) contains a Vα or Vγ region sequence containing CDR-1, CDR-2, and / or CDR-3 described herein, and contains a Vβ or Vδ region sequence containing CDR-1, CDR-2, and / or CDR-3 described herein. Also provided are TCRs having sequences that are at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences.

[0075] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein contain a Vα or Vγ region containing a CDR-3 comprising an amino acid sequence set forth in any of SEQ ID NOs: 13, 31, 49, 67, 85, 103, 121, 139, 157, 175, 193, 211, 229, 247, 265, 283, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence.

[0076] In some aspects, the TCRs or antigen-binding fragments thereof provided herein contain a Vα or Vγ region containing a CDR-3 contained within an amino acid sequence set forth in any of SEQ ID NOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence that is at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such a sequence.

[0077] In some embodiments, the Vα or Vγ region contains a CDR-1 comprising an amino acid sequence set forth in any of SEQ ID NOs: 11, 29, 47, 65, 83, 101, 119, 137, 155, 173, 191, 209, 227, 245, 263, and 281, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. In some embodiments, the Vα or Vγ region contains a CDR-1 contained within an amino acid sequence set forth in any of SEQ ID NOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. In some embodiments, the Vα or Vγ region contains a CDR-2 comprising an amino acid sequence set forth in any of SEQ ID NOs: 12, 30, 48, 66, 84, 102, 120, 138, 156, 174, 192, 210, 228, 246, 264, and 282, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. In some embodiments, the Vα or Vγ region contains a CDR-2 contained within an amino acid sequence set forth in any of SEQ ID NOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence.

[0078] In some cases, the TCRs or antigen-binding fragments thereof provided herein contain a Vβ or Vδ region containing a CDR-3 comprising an amino acid sequence set forth in any of SEQ ID NOs: 21, 39, 57, 75, 93, 111, 129, 147, 165, 183, 201, 219, 237, 255, 273, and 291, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. In some embodiments, the TCRs or antigen-binding fragments thereof provided herein contain a Vβ or Vδ region containing a CDR-3 contained within an amino acid sequence set forth in any of SEQ ID NOs: 22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, and 292, or a sequence that is at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such a sequence.

[0079] In some examples, the Vβ or Vδ region contains a CDR-1 comprising an amino acid sequence set forth in any of SEQ ID NOs: 19, 37, 55, 73, 91, 109, 127, 145, 163, 181, 199, 217, 235, 253, 271, and 289, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. In some embodiments, the Vβ or Vδ region contains a CDR-1 contained within an amino acid sequence set forth in any of SEQ ID NOs: 22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, and 292, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. In some embodiments, the Vβ or Vδ region contains a CDR-2 comprising an amino acid sequence set forth in SEQ ID NOs: 20, 38, 56, 74, 92, 110, 128, 146, 164, 182, 200, 218, 236, 254, 272, and 290, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. In some embodiments, the Vβ or Vδ region contains a CDR-2 contained within an amino acid sequence set forth in any of SEQ ID NOs: 22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 218, 236, 254, 272, and 290, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence.

[0080] In some embodiments, the Vα or Vγ region contains an amino acid sequence set forth in any of SEQ ID NOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In some examples, the Vβ or Vδ region contains an amino acid sequence set forth in any of SEQ ID NOs: 22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, and 292, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In some embodiments, the TCR contains an alpha chain comprising either such a Vα region sequence or a Vγ region sequence, and either such a Vβ region sequence or a Vδ region sequence.

[0081] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein that have the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule comprise a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 11 (or a variant of SEQ ID NO: 1 having 11 or 12 amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 12 (or a variant of SEQ ID NO: 2 having 1 or 2 amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 13 (or a variant of SEQ ID NO: 13 having 1 or 2 amino acid modifications). and a beta chain (or delta chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 19 (or a variant of SEQ ID NO: 19 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 20 (or a variant of SEQ ID NO: 20 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 21 (or a variant of SEQ ID NO: 21 with one or two amino acid modifications). Examples of such TCRs having these CDRs and having the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule include, but are not limited to, TCR A. The CDR-1, CDR-2, and CDR-3 sequences of any of TCRs B through P as identified in Table 1 may be substituted for the CDR-1, CDR-2, and CDR-3 sequences of TCR A listed above.

[0082] In some cases, an alpha chain (or gamma chain) having the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2 and having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 11 (or a variant of SEQ ID NO: 1 with 11 or 12 amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 12 (or a variant of SEQ ID NO: 2 with 1 or 2 amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 13 (or a variant of SEQ ID NO: 13 with 1 or 2 amino acid modifications). ), and a beta chain (or delta chain) having CDR-1 having the amino acid sequence set forth in SEQ ID NO: 19 (or a variant of SEQ ID NO: 19 with one or two amino acid modifications), CDR-2 having the amino acid sequence set forth in SEQ ID NO: 20 (or a variant of SEQ ID NO: 20 with one or two amino acid modifications), and CDR-3 having the amino acid sequence set forth in SEQ ID NO: 21 (or a variant of SEQ ID NO: 21 with one or two amino acid modifications), may comprise any suitable framework region.For example, such a TCR or antigen-binding fragment thereof may have a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO: 14 upstream of the amino acid sequence of SEQ ID NO: 11 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications), a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 14 between the amino acid sequences of SEQ ID NO: 11 and SEQ ID NO: 12 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications). a framework region 3 having the entire amino acid sequence set forth in SEQ ID NO: 14 located between the amino acid sequences of SEQ ID NO: 12 and SEQ ID NO: 13 (or variants of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications); and a framework region 4 having the entire amino acid sequence set forth in SEQ ID NO: 14 located downstream of the amino acid sequence of SEQ ID NO: 13 (or variants of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications). an alpha chain comprising: a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO: 22 upstream of the amino acid sequence of SEQ ID NO: 19 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications); a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 22 between the amino acid sequences of SEQ ID NO: 19 and SEQ ID NO: 20 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications); and a beta strand comprising framework region 3 having the entire amino acid sequence set forth in SEQ ID NO:22 between the amino acid sequence of SEQ ID NO:21 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), and framework region 4 having the entire amino acid sequence set forth in SEQ ID NO:22 downstream of the amino acid sequence of SEQ ID NO:21 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications).The CDR-1, CDR-2, CDR-3, and variable region sequences of any of TCRs B to P as identified in Table 1 may be substituted for the TCR A CDR-1, CDR-2, CDR-3, and variable region sequences listed above.

[0083] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise an alpha chain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 14, and a beta chain comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 22. For example, a TCR or antigen-binding fragment thereof provided herein can comprise an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 14, and a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 22. In some cases, a TCR or antigen-binding fragment thereof provided herein can comprise (a) an alpha chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 14, and (b) a beta chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 22. Any of the TCR B-P variable region sequences as identified in Table 1 can be substituted for the TCR A variable region sequences listed above.

[0084] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise: (a) an alpha chain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 14 (with the proviso that the alpha chain comprises the amino acid sequences set forth in SEQ ID NOs: 11, 12, and 13); and (b) a beta chain comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 22 (with the proviso that the beta chain comprises the amino acid sequences set forth in SEQ ID NOs: 19, 20, and 21). For example, a TCR or antigen-binding fragment thereof provided herein can comprise: (a) an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 14, with the proviso that the alpha chain comprises the amino acid sequence set forth in SEQ ID NOs: 11, 12, and 13; and (b) a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity to the amino acid sequence set forth in SEQ ID NO: 22, with the proviso that the beta chain comprises the amino acid sequence set forth in SEQ ID NOs: 19, 20, and 21. The CDR-1, CDR-2, CDR-3, and variable region sequences of any of the TCRs B through P as identified in Table 1 can be substituted for the TCR A CDR-1, CDR-2, CDR-3, and variable region sequences listed above.

[0085] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise (a) an alpha chain having the amino acid sequence set forth in SEQ ID NO: 14, or the amino acid sequence set forth in SEQ ID NO: 14 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions); and (b) a beta chain having the amino acid sequence set forth in SEQ ID NO: 22, or the amino acid sequence set forth in SEQ ID NO: 22 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). For example, a TCR or antigen-binding fragment thereof provided herein can (a) have the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2; (b) can comprise an alpha chain having the amino acid sequence set forth in SEQ ID NO: 14 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), with the proviso that the alpha chain includes the amino acid sequences set forth in SEQ ID NOs: 11, 12, and 13; and (c) a beta chain having the amino acid sequence set forth in SEQ ID NO: 22 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), with the proviso that the beta chain includes the amino acid sequences set forth in SEQ ID NOs: 19, 20, and 21. The CDR-1, CDR-2, CDR-3, and variable region sequences of any of TCRs B to P as identified in Table 1 may be substituted for the TCR A CDR-1, CDR-2, CDR-3, and variable region sequences listed above.

[0086] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein that have the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2 comprise a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 65 (or a variant of SEQ ID NO: 65 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 66 (or a variant of SEQ ID NO: 66 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 67 (or a variant of SEQ ID NO: 67 with one or two amino acid modifications). and a beta chain (or delta chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 73 (or a variant of SEQ ID NO: 73 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 74 (or a variant of SEQ ID NO: 74 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 75 (or a variant of SEQ ID NO: 75 with one or two amino acid modifications). Examples of such TCRs having these CDRs in the context of an MHC molecule and capable of binding to a peptide epitope of the minor histocompatibility antigen HA-2 include, but are not limited to, TCR D.

[0087] In some cases, an alpha chain (or gamma chain) having the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2 and having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 65 (or a variant of SEQ ID NO: 65 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 66 (or a variant of SEQ ID NO: 66 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 67 (or a variant of SEQ ID NO: 67 with one or two amino acid modifications). ), and a beta chain (or delta chain) having CDR-1 having the amino acid sequence set forth in SEQ ID NO: 73 (or a variant of SEQ ID NO: 73 with one or two amino acid modifications), CDR-2 having the amino acid sequence set forth in SEQ ID NO: 74 (or a variant of SEQ ID NO: 74 with one or two amino acid modifications), and CDR-3 having the amino acid sequence set forth in SEQ ID NO: 75 (or a variant of SEQ ID NO: 75 with one or two amino acid modifications), may comprise any suitable framework region.For example, such a TCR or antigen-binding fragment thereof may have a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO: 68 upstream of the amino acid sequence of SEQ ID NO: 65 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications), a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 68 between the amino acid sequences of SEQ ID NO: 65 and SEQ ID NO: 66 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications). a framework region 3 having the entire amino acid sequence set forth in SEQ ID NO: 68 located between the amino acid sequences of SEQ ID NO: 66 and SEQ ID NO: 67 (or variants of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications); and a framework region 4 having the entire amino acid sequence set forth in SEQ ID NO: 68 located downstream of the amino acid sequence of SEQ ID NO: 67 (or variants of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications). an alpha chain comprising: a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO: 76 upstream of the amino acid sequence of SEQ ID NO: 73 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications); a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 76 between the amino acid sequences of SEQ ID NO: 73 and SEQ ID NO: 74 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications); and a beta strand comprising a framework region 3 having the entire amino acid sequence set forth in SEQ ID NO: 76 between the amino acid sequence of SEQ ID NO: 75 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), and a framework region 4 having the entire amino acid sequence set forth in SEQ ID NO: 76 downstream of the amino acid sequence of SEQ ID NO: 75 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications).

[0088] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise an alpha chain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 68, and a beta chain comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 76. For example, a TCR or antigen-binding fragment thereof provided herein can comprise an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 68, and a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 76. In some cases, a TCR or antigen-binding fragment thereof provided herein can comprise (a) an alpha chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 68, and (b) a beta chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 76.

[0089] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise: (a) an alpha chain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 68 (with the proviso that the alpha chain comprises the amino acid sequences set forth in SEQ ID NOs: 65, 66, and 67); and (b) a beta chain comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 76 (with the proviso that the beta chain comprises the amino acid sequences set forth in SEQ ID NOs: 73, 74, and 75). For example, a TCR or antigen-binding fragment thereof provided herein can include (a) an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 68 (with the proviso that the alpha chain comprises the amino acid sequence set forth in SEQ ID NOs: 65, 66, and 67); and (b) a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity to the amino acid sequence set forth in SEQ ID NO: 76 (with the proviso that the beta chain comprises the amino acid sequence set forth in SEQ ID NOs: 73, 74, and 75).

[0090] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise (a) an alpha chain having the amino acid sequence set forth in SEQ ID NO: 68, or the amino acid sequence set forth in SEQ ID NO: 68 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions); and (b) a beta chain having the amino acid sequence set forth in SEQ ID NO: 76, or the amino acid sequence set forth in SEQ ID NO: 76 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). For example, a TCR or antigen-binding fragment thereof provided herein can (a) have the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2; (b) can comprise an alpha chain having the amino acid sequence set forth in SEQ ID NO: 68 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), with the proviso that the alpha chain includes the amino acid sequences set forth in SEQ ID NOs: 65, 66, and 67; and (c) a beta chain having the amino acid sequence set forth in SEQ ID NO: 76 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), with the proviso that the beta chain includes the amino acid sequences set forth in SEQ ID NOs: 73, 74, and 75.

[0091] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein that have the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule comprise a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 137 (or a variant of SEQ ID NO: 137 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 138 (or a variant of SEQ ID NO: 138 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 139 (or a variant of SEQ ID NO: 139 with one or two amino acid modifications). and a beta chain (or delta chain) having an amino acid sequence set forth in SEQ ID NO: 147 (or a variant of SEQ ID NO: 147 with one or two amino acid modifications). Examples of such TCRs having these CDRs in the context of an MHC molecule and capable of binding to a peptide epitope of the minor histocompatibility antigen HA-2 include, but are not limited to, TCR D.

[0092] In some cases, an alpha chain (or gamma chain) having the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2, and having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 137 (or a variant of SEQ ID NO: 137 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 138 (or a variant of SEQ ID NO: 138 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 139 (or a variant of SEQ ID NO: 139 with one or two amino acid modifications). ), and a beta chain (or delta chain) having CDR-1 having the amino acid sequence set forth in SEQ ID NO: 145 (or a variant of SEQ ID NO: 145 with one or two amino acid modifications), CDR-2 having the amino acid sequence set forth in SEQ ID NO: 146 (or a variant of SEQ ID NO: 146 with one or two amino acid modifications), and CDR-3 having the amino acid sequence set forth in SEQ ID NO: 147 (or a variant of SEQ ID NO: 147 with one or two amino acid modifications), may comprise any suitable framework region.For example, such a TCR or antigen-binding fragment thereof may have a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO: 140 upstream of the amino acid sequence of SEQ ID NO: 137 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications), a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 140 between the amino acid sequences of SEQ ID NO: 137 and SEQ ID NO: 138 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications). a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 140 located between the amino acid sequences of SEQ ID NO: 138 and SEQ ID NO: 139 (or variants of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications); and a framework region 4 having the entire amino acid sequence set forth in SEQ ID NO: 140 located downstream of the amino acid sequence of SEQ ID NO: 139 (or variants of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid modifications). an alpha chain comprising: a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO: 148 upstream of the amino acid sequence of SEQ ID NO: 145 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications); a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 148 between the amino acid sequences of SEQ ID NO: 145 and SEQ ID NO: 146 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications); a beta strand comprising a framework region 3 having the entire amino acid sequence set forth in SEQ ID NO: 148 between the amino acid sequences of SEQ ID NO: 6 and SEQ ID NO: 147 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), and a framework region 4 having the entire amino acid sequence set forth in SEQ ID NO: 148 downstream of the amino acid sequence of SEQ ID NO: 147 (or a variant of that sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications).

[0093] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise an alpha chain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 140, and a beta chain comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 148. For example, a TCR or antigen-binding fragment thereof provided herein can comprise an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 140, and a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 148. In some cases, a TCR or antigen-binding fragment thereof provided herein can comprise (a) an alpha chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 140, and (b) a beta chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 148.

[0094] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise: (a) an alpha chain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 140, with the proviso that the alpha chain comprises the amino acid sequences set forth in SEQ ID NOs: 137, 138, and 139; and (b) a beta chain comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 148, with the proviso that the beta chain comprises the amino acid sequences set forth in SEQ ID NOs: 145, 146, and 147. For example, a TCR or antigen-binding fragment thereof provided herein can include (a) an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 140 (with the proviso that the alpha chain comprises the amino acid sequence set forth in SEQ ID NOs: 137, 138, and 139); and (b) a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity to the amino acid sequence set forth in SEQ ID NO: 148 (with the proviso that the beta chain comprises the amino acid sequence set forth in SEQ ID NOs: 145, 146, and 147).

[0095] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise (a) an alpha chain having the amino acid sequence set forth in SEQ ID NO: 140, or the amino acid sequence set forth in SEQ ID NO: 140 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions); and (b) a beta chain having the amino acid sequence set forth in SEQ ID NO: 148, or the amino acid sequence set forth in SEQ ID NO: 148 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). For example, a TCR or antigen-binding fragment thereof provided herein (a) can have the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2; (b) can comprise an alpha chain having the amino acid sequence set forth in SEQ ID NO: 140 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), with the proviso that the alpha chain includes the amino acid sequences set forth in SEQ ID NOs: 137, 138, and 139; and (c) a beta chain having the amino acid sequence set forth in SEQ ID NO: 148 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), with the proviso that the beta chain includes the amino acid sequences set forth in SEQ ID NOs: 145, 146, and 147.

[0096] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein that have the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule may comprise an alpha chain (or gamma chain) and a beta chain (or delta chain) with the CDR sequences set forth in any of the TCRs in Table 1 (alpha or gamma chain CDR-1, CDR-2, and CDR-3, and delta chain beta CDR-1, CDR-2, and CDR-3). In some embodiments, the TCRs or antigen-binding fragments thereof provided herein that have the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise an alpha chain (or gamma chain) and a beta chain (or delta chain) having the CDR sequences (alpha or gamma chain CDR-1, CDR-2, and CDR-3, and delta chain beta CDR-1, CDR-2, and CDR-3) set forth in any of the TCRs in Table 1, wherein any one or more of the CDRs can independently be variant CDRs having one or two amino acid modifications from the CDR sequences listed in Table 1. Exemplary TCRs that have the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of these CDRs and an MHC molecule include, but are not limited to, TCR A, TCR B, TRC C, TRC D, TRC E, TRC F, TRC G, TRC H, TRC I, TRC J, TRC K, TRC L, TRC M, TRC N, TRC O, and TRC P.As an example, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule may comprise a CDR-1 having the amino acid sequence set forth in SEQ ID NO:29 (or a variant of SEQ ID NO:29 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO:30 (or a variant of SEQ ID NO:30 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO:31 (or a variant of SEQ ID NO:31 with one or two amino acid modifications). and a beta chain (or delta chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 37 (or a variant of SEQ ID NO: 37 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 38 (or a variant of SEQ ID NO: 38 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 39 (or a variant of SEQ ID NO: 39 with one or two amino acid modifications).

[0097] In some cases, the TCRs or antigen-binding fragments thereof provided herein having an alpha chain (or gamma chain) and a beta chain (or delta chain) with the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2 and having CDR sequences (alpha or gamma chain CDR-1, CDR-2, and CDR-3, and delta chain beta CDR-1, CDR-2, and CDR-3) as set forth in any of the TCRs in Table 1 may comprise any suitable framework region. In some cases, the TCRs or antigen-binding fragments thereof provided herein have the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2 and have an alpha chain (or gamma chain) and a beta chain (or delta chain) with CDR sequences as set forth in any of the TCRs in Table 1 (alpha or gamma chain CDR-1, CDR-2, and CDR-3, and delta chain beta CDR-1, CDR-2, and CDR-3), wherein any one or more of the CDRs can independently be variant CDRs having one or two amino acid modifications from the listed CDR sequences in Table 1. The TCRs or antigen-binding fragments thereof can comprise any suitable framework region (i.e., alpha chain and beta chain framework 1, 2, 3, and 4 regions). Alpha chain framework region 1 may have the amino acid sequence of any of the alpha variable (V) region amino acid sequences of any of the TCRs described in Table 1 upstream of the corresponding CDR-1 sequence, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications. Alpha chain framework region 2 may have the amino acid sequence of any of the alpha variable (V) region amino acid sequences of any of the TCRs described in Table 1 between the corresponding CDR-1 and CDR-2 sequences, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications.Alpha chain framework region 3 may have the amino acid sequence of any of the alpha variable (V) region amino acid sequences of any of the TCRs described in Table 1, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications, located between the corresponding CDR-2 and CDR-3 sequences. Alpha chain framework region 4 may have the amino acid sequence of any of the alpha variable (V) region amino acid sequences of any of the TCRs described in Table 1, downstream of the corresponding CDR-3 sequence, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications. Beta chain framework region 1 may have the amino acid sequence of any of the beta variable (V) region amino acid sequences of any of the TCRs described in Table 1, downstream of the corresponding CDR-1 sequence, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications. Beta chain framework region 2 may have the amino acid sequence of any of the beta variable (V) region amino acid sequences of any of the TCRs described in Table 1, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications between the corresponding CDR-1 and CDR-2 sequences. Beta chain framework region 3 may have the amino acid sequence of any of the beta variable (V) region amino acid sequences of any of the TCRs described in Table 1, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications between the corresponding CDR-2 and CDR-3 sequences. Beta chain framework region 4 may have the amino acid sequence of any of the beta variable (V) region amino acid sequences of any of the TCRs described in Table 1, or a variant of that sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications downstream of the corresponding CDR-3 sequence. The TCR or antigen-binding fragment thereof may have alpha chain framework regions 1, 2, 3, and 4 and the corresponding beta chain framework regions 1, 2, 3, and 4 of any of the TCRs described in Table 1.As an example, a TCE may have alpha chain framework regions 1, 2, 3, and 4 of SEQ ID NO:32 and beta chain framework regions 1, 2, 3, and 4 of SEQ ID NO:40.

[0098] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise an alpha chain comprising an amino acid sequence at least 90 percent identical to the amino acid sequence of an alpha chain variable (V) region of any of the TCRs described in Table 1, and a beta chain comprising an amino acid sequence at least 90% identical to the amino acid sequence of a corresponding beta chain variable (V) region described in Table 1. A TCR or antigen-binding fragment thereof provided herein can comprise an alpha chain comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identical to the amino acid sequence of an alpha chain variable (V) region of any of the TCRs described in Table 1, and a beta chain comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identical to the amino acid sequence of a corresponding beta chain variable (V) region described in Table 1. In some cases, a TCR or antigen-binding fragment thereof provided herein can comprise (a) an alpha chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence of an alpha chain variable (V) region of any of the TCRs set forth in Table 1, and (b) a beta chain comprising an amino acid sequence having 100 percent identity to the amino acid sequence of a corresponding beta chain variable (V) region set forth in Table 1. As an example, a TCR or antigen-binding fragment thereof provided herein can comprise an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 percent identity to the amino acid sequence of SEQ ID NO:32, and a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 percent identity to the amino acid sequence of SEQ ID NO:40.

[0099] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise: (a) an alpha chain comprising an amino acid sequence at least 90% identical to the amino acid sequence of an alpha chain variable (V) region of any of the TCRs set forth in Table 1, provided that the alpha chain comprises CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding alpha chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1; and (b) a beta chain comprising an amino acid sequence at least 90% identical to the amino acid sequence of a corresponding beta chain variable (V) region set forth in Table 1, provided that the beta chain comprises CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding beta chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1. The TCRs or antigen-binding fragments thereof provided herein comprise: (a) an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence of the alpha chain variable (V) region of any of the TCRs set forth in Table 1 (provided that the alpha chain comprises CDR-1, CDR-2, and CDR-3 sequences that are 100% identical to the corresponding alpha chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1); and (b) a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence of the corresponding beta chain variable (V) region set forth in Table 1, provided that the beta chain comprises CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding beta chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1.As an example, a TCR or antigen-binding fragment thereof provided herein can include an alpha chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence of SEQ ID NO: 32 (with the proviso that the alpha chain comprises the CDR-1, CDR-2, and CDR-3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively), and a beta chain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity to the amino acid sequence of SEQ ID NO: 40 (with the proviso that the beta chain comprises the CDR-1, CDR-2, and CDR-3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively).

[0100] In some cases, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise (a) an alpha chain having the amino acid sequence of an alpha chain variable (V) region of any of the TCRs described in Table 1, or amino acids of an alpha chain variable (V) region of any of the TCRs described in Table 1 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions); and (b) a beta chain having the amino acid sequence of a corresponding beta chain variable (V) region of any of the TCRs described in Table 1, or amino acids of a corresponding beta chain variable (V) region of any of the TCRs described in Table 1 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). The TCRs or antigen-binding fragments thereof provided herein can have (a) the ability to bind, in the context of an MHC molecule, to a peptide epitope of the minor histocompatibility antigen HA-2; and (b) an alpha chain having an amino acid sequence set forth in SEQ ID NO: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 with 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the alpha chain has 100% identity with the corresponding alpha chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1. (c) a beta chain having an amino acid sequence set forth in SEQ ID NO: 22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292, respectively, with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the beta chain comprises CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding beta chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1.As an example, a TCR or antigen-binding fragment thereof provided herein that has the ability to bind to a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule can comprise an alpha chain having the amino acid sequence set forth in SEQ ID NO: 32 with 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), whereby the alpha chain comprises the CDR-1, CDR-2, and CDR-3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively, and a beta chain having the amino acid sequence set forth in SEQ ID NO: 40 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), whereby the beta chain comprises the CDR-1, CDR-2, and CDR-3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively.

[0101] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein comprise: (a) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 11, 12, and 13, respectively; (b) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 29, 30, and 31, respectively; (c) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 47, 48, and 49, respectively; (d) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 65, 66, and 67, respectively; (e) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 83, 84, and 85, respectively; (f) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 101, 102, and 103, respectively; (g) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 119, 120, and 121, respectively; (h) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 137, 138, and 139, respectively; (i) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 155, 156, and 157, respectively; (j) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 173, 174, and 175, respectively; (k) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 191, 192, and 193, respectively; (l) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 209, 210, and 211, respectively; (m) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 227, 228, and 229, respectively; (n) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 245, 246, and 247, respectively; (o) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 263, 264, and 265, respectively; or (p) Contains a Vα or Vγ region containing CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 281, 282, and 283, respectively. Additionally, some of the TCRs provided have sequences that are at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences.

[0102] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein comprise: (a) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 19, 20, and 21, respectively; (b) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 37, 38, and 39, respectively; (c) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 55, 56, and 57, respectively; (d) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 73, 74, and 75, respectively; (e) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 91, 92, and 93, respectively; (f) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 109, 110, and 111, respectively; (g) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 127, 128, and 129, respectively; (h) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 145, 146, and 147, respectively; (i) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 163, 164, and 165, respectively; (j) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 181, 182, and 183, respectively; (k) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 199, 200, and 201, respectively; (l) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 217, 218, and 219, respectively; (m) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 235, 236, and 237, respectively; (n) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 253, 254, and 255, respectively; (o) CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 271, 272, and 273, respectively; (p) Contains a Vβ or Vδ region containing CDR-1, CDR-2, and CDR-3 comprising SEQ ID NOs: 289, 290, and 291, respectively. Additionally, some of the TCRs provided have sequences that are at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences.

[0103] In some embodiments, a TCR or antigen-binding fragment thereof provided herein comprises a Vα or Vγ region containing CDR-1, CDR-2, and CDR-3, each of which comprises the CDR-1, CDR-2, and CDR-3 amino acid sequences set forth in Table 1, such as in each row therein, and a Vβ or Vδ region containing CDR-1, CDR-2, and CDR-3, each of which comprises the CDR-1, CDR-2, and CDR-3 amino acid sequences set forth in Table 1, such as in each row therein. In some embodiments, a TCR or antigen-binding fragment thereof provided herein comprises a Vα region or Vγ region containing CDR-1, CDR-2, and CDR-3, which contain the CDR-1, CDR-2, and CDR-3 amino acid sequences contained within the Vα region or Vγ region amino acid sequence, respectively, as set forth in Table 1, such as in each row therein, and a Vβ region or Vδ region containing CDR-1, CDR-2, and CDR-3, which contain the CDR-1, CDR-2, and CDR-3 amino acid sequences contained within the Vα region or Vγ region amino acid sequence, respectively, as set forth in Table 1, such as in each row therein. In some embodiments, a TCR or antigen-binding fragment thereof provided herein comprises a Vα region or Vγ region amino acid sequence and the corresponding Vα region or Vγ region amino acid sequence as set forth in Table 1, such as in each row therein. Also provided are TCRs that contain sequences that are at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences. Exemplary TCRs containing such CDRs, or modified versions thereof described elsewhere herein, are also set forth in Table 1, such as in each row therein.

[0104] [Table 1]

[0105] In some examples, the TCRs or antigen-binding fragments thereof provided herein comprise a set of three CDRs (e.g., CDR-1, CDR-2, and CDR-3) set forth in Table 1 (e.g., SEQ ID NOs: 11-13; SEQ ID NOs: 29-31; SEQ ID NOs: 47-49; SEQ ID NOs: 65-67; ​​SEQ ID NOs: 83-85; SEQ ID NOs: 101-103; SEQ ID NOs: 119-121; SEQ ID NOs: 137-139; SEQ ID NOs: 155-157; SEQ ID NOs: 173-175; SEQ ID NOs: 191-193; SEQ ID NOs: 209-211; SEQ ID NOs: 227-229; SEQ ID NOs: 245-247; SEQ ID NOs: 263-265; or SEQ ID NOs: 281-283). and a beta chain (or delta chain) comprising a set of three CDRs (e.g., CDR-1, CDR-2, and CDR-3) set forth in Table 1 (e.g., SEQ ID NOs: 19-21; SEQ ID NOs: 37-39; SEQ ID NOs: 55-57; SEQ ID NOs: 73-75; SEQ ID NOs: 91-93; SEQ ID NOs: 109-111; SEQ ID NOs: 127-129; SEQ ID NOs: 145-147; SEQ ID NOs: 163-165; SEQ ID NOs: 181-183; SEQ ID NOs: 199-201; SEQ ID NOs: 217-219; SEQ ID NOs: 235-237; SEQ ID NOs: 253-255; SEQ ID NOs: 271-273; or SEQ ID NOs: 289-291).

[0106] In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO: 11, CDR-2 comprising SEQ ID NO: 12, and CDR-3 comprising SEQ ID NO: 12, and the Vβ region comprises CDR-1 comprising SEQ ID NO: 19, CDR-2 comprising SEQ ID NO: 20, and CDR-3 comprising SEQ ID NO: 21. In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO: 29, CDR-2 comprising SEQ ID NO: 30, and CDR-3 comprising SEQ ID NO: 31, and the Vβ region comprises CDR-1 comprising SEQ ID NO: 37, CDR-2 comprising SEQ ID NO: 38, and CDR-3 comprising SEQ ID NO: 39. In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO: 47, CDR-2 comprising SEQ ID NO: 48, and CDR-3 comprising SEQ ID NO: 49, and the Vβ region comprises CDR-1 comprising SEQ ID NO: 55, CDR-2 comprising SEQ ID NO: 56, and CDR-3 comprising SEQ ID NO: 57. In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO:65, CDR-2 comprising SEQ ID NO:66, and CDR-3 comprising SEQ ID NO:67, and the Vβ region comprises CDR-1 comprising SEQ ID NO:73, CDR-2 comprising SEQ ID NO:74, and CDR-3 comprising SEQ ID NO:75. In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO:83, CDR-2 comprising SEQ ID NO:84, and CDR-3 comprising SEQ ID NO:85, and the Vβ region comprises CDR-1 comprising SEQ ID NO:91, CDR-2 comprising SEQ ID NO:92, and CDR-3 comprising SEQ ID NO:93. In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO:101, CDR-2 comprising SEQ ID NO:102, and CDR-3 comprising SEQ ID NO:103, and the Vβ region comprises CDR-1 comprising SEQ ID NO:109, CDR-2 comprising SEQ ID NO:110, and CDR-3 comprising SEQ ID NO:111. In some embodiments, the Vα region comprises a CDR-1 comprising SEQ ID NO: 119, a CDR-2 comprising SEQ ID NO: 120, and a CDR-3 comprising SEQ ID NO: 121, and the Vβ region comprises a CDR-1 comprising SEQ ID NO: 127, a CDR-2 comprising SEQ ID NO: 128, and a CDR-3 comprising SEQ ID NO: 129.In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO: 137, CDR-2 comprising SEQ ID NO: 138, and CDR-3 comprising SEQ ID NO: 139, and the Vβ region comprises CDR-1 comprising SEQ ID NO: 145, CDR-2 comprising SEQ ID NO: 146, and CDR-3 comprising SEQ ID NO: 147. In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO: 155, CDR-2 comprising SEQ ID NO: 156, and CDR-3 comprising SEQ ID NO: 157, and the Vβ region comprises CDR-1 comprising SEQ ID NO: 163, CDR-2 comprising SEQ ID NO: 164, and CDR-3 comprising SEQ ID NO: 165. In some embodiments, the Vα region comprises CDR-1 comprising SEQ ID NO: 173, CDR-2 comprising SEQ ID NO: 174, and CDR-3 comprising SEQ ID NO: 175, and the Vβ region comprises CDR-1 comprising SEQ ID NO: 181, CDR-2 comprising SEQ ID NO: 182, and CDR-3 comprising SEQ ID NO: 183. In some embodiments, the Vα region comprises a CDR-1 comprising SEQ ID NO: 191, a CDR-2 comprising SEQ ID NO: 192, and a CDR-3 comprising SEQ ID NO: 193, and the Vβ region comprises a CDR-1 comprising SEQ ID NO: 199, a CDR-2 comprising SEQ ID NO: 200, and a CDR-3 comprising SEQ ID NO: 201. In some embodiments, the Vα region comprises a CDR-1 comprising SEQ ID NO: 209, a CDR-2 comprising SEQ ID NO: 210, and a CDR-3 comprising SEQ ID NO: 211, and the Vβ region comprises a CDR-1 comprising SEQ ID NO: 217, a CDR-2 comprising SEQ ID NO: 218, and a CDR-3 comprising SEQ ID NO: 219. In some embodiments, the Vα region comprises a CDR-1 comprising SEQ ID NO: 227, a CDR-2 comprising SEQ ID NO: 228, and a CDR-3 comprising SEQ ID NO: 229, and the Vβ region comprises a CDR-1 comprising SEQ ID NO: 235, a CDR-2 comprising SEQ ID NO: 236, and a CDR-3 comprising SEQ ID NO: 237. In some embodiments, the Vα region comprises a CDR-1 comprising SEQ ID NO: 245, a CDR-2 comprising SEQ ID NO: 246, and a CDR-3 comprising SEQ ID NO: 247, and the Vβ region comprises a CDR-1 comprising SEQ ID NO: 253, a CDR-2 comprising SEQ ID NO: 254, and a CDR-3 comprising SEQ ID NO: 255.In some embodiments, the Vα region comprises a CDR-1 comprising SEQ ID NO: 263, a CDR-2 comprising SEQ ID NO: 264, and a CDR-3 comprising SEQ ID NO: 265, and the Vβ region comprises a CDR-1 comprising SEQ ID NO: 271, a CDR-2 comprising SEQ ID NO: 272, and a CDR-3 comprising SEQ ID NO: 273. In some embodiments, the Vα region comprises a CDR-1 comprising SEQ ID NO: 281, a CDR-2 comprising SEQ ID NO: 282, and a CDR-3 comprising SEQ ID NO: 283, and the Vβ region comprises a CDR-1 comprising SEQ ID NO: 289, a CDR-2 comprising SEQ ID NO: 290, and a CDR-3 comprising SEQ ID NO: 291.

[0107] In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region of SEQ ID NO: 14, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 22. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 32, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 40. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 50, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 58. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region of SEQ ID NO:68, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO:76. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO:86, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO:94. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO:104, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO:112. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 122, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 130. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 140, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 148.In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 158, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 166. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 176, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 184. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 194, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 202. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 212, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 220. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 230, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 238. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 248, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 256. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 266, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 274. In some embodiments, the Vα region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vα region sequence of SEQ ID NO: 284, and the Vβ region comprises CDR-1, CDR-2, and CDR-3 as contained in the Vβ region sequence of SEQ ID NO: 292.

[0108] In some embodiments, the Vα region comprises SEQ ID NO: 14, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 22, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 32, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 40, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 50, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 58, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 68, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 76, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO:86, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO:94, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO:104, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO:112, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO:122, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO:130, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO:140, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO:148, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 158, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 166, or a sequence having at least 90% sequence identity thereto.In some embodiments, the Vα region comprises SEQ ID NO: 176, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 184, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 194, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 202, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 212, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 220, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 230, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 238, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 248, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 256, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 266, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 274, or a sequence having at least 90% sequence identity thereto. In some embodiments, the Vα region comprises SEQ ID NO: 284, or a sequence having at least 90% sequence identity thereto, and the Vβ region comprises SEQ ID NO: 292, or a sequence having at least 90% sequence identity thereto.

[0109] In some embodiments, the Vα region comprises SEQ ID NO: 14, and the Vβ region comprises SEQ ID NO: 22. In some embodiments, the Vα region comprises SEQ ID NO: 32, and the Vβ region comprises SEQ ID NO: 40. In some embodiments, the Vα region comprises SEQ ID NO: 50, and the Vβ region comprises SEQ ID NO: 58. In some embodiments, the Vα region comprises SEQ ID NO: 68, and the Vβ region comprises SEQ ID NO: 76. In some embodiments, the Vα region comprises SEQ ID NO: 86, and the Vβ region comprises SEQ ID NO: 94. In some embodiments, the Vα region comprises SEQ ID NO: 104, and the Vβ region comprises SEQ ID NO: 112. In some embodiments, the Vα region comprises SEQ ID NO: 122, and the Vβ region comprises SEQ ID NO: 130. In some embodiments, the Vα region comprises SEQ ID NO: 140, and the Vβ region comprises SEQ ID NO: 148. In some embodiments, the Vα region comprises SEQ ID NO: 158, and the Vβ region comprises SEQ ID NO: 166. In some embodiments, the Vα region comprises SEQ ID NO: 176, and the Vβ region comprises SEQ ID NO: 184. In some embodiments, the Vα region comprises SEQ ID NO: 194, and the Vβ region comprises SEQ ID NO: 202. In some embodiments, the Vα region comprises SEQ ID NO: 212, and the Vβ region comprises SEQ ID NO: 220. In some embodiments, the Vα region comprises SEQ ID NO: 230, and the Vβ region comprises SEQ ID NO: 238. In some embodiments, the Vα region comprises SEQ ID NO: 248, and the Vβ region comprises SEQ ID NO: 256. In some embodiments, the Vα region comprises SEQ ID NO: 266, and the Vβ region comprises SEQ ID NO: 274. In some embodiments, the Vα region comprises SEQ ID NO: 284, and the Vβ region comprises SEQ ID NO: 292.

[0110] D. Exemplary Constant Domains In some embodiments, the alpha chain of a TCR or antigen-binding fragment thereof provided herein further comprises an alpha constant (Cα) region or a portion thereof. In some aspects, the beta chain further comprises a beta constant (Cβ) region or a portion thereof. Thus, in some embodiments, a TCR provided herein (e.g., an anti-HA-2 TCR provided herein) or antigen-binding fragment thereof comprises an alpha chain comprising a Vα region and a Cα domain or a portion thereof, and / or a beta chain comprising a Vβ region and a Cβ domain or a portion thereof. In some embodiments, the gamma chain of a TCR or antigen-binding fragment thereof provided herein further comprises a gamma constant (Cγ) region or a portion thereof. In some aspects, the delta chain further comprises a delta constant (Cδ) region or a portion thereof. Thus, in some embodiments, a TCR provided herein (e.g., an anti-HA-2 TCR provided herein) or antigen-binding fragment thereof comprises a gamma chain comprising a Vγ region and a Cγ domain or a portion thereof, and / or a delta chain comprising a Vδ region and a Cδ domain or a portion thereof.

[0111] In some embodiments, the α and β chains, or the γ and δ chains, of the TCRs provided herein each further contain a constant domain. In some embodiments, the Cα and Cβ domains, or the Cγ and Cδ domains, are individually mammalian (e.g., human or mouse constant domains). In some embodiments, the constant domains are adjacent to the cell membrane. For example, in some cases, the extracellular portion of the TCR formed by the two chains contains two membrane-proximal constant domains and two membrane-distal variable domains, each of which contains a CDR.

[0112] In some aspects, provided herein are TCRs that contain human constant domains, such as an alpha chain containing a human Cα domain and a beta chain containing a human Cβ domain, or a gamma chain containing a human Cγ domain and a delta chain containing a human Cδ domain. In some embodiments, the provided TCRs are fully human. Among the TCRs provided are TCRs that contain human constant domains, e.g., fully human TCRs, whose expression and / or activity is unaffected or substantially unaffected by the presence of endogenous human TCRs, e.g., when expressed in human cells, e.g., human T cells, such as primary human T cells.

[0113] In some embodiments, each of the Cα and Cβ domains, or each of the Cγ and Cδ domains, is human. In some embodiments, Cα is encoded by the TRAC gene (IMGT nomenclature) or a variant thereof. In some embodiments, Cβ is encoded by the TRBC1 gene or the TRBC2 gene (IMGT nomenclature) or a variant thereof. In some embodiments, Cγ is encoded by the TRGC1 gene or the TRGC2 gene (IMGT nomenclature) or a variant thereof. In some embodiments, Cδ is encoded by the TRDC gene (IMGT nomenclature) or a variant thereof.

[0114] In some embodiments, the Cα domain or a variant thereof has or comprises a sequence of amino acids set forth in SEQ ID NO: 3 or 5, or a sequence of amino acids exhibiting at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 3 or 5. In some embodiments, the Cα domain has or comprises a sequence of amino acids set forth in SEQ ID NO: 3. In some embodiments, the Cα domain has or comprises a sequence of amino acids set forth in SEQ ID NO: 5. In some embodiments, the Cβ domain or a variant thereof has or comprises the sequence of amino acids set forth in SEQ ID NO: 7 or 9, or a sequence of amino acids exhibiting at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 7 or 9. In some embodiments, the Cβ domain has or comprises the sequence of amino acids set forth in SEQ ID NO: 7. In some embodiments, the Cβ domain has or comprises the sequence of amino acids set forth in SEQ ID NO: 9. In some embodiments, the TCR comprises the Cα domain and the Cβ domain set forth in SEQ ID NO: 3 and 7, respectively. In some embodiments, the TCR comprises the Cα domain and the Cβ domain set forth in SEQ ID NO: 5 and 7, respectively. In some embodiments, the TCR comprises the Cα domain and the Cβ domain set forth in SEQ ID NO: 3 and 9, respectively. In some embodiments, the TCR comprises the Cα and Cβ domains set forth in SEQ ID NOs: 5 and 9, respectively.

[0115] In some embodiments, the Cγ domain or a variant thereof has or comprises a sequence of amino acids set forth in SEQ ID NO: 356 or 358, or a sequence of amino acids exhibiting at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 311 or 312. In some embodiments, the Cγ domain has or comprises a sequence of amino acids set forth in SEQ ID NO: 311. In some embodiments, the Cγ domain has or comprises a sequence of amino acids set forth in SEQ ID NO: 312. In some embodiments, the Cδ domain or variant thereof has or comprises a sequence of amino acids set forth in SEQ ID NO: 313, or a sequence of amino acids exhibiting at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 313. In some embodiments, the TCR comprises a Cγ domain and a Cδ domain set forth in SEQ ID NOs: 311 and 313, respectively. In some embodiments, the TCR comprises a Cγ domain and a Cδ domain set forth in SEQ ID NOs: 312 and 313, respectively.

[0116] In some embodiments, variants of the Cα domain contain at least one non-native cysteine ​​replacement, such as any of the replacements described herein. In some embodiments, variants of the Cβ domain contain at least one non-native cysteine ​​replacement, such as any of the replacements described herein.

[0117] In some embodiments, any of the provided TCRs or antigen-binding fragments thereof can be human / mouse chimeric TCRs. In some cases, the TCRs or antigen-binding fragments thereof provided herein comprise an alpha chain and / or beta chain, or a gamma chain and / or delta chain, comprising a mouse constant domain. In some embodiments, the Cα domain and / or Cβ domain, or the Cγ domain and / or Cδ domain, is a mouse Cα domain and / or mouse Cβ domain, or a mouse Cγ domain and / or mouse Cδ domain. In some embodiments, the Cα domain and / or Cβ domain, or the Cγ domain and / or Cδ domain is or comprises any of the Cα domain and / or Cβ domain, or the Cγ domain and / or Cδ domain described in WO 2015 / 184228, WO 2015 / 009604, and WO 2015 / 009606.

[0118] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein comprise variants of the alpha and / or beta chain, or the gamma and / or delta chain. In some embodiments, the variants comprise the amino acid sequences of any of the TCRs described herein, with one, two, three, or more amino acid substitutions in the constant domains of the alpha or beta chain. In some embodiments, the TCRs (or functional portions thereof) comprising the substituted amino acid sequences advantageously provide one or more of reduced mispairing with endogenous TCR chains, increased expression by host cells, increased recognition of the HA-2 target, and increased anti-tumor activity compared to the parent TCR comprising the unsubstituted amino acid sequence.

[0119] In some embodiments, the constant domain contains substituted amino acid sequences of mouse constant domains of the TCR α and β chains or TCR γ and δ chains corresponding to all or part of the unsubstituted mouse Cα and Cβ domains or the mouse Cγ and Cδ domains. In some embodiments, the TCR may be a heterodimer of α and β chains or γ and δ chains linked by one disulfide bond or multiple disulfide bonds, etc. In some embodiments, the constant domain of the TCR may contain a short linking sequence in which cysteine ​​residues form disulfide bonds, thereby linking the two chains of the TCR. In some embodiments, the TCR may have additional cysteine ​​residues in each of the α and β chains or the γ and δ chains such that the TCR contains two disulfide bonds in the constant domain. In some embodiments, the constant domain and the variable domain each contain disulfide bonds formed by cysteine ​​residues.

[0120] In some embodiments, the TCRs provided herein may contain an introduced disulfide bond or multiple disulfide bonds. In some embodiments, native disulfide bonds are absent. In some embodiments, one or more of the native cysteines (e.g., in the α and β, or γ and δ, chain constant domains) that form native interchain disulfide bonds are substituted with another residue, such as serine or alanine. In some embodiments, the introduced disulfide bonds can be formed by mutating non-cysteine ​​residues on the alpha and beta chains, such as the α and β or γ and δ, chain constant domains, to cysteine. Opposing cysteines in the TCR α and β or TCR γ and δ chains link the TCR α and β, or TCR γ and δ, chain constant domains of the substituted TCRs to each other, providing a disulfide bond that is not present in TCRs comprising unsubstituted constant domains where native disulfide bonds exist, such as unsubstituted native human constant domains or unsubstituted native mouse constant domains. In some embodiments, the presence of non-native cysteine ​​residues in a recombinant TCR (e.g., resulting in one or more non-native disulfide bonds) may favor production of the desired recombinant TCR in a cell into which it is introduced over expression of a mismatched TCR pair containing native TCR chains.

[0121] Exemplary non-native disulfide bonds in TCRs are described in published International PCT Patent Applications Nos. 2006 / 000830 and 2006 / 037960. In some embodiments, cysteines may be introduced or substituted at residues corresponding to Thr48 of the Cα domain and Ser57 of the Cβ domain, Thr45 of the Cα domain and Ser77 of the Cβ domain, Tyr10 of the Cα domain and Ser17 of the Cβ domain, Thr45 of the Cα domain and Asp59 of the Cβ domain, and / or Ser15 of the Cα domain and Glu15 of the Cβ domain.

[0122] In some embodiments, any of the provided cysteine ​​mutations can be made at a corresponding position in another sequence, for example, the sequence of the human or mouse Cα and / or Cβ domains, or the Cγ and / or Cδ domains described above. The term "corresponding" with respect to a protein position, such as a statement that an amino acid position in a disclosed sequence, as set forth in the sequence listing, is a "corresponding" amino acid position, refers to an amino acid position identified upon alignment with the disclosed sequence based on a structural sequence alignment or using a standard alignment algorithm, such as the GAP algorithm. For example, corresponding residues can be determined by aligning a reference sequence with the Cα sequence set forth in either SEQ ID NO: 3 or 5, or the Cβ sequence set forth in SEQ ID NO: 7 or 300, using structural alignment methods as described herein. By aligning the sequences, corresponding residues can be identified, for example, using conserved and identical amino acid residues as a guide.

[0123] In some embodiments, the TCRs or antigen-binding fragments thereof provided herein are or have at least 90% sequence identity to the sequence of amino acids set forth in any of SEQ ID NOs: 17, 35, 53, 71, 89, 107, 125, 143, 161, 179, 197, 215, 233, 251, 269, and 287, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence. and / or a beta or delta chain that is or comprises a sequence of amino acids set forth in any of SEQ ID NOs: 25, 43, 61, 79, 97, 115, 133, 151, 169, 187, 205, 223, 241, 259, 277, and 295, or a sequence having at least 90% sequence identity thereto, such as a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to such a sequence.

[0124] Exemplary TCRs or antigen-binding fragments include those set forth in Table 2, such as in each row therein. In some embodiments, the Vα and Vβ regions, or the Vγ and Vδ regions, contain amino acid sequences corresponding to a SEQ ID NO: set forth in Table 2, such as in each row therein. In some embodiments, the Vα and Vβ regions, or the Vγ and Vδ regions, contain CDR-1, CDR-2, and CDR-3 sequences contained within the Vα and Vβ regions set forth in Table 2, such as in each row therein. In some aspects, the TCRs contain constant alpha and constant beta domain sequences, such as those corresponding to a SEQ ID NO: set forth in Table 2, such as in each row therein. In some cases, the TCRs contain complete sequences including variable and constant domains, such as sequences corresponding to a SEQ ID NO: set forth in Table 2 ("complete alpha-P2A-beta"), such as in each row therein. Each of the TCRs in Table 2 can also be provided in a beta-P2A-alpha format (i.e., the beta full chain amino acid sequence is at the amino terminus, followed by the P2A sequence, then the alpha full chain amino acid sequence; e.g., SEQ ID NO: 314, 316, or 318). Additionally, some of the TCRs provided contain sequences that are at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences. Exemplary TCRs containing such sequences, or modified versions thereof described elsewhere herein, are also each described in Table 2, e.g., in each row therein. In some aspects, the exemplary TCRs provided contain mature Vα and / or Vβ regions, or mature Vγ and / or Vδ regions, that, when expressed as mature proteins, are fully processed and expressed, e.g., do not include a signal sequence (e.g., from cleavage of the signal sequence).

[0125] [Table 2] *Alternatively, the sequence can also be constructed as beta-P2A-alpha.

[0126] In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 17, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 25, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 35, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 43, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 53, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 61, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 71, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 79, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:89, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:97, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:107, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:115, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:125, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:133, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:143, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:151, or a sequence having at least 90% sequence identity thereto.In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 161, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 169, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 179, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 187, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 197, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 205, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 215, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO: 223, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:233, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:241, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:251, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:259, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:269, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:277, or a sequence having at least 90% sequence identity thereto. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:287, or a sequence having at least 90% sequence identity thereto, and the beta or delta chain comprises SEQ ID NO:295, or a sequence having at least 90% sequence identity thereto.

[0127] In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 17, and the beta or delta chain comprises SEQ ID NO: 25. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 35, and the beta or delta chain comprises SEQ ID NO: 43. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 53, and the beta or delta chain comprises SEQ ID NO: 61. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 71, and the beta or delta chain comprises SEQ ID NO: 79. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 89, and the beta or delta chain comprises SEQ ID NO: 97. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 107, and the beta or delta chain comprises SEQ ID NO: 115. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 125, and the beta or delta chain comprises SEQ ID NO: 133. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 143, and the beta or delta chain comprises SEQ ID NO: 151. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 161, and the beta or delta chain comprises SEQ ID NO: 169. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 179, and the beta or delta chain comprises SEQ ID NO: 187. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 197, and the beta or delta chain comprises SEQ ID NO: 205. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 215, and the beta or delta chain comprises SEQ ID NO: 223. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 233, and the beta or delta chain comprises SEQ ID NO: 241. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 251, and the beta or delta chain comprises SEQ ID NO: 259. In some embodiments, the alpha or gamma chain comprises SEQ ID NO: 269, and the beta or delta chain comprises SEQ ID NO: 277. In some embodiments, the alpha or gamma chain comprises SEQ ID NO:287 and the beta or delta chain comprises SEQ ID NO:295.

[0128] In some embodiments, the TCR comprises an amino acid sequence of any of SEQ ID NOs: 27, 45, 63, 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, 297, 314, 316, and 318, or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any of SEQ ID NOs: 27, 45, 63, 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, 297, 314, 316, and 318.

[0129] E. Anti-HA2 TCR Single Chain Variable Fragment Any of the anti-HA2 TCR CDRs or variable regions described herein can be used in the formation of single chain variable fragments (scFvs), bispecific T cell engagers (BiTEs), chimeric T cell receptors (CARs), or other proteins containing scFvs, BiTEs, or CARs.

[0130] "scFv" refers to the variable heavy chain regions (V) of immunoglobulins linked by a short linker peptide. H ) and the variable light chain region (V L ), or a fusion protein comprising the variable alpha region (Vα) and variable beta region (Vβ) of a TCR. scFvs retain the antigen-binding properties of the intact immunoglobulin or TCR from which the variable regions are derived. Nucleic acids encoding anti-HA-2 scFvs are also contemplated.

[0131] In some embodiments, an anti-HA-2 scFv is described that comprises the Vα and Vβ regions of any of the described HA-2 TCRs, wherein the Vα and Vβ regions are linked via a linker peptide, and the anti-HA-2 scFv retains the antigen-binding properties of the HA-2 TCR from which the variable regions are derived. Nucleic acids encoding anti-HA-2 scFvs are also contemplated. The anti-HA2 scFv can be formed by linking the C-terminus of the Vα chain to the N-terminus of the Vβ chain. Alternatively, the C-terminus of the Vβ can be linked to the N-terminus of the Vα chain. The peptide linker can be from about 10 to about 25 amino acids. In some embodiments, the scFv peptide linker is glycine-rich. The scFv peptide linker can be (G4S) x where x can be, but is not limited to, an integer between 2 and 5 (inclusive). In some embodiments, the linked scFv peptide comprises Gly-Gly-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser (i.e., also referred to as [(Gly)Ser], (G4S) or G4S(x3)). In some embodiments, the scFv peptide linker consists of G4S(x3).

[0132] Bispecific T cell engagers (BiTEs) are recombinant molecules containing two flexibly linked antigen-binding domains, such as scFvs. In a typical BiTE, one antigen-binding domain of the BiTE is specific for CD3 or other activating antigen on immune cells, and the second antigen-binding domain has affinity for a second antigen, such as a tumor antigen or an antigen on target cells. BiTEs can be used to target T cells that contain the CD3 receptor along with target cells (as described in WO 99054440, WO 2005040220, and WO 2008119567). BiTEs can transiently bind T cells to target cells and simultaneously activate the cytolytic activity of the T cells.

[0133] In some embodiments, anti-HA-2 BiTE molecules are described that comprise a first antigen-binding domain (e.g., an scFv) specific for an activating antigen on an immune cell and a second antigen-binding domain comprising an HA-2 binding domain or scFv comprising the variable region or CDRs of any of the described TCRs. In some embodiments, the BiTE comprises an anti-CD3 scFv and an anti-HA-2 scFv comprising the Vα and Vβ regions of any of the described HA-2 TCRs. Nucleic acids encoding anti-HA-2 BiTEs are also contemplated.

[0134] A "chimeric antigen receptor" or "CAR" is a recombinant protein comprising an antigen binding domain (e.g., an antigen binding fragment of any of the described TCRs) linked via a transmembrane domain to a cell signaling and / or cell activation domain. The cell signaling domain can be, but is not limited to, a T cell signaling domain. When utilized in a CAR, the antigen binding fragment of the described TCR can be provided as an scFv. A CAR can be a first generation CAR T cell, a second generation CAR T cell, a third generation CAR T cell, a fourth generation CAR T cell, a dual antigen receptor CAR T cell, or a CAR with an inducible suicide gene. The CAR may be a T cell, a T cell, or a combination thereof. The CAR may have a single signaling and / or cell activation domain, multiple signaling and / or cell activation domains, or one or more signaling and / or cell activation domains and one or more costimulatory domains. The signaling and / or cell activation domain or costimulatory domain may be, but is not limited to, a CD3 zeta domain, a CD28 domain, a CD137 domain, an ICOS domain, a CD27 domain, an OX40 domain, an LFA1 domain, a PD-1 domain, a CD150 domain, a CD244 domain, an NKG2D domain, and a DAP10 domain.

[0135] In some embodiments, an anti-HA-2 CAR is described that comprises an antigen-binding fragment of any of the described HA-2 TCRs, or an scFv that comprises the variable region or CDRs, transmembrane domain, and signaling and / or cell activation domain of any of the described TCRs. In some embodiments, the antigen-binding fragment of any of the described HA-2 TCRs comprises an anti-HA-2 scFv, which comprises the Vα and Vβ regions of any of the described HA-2 TCRs. The anti-HA-2 CAR can be, but is not limited to, a first-generation CAR T cell, a second-generation CAR T cell, a third-generation CAR T cell, a fourth-generation CAR T cell, a dual antigen receptor CAR T cell, or a CAR T cell with an inducible suicide gene, or a combination thereof. Nucleic acids encoding the anti-HA-2 CAR are also contemplated.

[0136] II. Nucleic Acids Encoding TCRs Also provided herein are nucleic acids, such as polynucleotides or nucleic acid molecules, that encode any of the provided TCRs or antigen-binding fragments thereof. Nucleic acids can include those that contain natural and / or non-natural nucleotides and bases, including, for example, those with backbone modifications. The terms "nucleic acid molecule," "nucleic acid," and "polynucleotide" may be used interchangeably and refer to a polymer of nucleotides. Such polymers of nucleotides may contain natural and / or non-natural nucleotides and include, but are not limited to, DNA, RNA, and PNA. A "nucleic acid sequence" refers to a linear sequence of nucleotides that comprises a nucleic acid molecule or polynucleotide.

[0137] In some embodiments, the TCRs or antigen-binding portions thereof provided herein may be recombinantly produced native proteins or mutant forms thereof that have altered one or more properties, such as binding characteristics. In some aspects, the nucleic acid is synthetic. In some cases, the nucleic acid is or contains cDNA. In some aspects, the polynucleotide can be modified for use in the constructs described herein, such as for codon optimization. In some cases, the sequence can be designed to contain terminal restriction site sequences for purposes of cloning into a vector.

[0138] In some embodiments, the TCRs or antigen-binding portions thereof provided herein can be synthetically generated from knowledge of the sequence of the TCR.

[0139] In some embodiments, the polynucleotide contains a nucleic acid sequence encoding an alpha chain and / or a nucleotide sequence encoding a beta chain, hi some embodiments, the polynucleotide contains a nucleic acid sequence encoding a gamma chain and / or a nucleotide sequence encoding a delta chain.

[0140] In some embodiments, the nucleotide sequence encoding the alpha or gamma chain, and / or the nucleotide sequence encoding the beta or delta chain, or any domain, region, or portion thereof, is codon-optimized. Typically, codon optimization involves balancing the proportion of selected codons with the published abundance of human transfer RNA to avoid overloading or limiting codons. Most amino acids are encoded by more than one codon, and codon usage varies between organisms. Differences in codon usage between the transfected gene and the host cell can affect protein expression and the immunogenicity of the nucleic acid construct. Generally, for codon optimization, codons are selected to select codons that are balanced with human usage. Typically, the codon redundancy for an amino acid is such that different codons encode a single amino acid. In some embodiments, when selecting a codon for replacement, it may be desirable for the resulting mutation to be silent, so that the codon change does not affect the amino acid sequence. Generally, the last nucleotide of a codon can be changed without affecting the amino acid sequence.

[0141] In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 15, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 23, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 33, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 41, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 51, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 59, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 69, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 77, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO:87, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO:95, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO:105, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO:113, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO:123, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO:131, or a sequence having at least 90% sequence identity thereto.In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 141, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 149, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 159, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 167, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 177, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 185, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 195, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 203, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 213, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 221, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 231, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 239, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 249, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 257, or a sequence having at least 90% sequence identity thereto.In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 267, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 275, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 285, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 293, or a sequence having at least 90% sequence identity thereto.

[0142] In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 16, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 24, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 34, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 42, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 52, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 60, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 70, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 78, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 88, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 96, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 106, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 114, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 124, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 132, or a sequence having at least 90% sequence identity thereto.In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 142, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 150, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 160, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 168, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 178, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 186, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 196, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 204, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 214, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 222, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 232, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 240, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 250, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 258, or a sequence having at least 90% sequence identity thereto.In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 268, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 276, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Vα region comprises SEQ ID NO: 286, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ region comprises SEQ ID NO: 294, or a sequence having at least 90% sequence identity thereto.

[0143] In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 18, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 26, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 36, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 44, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 54, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 62, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:72, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:80, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:90, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:98, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:108, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:116, or a sequence having at least 90% sequence identity thereto.In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 126, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 134, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 144, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 152, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 162, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 170, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 180, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 188, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 198, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 206, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 216, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 224, or a sequence having at least 90% sequence identity thereto.In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:234, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:242, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:252, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:260, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:270, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:278, or a sequence having at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:288, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:296, or a sequence having at least 90% sequence identity thereto.

[0144] In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 18, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 26. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 36, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 44. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 54, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 62. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 72, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 80. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 90, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 98. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 108, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 116. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 126, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 134. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 144, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 152. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 162, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 170. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 180, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 188. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 198, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 206.In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:216, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:224. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:234, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:242. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:252, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:260. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:270, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:278. In some embodiments, the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:288, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:296. Also provided herein are nucleic acids or polynucleotides that contain sequences that are at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences. Also provided are embodiments of one or more chains (e.g., alpha or gamma and / or beta or delta chains) of a TCR or binding fragment thereof encoded by any of such polynucleotides.

[0145] In some embodiments, the nucleic acid sequence encoding the alpha or gamma chain comprises any of SEQ ID NOs: 15, 33, 51, 69, 87, 105, 123, 141, 159, 177, 195, 213, 231, 249, 267, and 285, degenerate sequences thereof, or sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto. In some embodiments, the nucleotide sequence encoding the beta or delta chain comprises any of SEQ ID NOs: 23, 41, 59, 77, 95, 113, 131, 149, 167, 185, 203, 221, 239, 257, 275, 293, degenerate sequences thereof, or sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto.

[0146] In some embodiments, the alpha or gamma and / or beta or delta chains of the TCR are encoded by a sequence of nucleotides that includes a signal peptide (also referred to as a leader sequence). Non-limiting examples of such signal peptides are signal peptides having or comprising the sequence of amino acids set forth in any of SEQ ID NOs: 299, 300, 301, 302, 303, 304, 305, 306, 307, and 308.

[0147] In some embodiments, the nucleic acid encoding the alpha or gamma chain and the nucleic acid encoding the beta or delta chain may be connected via a linker, such as those optionally described elsewhere herein.

[0148] In some embodiments, the nucleic acid encoding the alpha or gamma chain and the nucleic acid encoding the beta or delta chain can be connected via a cleavable linker sequence or a peptide that causes ribosome skipping (e.g., T2A or P2A), such as those optionally described elsewhere herein. The P2A amino acid sequence can be, but is not limited to, the sequence of SEQ ID NO: 309. The nucleic acid sequence encoding the P2A sequence can be, but is not limited to, the nucleic acid sequence of SEQ ID NO: 310, a nucleic acid sequence encoding a P2A peptide that is at least 90% identical to the amino acid sequence of SEQ ID NO: 309, or a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 309.

[0149] In some embodiments, the nucleic acid sequences encoding the alpha and beta TCR chains connected via a cleavable linker sequence have at least 90% identity to any of the nucleotide sequences of SEQ ID NOs: 28, 315, 46, 64, 82, 317, 100, 118, 136, 154, 319, 172, 190, 208, 226, 244, 262, 280, and 298; a nucleotide sequence encoding a polypeptide having the amino acid sequence of any of SEQ ID NOs: 27, 314, 45, 63, 81, 316, 99, 117, 135, 153, 318, 171, 189, 207, 225, 243, 261, 279, and 297; or a nucleotide sequence encoding a polypeptide having at least 90% identity to the amino acid sequence of any of SEQ ID NOs: 27, 314, 45, 63, 81, 316, 99, 117, 135, 153, 318, 171, 189, 207, 225, 243, 261, 279, and 297.

[0150] Also provided herein are vectors or constructs containing such nucleotide sequences. In some embodiments, the vectors or constructs contain one or more heterologous promoters operably linked to nucleotides encoding alpha or gamma chains and / or beta or delta chains. In some embodiments, the heterologous promoters are operably linked to one or more nucleotide sequences.

[0151] In some embodiments, a vector or construct can contain a single heterologous promoter driving the expression of one or more nucleotide sequences. In some embodiments, such a promoter can be multicistronic (e.g., bicistronic or tricistronic; see, e.g., U.S. Pat. No. 6,060,273). For example, in some embodiments, a transcription unit can be engineered as a bicistronic unit containing an IRES (internal ribosome entry site), allowing for the simultaneous expression of gene products (e.g., encoding the alpha or gamma and / or beta or delta chains of a TCR) by messages from a single promoter. Alternatively, in some cases, a single promoter can direct the expression of an RNA containing two or three genes (e.g., encoding the alpha or gamma and / or beta or delta chains of a TCR) in a single open reading frame (ORF), separated from each other by sequences encoding a self-cleaving peptide (e.g., a 2A peptide, e.g., a P2A peptide) or a protease recognition site (e.g., furin). An ORF can encode a single polyprotein that is cleaved into individual proteins either during translation (in the case of 2A, e.g., P2A) or post-translationally. In some cases, peptides such as P2A can cause the ribosome to skip synthesis of the peptide bond at the C-terminus of the 2A element (ribosomal skipping), resulting in separation between the end of the 2A sequence and the next downstream peptide (see, e.g., de Felipe. Genetic Vaccines and Ther. 2:13 (2004) and de Felipe et al. Traffic 5:616-626 (2004)). Examples of 2A cleavage peptides, including those that can induce ribosomal skipping, include Thosea asigna virus (T2A), porcine teschovirus-1 (P2A, e.g., SEQ ID NO: 309), equine rhinitis A virus (E2A), and the 2A sequence (F2A) from foot-and-mouth disease virus, as described in U.S. Patent Publication No. 2007 / 0116690.In some embodiments, the peptide that causes ribosome skipping is a P2A peptide and / or contains the sequence of amino acids set forth in SEQ ID NO:309.

[0152] In a bicistronic vector, the nucleic acid sequence encoding the alpha or gamma chain and the nucleotide sequence encoding the beta or delta chain can be present in any order and are separated by a nucleotide sequence encoding a self-cleaving peptide or peptide sequence that causes ribosome skipping (e.g., a P2A sequence). For example, in some embodiments, the nucleotide sequence comprises, in that order, a nucleic acid sequence encoding a beta or delta chain, a nucleic acid sequence encoding a self-cleaving peptide or peptide sequence that causes ribosome skipping (e.g., a P2A sequence described herein), and a nucleic acid sequence encoding an alpha or gamma chain. In other embodiments, the nucleotide sequence contains, in that order, a nucleic acid sequence encoding an alpha or gamma chain, a nucleic acid sequence encoding a self-cleaving peptide or peptide sequence that causes ribosome skipping (e.g., a P2A sequence described herein), and a nucleic acid sequence encoding a beta or delta chain.

[0153] In some embodiments, the nucleotide sequence encoding the TCR alpha or gamma chain and / or beta or delta chain comprises a nucleic acid sequence corresponding to a SEQ ID NO: set forth in Table 3. Also among the provided nucleotide sequences encoding TCRs are those containing sequences that are at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences. Also provided are any of the mature TCR alpha chains or mature TCR gamma chains encoded by any of the sequences set forth in Table 3, e.g., in each row therein. Also provided are any of the mature TCR beta chains or mature TCR delta chains encoded by any of the sequences set forth in Table 3, e.g., in each row therein. Also provided are any of the mature TCR alpha and beta chains, or mature gamma and delta chains encoded by any of the sequences set forth in Table 3, e.g., in each row therein. In some aspects, the nucleotide sequence contains a sequence encoding a signal sequence, and the encoded exemplary TCR, when expressed as a mature protein, includes, e.g., a mature Vα region and / or mature Vβ region, or a mature Vγ region and / or Vδ mature region that, when fully processed and expressed, does not include the signal sequence (e.g., from cleavage of the signal sequence).

[0154] In some embodiments, the nucleotide sequence encodes a polypeptide containing an amino acid sequence set forth in Table 3, such as in each row therein, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In some embodiments, the nucleotide sequence encodes a mature polypeptide set forth herein, e.g., in Table 3, such as in each row therein, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. SEQ ID NOs: 28, 46, 64, 82, 100, 118, 136, 154, 172, 190, 20, 226, 244, 262, 280, 298 encode, in order, the alpha chain, the P2A peptide, and the beta chain. In some embodiments, the nucleic acid sequences encode, in order, the beta chain, the P2A peptide, and the alpha chain (e.g., SEQ ID NO: 314, 316, or 318).

[0155] [Table 3] * Alternatively, the sequence can also be constructed as beta-P2A-alpha

[0156] Also provided herein are vectors, such as vectors containing any of the nucleic acids described herein. In some embodiments, a nucleic acid or nucleic acids encoding one or both chains of a TCR are cloned or assembled into a suitable expression vector or vectors. The expression vector may be any suitable recombinant expression vector and may be used to transform or transfect any suitable host. Suitable vectors include vectors designed for propagation and expansion, or for expression, or both, such as plasmids and viruses. In some embodiments, the vector is an expression vector.

[0157] III. Methods for Isolating, Characterizing, and Identifying T Cell Receptors In some aspects, provided herein are methods for isolating a plurality of nucleic acid sequences encoding provided TCRs specific for the minor histocompatibility antigen HA-2. In some aspects, the provided HA-2-specific TCRs are identified based on the methods described herein. In some aspects, the methods also include isolating the nucleic acid sequences, assembling the nucleic acid sequences into a vector, assessing the expression and / or activity of the TCRs, and optionally screening and identifying the particular TCRs of interest using high-throughput methods.

[0158] The methods described herein also relate to determining the binding activity and functional capacity of the candidate TCRs.

[0159] A. TCR Donor Criteria and Methods for Isolating miHA-Specific T Cell Candidates In some embodiments, whole exome sequencing is used to classify donors for their HLA repertoire and / or haplotype, as well as for polymorphisms encoding the miHA HA-2 antigen or non-immunogenic variants thereof. In some embodiments, the donor subject has cancer. In some embodiments, the donor subject has a type of cancer to be treated with engineered T cells. Subjects with the appropriate HLA type are selected for blood collection. Anti-HA-2 reactive T cells from the donor subject are identified based on their binding to HLA multimers folded with the antigenic HA-2 peptide. These cells are single-cell sorted, and their TCRs are screened for anti-HA-2 reactivity. In some embodiments, the donor has HA-2 (V peptide) + and HLA-A * 02:01 + is.

[0160] B. High-Throughput Isolation, Amplification, and Assembly of TCR-Encoding Nucleic Acid Sequences In some embodiments, nucleic acid molecules encoding TCRs can be obtained or identified from a variety of sources. In some embodiments, TCRs can be obtained or identified using high-throughput TCR isolation and screening methods. Examples of such methods that can be used include, for example, those described in WO 2018 / 102473, which is incorporated by reference in its entirety. In some embodiments, high-throughput TCR isolation and screening methods involve amplification of nucleic acids encoding TCR alpha and / or beta chains or TCR gamma and / or delta chains from a plurality of different cells, such as T cells isolated from a donor. Also provided herein are methods related to isolating or screening a plurality of different TCRs to obtain TCRs specific for relatively hematopoietic-restricted minor histocompatibility antigens, such as HA-2 peptides.

[0161] In some embodiments, nucleic acid molecules encoding TCRs can be obtained from a variety of sources, such as by polymerase chain reaction (PCR) amplification of encoding nucleic acids within or isolated from a given cell or cells. In some embodiments, TCRs are obtained from biological sources, such as cells such as T cells (e.g., cytotoxic T cells), T cell hybridomas, or other publicly available sources. In some embodiments, TCRs can be derived from one of a variety of animal species, such as human, mouse, rat, or other mammals. In some embodiments, T cells can be obtained from cells isolated in vivo, such as from a normal (or healthy) or diseased subject, including T cells present in peripheral blood mononuclear cells (PBMCs) or tumor-infiltrating lymphocytes (TILs). In some embodiments, T cells can be cultured T cell hybridomas or clones. For example, in some embodiments, to generate a vector encoding a TCR, the alpha and beta chains can be PCR amplified from total cDNA isolated from a T cell clone expressing the TCR of interest and cloned into an expression vector. In some embodiments, the alpha and beta chains can be produced synthetically. In some embodiments, the alpha and beta chains are cloned into the same vector.

[0162] As described herein, the methods and materials provided herein may enable users to successfully capture most, if not all, functional TCRs from a sorted T cell population. For example, an amplification (e.g., a nested amplification procedure such as nested PCR) procedure may involve using a primer collection designed to amplify all known functional V segments of the two variable chains of a particular mammalian (e.g., human) TCR (e.g., any known functional V segments of the α and β variable chains of a particular αβ TCR, or any known functional V segments of the γ and δ variable chains of a particular γδ TCR). For humans, the amplification procedure may include a primer collection designed to amplify all 45 V segments of the α chain currently known to be functional and all 48 V segments of the β chain currently known to be functional. When referring to a TCR V segment of the α chain herein, the abbreviation TRAV may be used. Similarly, when referring to a TCR V segment of the β chain herein, the abbreviation TRBV may be used. The same is true for the TCR V-segments of the gamma and delta chains, which can be referred to as TRGV and TRDV, respectively.

[0163] In some aspects, this document provides methods and materials related to the cloning of functional TCRs from single T cells. For example, in some aspects, this document provides methods and materials for obtaining nucleic acid encoding a TCR from a single T cell and arranging the nucleic acid to form a nucleic acid vector successfully designed to express a TCR (e.g., a fully intact TCR, such as a fully intact TCR with the variable chain combination present in the single T cell), kits for obtaining nucleic acid encoding a TCR from a single T cell and arranging the nucleic acid to form a nucleic acid vector successfully designed to express a TCR (e.g., a fully intact TCR, such as a fully intact TCR with the variable chain combination present in the single T cell), and methods for making such kits. A cloned αβ TCR with a variable chain combination as present in the single T cell used to clone the TCR can include a VJα segment combination as present in the single T cell, a VDJβ segment combination as present in the single T cell, the nucleotide sequence of the entire α variable region as present in the single T cell, and the nucleotide sequence of the entire β variable region as present in the single T cell. Similarly, a cloned γδ TCR having a variable chain combination as present in the single T cell used to clone the TCR may comprise a VJγ segment combination as present in that single T cell, a VDJδ segment combination as present in that single T cell, the nucleotide sequence of the entire gamma variable region as present in that single T cell, and the nucleotide sequence of the entire delta variable region as present in that single T cell.

[0164] In some aspects, this document provides collections of nucleic acid primers designed to amplify the entire coding sequence of both variable regions (e.g., the α and β variable regions, or the γ and δ variable regions) of each expressed V-segment (e.g., each expressed α and β V-segment, or each expressed γ and δ V-segment) of a functional αβ or γδ TCR of a particular mammalian species (e.g., mouse or human), methods of using such collections of nucleic acid primers to clone functional TCRs from single T cells, and kits containing such collections of nucleic acid primers for cloning functional TCRs from single T cells.

[0165] In some aspects, the methods and materials provided herein may allow for highly multiplexed reactions to be performed, allowing for the cloning of many different TCRs (e.g., hundreds to thousands, or more, of different TCRs) directly from a single T cell rapidly (e.g., in some cases simultaneously) and in a manner that results in little, if any, loss of α / β variable chain combinations (or γ / δ variable chain combinations). For example, the methods and materials provided herein can be used to clone many different αβ TCRs (e.g., hundreds to thousands or more different αβ TCRs) directly from a single αβ T-cell in a manner that loses less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent) of the α variable chains and less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent) of the β variable chains possible for a species (e.g., a mouse or human species). Similarly, the methods and materials provided herein can be used to clone many different γδ TCRs (e.g., hundreds to thousands or more different γδ TCRs) directly from a single γδ T cell in a manner that loses less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent) of the gamma variable chains and less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent) of the delta variable chains possible for a species (e.g., a mouse or human species). In some cases, the methods and materials provided herein involve (a) obtaining a sample of T cells; (b) sorting those T cells into isolated locations (e.g., wells) so that most, if not all, of the isolated locations (e.g., each well) contain a single T cell; and(c) lysing (e.g., simultaneously lysing) single T cells located in separate isolated locations (e.g., separate wells) to release the RNA of each single T cell; (d) performing (e.g., simultaneously) reverse transcription using the released RNA as a template, appropriate primers for cDNA synthesis from the RNA, and reverse transcriptase to produce cDNA within each isolated location (e.g., each well), wherein the cDNA represents the RNA expressed by the single T cell located in that isolated location (e.g., well); and (e) performing reverse transcription on each isolated T cell using the released RNA as a template, appropriate primers for cDNA synthesis from the RNA, and reverse transcriptase to produce cDNA within each isolated location (e.g., each well), wherein the cDNA represents the RNA expressed by the single T cell located in that isolated location (e.g., well). For the isolated locations, a first round amplification reaction (e.g., a first round polymerase chain reaction (PCR)) is performed (e.g., simultaneously) using the produced cDNA as a template, a first round primer collection (e.g., a first round PCR primer collection), and a polymerase (e.g., Taq polymerase) in an amplification procedure (e.g., a nested amplification procedure such as nested PCR) to generate an amplification product containing the nucleic acid sequence of the α variable chain (or γ variable chain) of the TCR of a single T cell at the isolated location, and a nucleic acid sequence of the same single T cell. (f) for each isolated location, performing (e.g., simultaneously performing) a second round amplification reaction (e.g., second round PCR) of a nested amplification procedure (e.g., nested PCR procedure) using the amplification product of the first round amplification reaction as a template, a second round primer collection (e.g., a second round PCR primer collection), and a polymerase (e.g., Taq polymerase). ), producing at least a first amplification product containing the nucleic acid sequence of the α variable chain (or γ variable chain) of the TCR of the single T cell at that isolated location, and a second amplification product containing the nucleic acid sequence of the β variable chain (or δ variable chain) of the TCR of that same single T cell at that same isolated location; and (g) for each isolated location, combining the first amplification product and the second amplification product with an α / β variable chain or a γ / δ variable chain combination (or V segment of an α / β variable chain or a γ / δ variable chain combination) as was present in the single T cell used to generate the amplification product.and cloning the TCR into an expression vector designed to express a functional TCR having the TCR fragment (part thereof).

[0166] The resulting expression vector can be introduced into cells so that the cells express the cloned TCR. Such cells, and / or the TCRs they express from the introduced expression vector, can be screened to identify TCRs with desired capabilities. For example, cells expressing a cloned TCR that recognizes a particular antigen (e.g., a peptide derived from a tumor polypeptide) can be identified, and those cells, the TCR expression vectors they contain, or the cloned TCR constructs can be used for further analysis or therapeutic applications.

[0167] In some cases, once the signaling machinery of a functional TCR is activated, expression of the cloned TCR on its surface and expression of a functional TCR can be assessed by introducing the expression vector into TCR-negative reporter cells engineered to express a measurable marker signal or marker polypeptide. In these cases, functional TCRs can be screened using antibodies (e.g., anti-CD3 antibodies) designed to nonspecifically activate the TCR. In some cases, cloned TCRs can be screened for antigen specificity. For example, reporter cells expressing the cloned TCR can be screened for recognition of a specific antigen (e.g., a peptide derived from a tumor polypeptide). In some cases, primary T cells (e.g., human primary T cells) can be transfected with the expression vector and screened for antigen specificity via a T cell proliferation assay.

[0168] The methods and materials provided herein may enable clinicians, medical professionals, laboratory personnel, and researchers to use a collection of T cells with different TCRs to generate a collection of expression vectors that express functional versions of different TCRs with the same variable chain combinations or portions thereof (e.g., the same α / β variable chain combinations or the same γ / δ variable chain combinations) that were present in the original T cells used to generate the collection. Such a collection of expression vectors can be obtained quickly, efficiently, inexpensively, and effectively. For example, in some cases, using the methods and materials provided herein, a collection of expression vectors expressing functional versions of many different TCRs with authentic variable chain combinations as found in T cells obtained from a mammal (e.g., a human) can be generated in less than 12 days (e.g., 4-11 days, 5-11 days, 6-11 days, 7-11 days, 8-11 days, 4-10 days, 5-10 days, 6-10 days, 7-10 days, 8-10 days, 4-9 days, 5-9 days, 6-9 days, 7-9 days, 4-8 ...5-9 days, 6-9 days, 7-9 days, 4-8 days, 5-9 days, 5-9 days, 6-9 days, 7-9 days, 4-8 days, 5-9 days, 5-9 days, 6-9 days, 7-9 days, 4-8 days, 5-9 days, 5-9 days, 6-9 days, 7-9 days, 5-9 days, 5-9 days, 6-9 days, 7-9 days, 5-9 days, 5-9 days, 6-9 days, 7-9 days, 5-9 days, 5-9 TCRs can be generated within 8, 6, or 8 days, using fewer than 12 steps (e.g., 5-11 steps, 6-11 steps, 7-11 steps, 8-11 steps, 5-10 steps, 6-10 steps, 7-10 steps, 8-10 steps, 5-9 steps, 6-9 steps, 7-9 steps, or 8-9 steps), for less than about $10 per TCR, and with greater than about 80 percent (e.g., greater than about 85, 90, or 95 percent) efficacy (based on sorting a single T cell into each of 384 wells of a 384-well plate). In some cases, the methods and materials provided herein can be performed without nucleic acid sequencing, without a restriction endonuclease cleavage step, without other steps or techniques described herein, and / or without the use of certain reagents or materials described herein.

[0169] The methods and materials provided herein may also enable users to successfully capture most, if not all, functional TCRs from a sorted T cell population. For example, in some cases, the methods and materials provided herein may include a nested amplification procedure (e.g., a nested PCR procedure) that includes a primer collection designed to amplify all known functional V segments of the two variable chains of a particular mammalian (e.g., human) TCR (e.g., any of the known functional V segments of the α and β variable chains of a particular αβ TCR, or any of the known functional V segments of the γ and δ variable chains of a particular γδ TCR). Having the ability to clone most, if not all, functional TCRs from a sorted T cell population may enable users to identify specific TCRs, including rare TCRs, that might otherwise be overlooked. It is these rare TCRs that may be overlooked that can provide a rich source of newly cloned TCRs for effective therapies, such as cancer treatments, involving the delivery of effective T cells.

[0170] In some cases, the methods and materials provided herein may enable users to obtain additional information about single T cells from which functional TCR clones are generated. In some cases, the flow cytometry techniques used for single-cell sorting described herein can be used to distinguish activated and experienced T cells from naive T cells by staining the cells for activation markers. When applying the methods and materials provided herein in methods for treating certain diseases (e.g., cancer), T cells can be isolated from a patient in which they have already been activated and expanded. Once these T cells are isolated and cDNA is generated from single-cell RNA, an additional level of selection can be applied. For example, in addition to using cDNA generated from the RNA of a single T cell to amplify and clone the variable chain (or portion thereof) of the TCR of that T cell, the cDNA can also be used to assess RNA expression and / or RNA expression levels within that T cell.

[0171] CD8 + In the case of T cells, TCRs associated with multifunctional (e.g., multi-cytokine producing) effector cells or TCRs associated with resting or exhausted long-lived memory cells can be identified by examining the relative mRNA levels of expression of transcription factors such as Eomesodermin and T-bet (McLane et al., J. Immunol., 190(7):3207-3215 (2013); and Buggert et al., PLoS Pathog., 10(7):e1004251 (2014)).

[0172] In some cases, T cells can be stimulated (e.g., in vitro) before sorting, and RNA expression can then be assessed (e.g., via qPCR) to determine which T cells responded to the stimulation. Any suitable type of stimulation can be used, including, but not limited to, non-specific stimulation, such as with concanavalin A, phytohemagglutinin-P, phorbol ester plus ionomycin, phorbol myristate acetate plus calcium ionophore, or an antibody capable of cross-linking the TCR (e.g., an anti-CD3 antibody plus an anti-CD28 antibody, or an anti-TCRβ antibody), or antigen-specific stimulation, such as with one or more specific antigens as described elsewhere (Downward et al., Nature, 346:719-23 (1990); and Dasgupta et al., Proc. Natl. Acad. Sci. USA, 84:1094-8 (1987)). In some cases, cytokine expression levels, such as TNF-α, IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-13, or IL-17 expression levels, can be determined and compared with unstimulated populations. Once single T cells are sorted, the methods provided herein can be used to determine which T cells were making specific cytokines in response to stimulation (e.g., in response to a peptide antigen used to stimulate the T cells). In these cases, antigen-specific T cells can be determined without the tedious method of expanding reactive T cells or the destructive method of paraformaldehyde fixation and intracellular cytokine staining, which can reduce the ability to effectively clone TCRs. In such cases, specific TCRs generated from activated and antigen-specific T cells can be rapidly identified, as opposed to inactive bystander T cells.

[0173] In some cases, cytokine expression levels, such as TNF-α, IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-13, or IL-17 expression levels, can be determined for a single T cell used to clone a functional TCR, thereby allowing for identification of a particular TCR based on the particular phenotype (e.g., elevated IFN-γ expression) of the T cell that donated the variable chain (or portion thereof) of the particular TCR. In such cases, specific TCRs generated from active, rather than inactive, T cells can be rapidly identified. In some cases, specific TCRs generated from inactive, rather than active, T cells can be rapidly identified.

[0174] In some cases, the absence of cytokine production by a T cell does not necessarily reflect the absence of TCR specificity. Signals initiated by the TCR on a cell can be disrupted and / or suppressed by a number of inhibitory co-receptors (Sheppard et al., FEBS Lett., 574(1-3):37-41 (2004); and Yokosuka et al., J. Exp. Med., 209(6):1201-1217 (2012)). In some cases, TCRs can be obtained using T cells that are refractory to stimulation, and the specificity of the cloned TCR can be tested or screened in cells in which canonical TCR signaling is not suppressed.

[0175] In some cases, MHC-peptide complexes (or HLA-peptide complexes) can be used to identify cloned TCRs that recognize such complexes. In these cases, it is possible that clonal exclusion and / or lack of antigen priming during the immune response can result in TCRs with this specificity not being present in the activated and / or expanded TCR pool. In such cases, the methods and materials provided herein can be used to clone naive or inactivated TCRs that recognize such complexes, although in some cases only a single T cell needs to be present. In some cases, a pool of naive T cells can be stained with MHC-peptide tetramers (or HLA-peptide tetramers), and any MHC-peptide (or HLA-peptide)-reactive TCRs in the naive T cells can be used to clone these TCRs using the methods and materials provided herein.

[0176] In some aspects, methods provided herein include methods for obtaining a plurality of nucleic acid vectors containing nucleic acids encoding functional T cell receptors, the method comprising, or consisting essentially of: (a) obtaining a device including a plurality of distinct locations, each distinct location containing cDNA generated from RNA obtained from a single T cell sorted to the distinct location; (b) performing a nested amplification procedure using the cDNA at each of the plurality of distinct locations as a template to obtain a first amplification product and a second amplification product for each cDNA at the plurality of distinct locations, wherein the first amplification product comprises a nucleic acid encoding a Vα segment or a Vγ segment and the second amplification product comprises a nucleic acid encoding a Vβ segment or a Vδ segment; and (c) assembling the first amplification product and the second amplification product for each cDNA at the plurality of distinct locations into a nucleic acid vector to obtain an assembled nucleic acid vector for each cDNA at the plurality of distinct locations, wherein the assembled nucleic acid vector for each cDNA at the plurality of distinct locations comprises a nucleic acid encoding a functional T cell receptor. The plurality can be greater than 50. The plurality can be greater than 500. The plurality can be greater than 5000. The plurality of nucleic acid vectors can be a plurality of nucleic acid expression vectors. The device can comprise a multiwell plate. The multiwell plate can be a 96-well plate, a 384-well plate, or a 1536-well plate. The cDNA generated from RNA obtained from a single T cell can comprise cDNA generated from RNA obtained from a single human T cell. The first amplification product can comprise a nucleic acid encoding the L sequence of a Vα or Vγ segment. The first amplification product can comprise a nucleic acid encoding a Jα or Jγ segment. The first amplification product can comprise a nucleic acid encoding a 5' portion of a Cα or Cγ region. The first amplification product can comprise a nucleic acid encoding the L sequence of a Vα or Vγ segment, a Jα or Jγ segment, and a 5' portion of a Cα or Cγ region. The second amplification product can comprise a nucleic acid encoding the L sequence of a Vβ or Vδ segment.The second amplification product may include a nucleic acid encoding a Dβ segment or a Dδ segment. The second amplification product may include a nucleic acid encoding a Jβ segment or a Jδ segment. The second amplification product may include a nucleic acid encoding a 5' portion of a Cβ or Cδ region. The second amplification product may include a nucleic acid encoding a Vβ or Vδ segment, a Dβ or Dδ segment, a Jβ or Jδ segment, and an L sequence in the 5' portion of a Cβ or Cδ region. The first amplification product may include an adapter sequence added to the cDNA amplification template sequence via the second round of amplification in the nested amplification procedure. The second amplification product may include an adapter sequence added to the cDNA amplification template sequence via the second round of amplification in the nested amplification procedure. The first amplification product may include a first adapter sequence added to the cDNA amplification template sequence via a second round of amplification in a nested amplification procedure, and the second amplification product may include a second adapter sequence added to the cDNA amplification template sequence via a second round of amplification in a nested amplification procedure, wherein the first adapter sequence and the second adapter sequence are different. Each functional T cell receptor of the assembled nucleic acid vector may include a Vα / Vβ combination or a Vγ / Vδ combination as present in a single T cell originating from the RNA. Each functional T cell receptor of the assembled nucleic acid vector may include (a) a full-length α variable region and a full-length β variable region, or (b) a full-length γ variable region and a full-length δ variable region. Each functional T cell receptor of the assembled nucleic acid vector may include (a) a full-length α variable region and a full-length β variable region as present in a single T cell originating from the RNA, or (b) a full-length γ variable region and a full-length δ variable region as present in a single T cell originating from the RNA. The functional T cell receptor of each assembled nucleic acid vector can include (a) a full-length alpha constant region and a full-length beta constant region, or (b) a full-length gamma constant region and a full-length delta constant region. Each assembled nucleic acid vector can include a nucleic acid sequence encoding a self-cleaving peptide or an internal ribosome entry site (IRES). The method can include sorting single T cells into distinct locations.The method may include performing a reverse transcription reaction to obtain cDNA. The assembling step may include seamless cloning. Each assembled nucleic acid vector may be obtained without performing nucleic acid sequencing. Each assembled nucleic acid vector may be obtained without performing a restriction endonuclease cleavage reaction.

[0177] C. Assessment of Minor Histocompatibility Antigen-Specific T Cell Receptor Expression, Activity, and Function Exemplary assays can be used to assess the activity, expression, and / or function of the TCRs and antigen-binding fragments described herein. The assays described herein should not be construed as limiting and can be used to assess the functional capabilities of candidate miHA-specific TCRs.

[0178] Functional characterization of TCRs can be performed either by binding assays utilizing fluorescently labeled MHC molecules bearing specific target peptides (tetramers / pentamers / dextramers) or by activation assays by co-culturing TCR-expressing cells with antigen-presenting cells (APCs) presenting the corresponding MHC / peptide complexes.

[0179] Cytokine release assays can assess the ability of candidate TCRs to produce the cytokines IL-2 and / or IFN-γ after exposure to cells presenting the target antigen. T cells were incubated with T2 cells loaded with the target HA-2 "V" peptide (HA-2 (neg) / HLA-A * :02:01 (pos) ) as controls. As controls, T2 cells are loaded with the non-targeting HA-2 "M" peptide or an irrelevant peptide control. T cell IL-2 and / or IFN-γ responses are followed by intracellular cytokine staining and analysis by FACS.

[0180] A T cell activation / degranulation marker assay can be used to assess the ability of candidate TCRs to express the surface marker CD107a after exposure to cells presenting a target antigen. CD107a is a marker for T cell degranulation, which is part of the cell killing response. T cells are incubated with T2 cells loaded with the target HA-2 "V" peptide or a control peptide, such as the non-target HA-2 "M" peptide or an irrelevant peptide. The degranulation response is tracked on the T cells by CD107a surface staining and analysis by FACS.

[0181] Killing assays can evaluate the ability of candidate TCRs to lyse cells presenting target antigens. T cells are incubated with a mixture of fluorescently tagged T2 cells differentially loaded with target and control peptides, allowing on-target and off-target cytotoxicity to be tested within a single test sample. Fluorescent cell counting beads are included as a normalization / counting control. Fluorescently tagged T2 cells are loaded with the target HA-2 "V" peptide. As a control, T2 cells labeled with a different fluorescent tag are loaded with the non-target HA-2 "M" peptide or an irrelevant peptide control. Gated cell counts of HA-2 "V" peptide-loaded T2 cells and control peptide-loaded T2 cells remaining after incubation with T cells are tracked by FACS.

[0182] The CD34 marker can be used as a surrogate potency measure. See Philip et al. (2014) Blood 124(8):1277-1287. Detection of CD34, a marker of transduction efficiency, can be correlated with the functional potency measures described herein.

[0183] In some embodiments, the expansion screening and non-expansion screening are performed using the exact same donors for direct comparison of the methods. Expansion can be performed using a specific method. Methods that generate candidate TCRs without an expansion process can be advantageous in some situations, given the time sensitivity of screening for TCRs with the desired specificity. Reduced sample processing can also provide advantages in different situations.

[0184] IV. Engineered Cells Also provided herein are cells, such as cells engineered to contain a TCR as described herein. Also provided herein are populations of such cells, as well as compositions containing and / or enriched for such cells, where the TCR-expressing cells constitute at least 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more percent of the total cells in the composition. In some embodiments, the cells are primary T cells, or T cells or CD8 + Cells or CD4 + The compositions are specific types of cells, such as cells. Among the compositions are pharmaceutical compositions and formulations for administration, such as adoptive cell therapy. Therapeutic methods for administering the cells and compositions to a subject, e.g., a patient, are also provided herein.

[0185] Thus, also provided herein are genetically engineered cells that express the TCRs provided herein. The cells are generally eukaryotic cells, such as mammalian cells, and typically human cells. In some embodiments, the cells are derived from blood, bone marrow, lymph, or lymphoid organs and are cells of the immune system, such as cells of the innate or adaptive immune system, e.g., myeloid cells or lymphoid cells, including lymphocytes, typically T cells and / or NK cells. Other exemplary cells include stem cells, such as multipotent and pluripotent stem cells, including induced pluripotent stem cells (iPSCs). The cells are typically primary cells (e.g., primary T cells), such as those isolated directly from a subject (e.g., a donor subject) and / or isolated and frozen from a subject. In some embodiments, the cells include one or more subsets of T cells or other cell types, such as the total T cell population, CD4+ cells, CD8+ cells, and subpopulations thereof, defined by, for example, function, activation state, maturity, differentiation potential, expansion, recirculation, localization, and / or persistence, antigen specificity, antigen receptor type, presence in a particular organ or compartment, marker or cytokine secretion profile, and / or degree of differentiation. With respect to the subject being treated, the cells can be allogeneic and / or autologous. The methods provided herein include off-the-shelf methods. In some aspects, such as for off-the-shelf techniques, the cells are pluripotent and / or multipotent, such as stem cells (e.g., iPSCs). In some embodiments, the methods provided herein include isolating cells from a subject, preparing, treating, culturing, and / or manipulating them as described herein, and reintroducing them into the same patient, before or after cryopreservation.

[0186] T cells (including primary T cells) and / or CD4 + T cells and / or CD8 + T cells (primary CD4 + T cells and / or CD8 + Among the subtypes and subpopulations of T cells, including naive T (TN ) cells, effector T cells (T EFF ), memory T cells and their subtypes, such as stem cell memory T (T SCM ), Central Memory T(T CM ), Effector Memory T(T EM ), or terminally differentiated effector memory T cells, tumor-infiltrating lymphocytes (TILs), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosa-associated invariant T (MAIT) cells, natural and adaptive regulatory T (Treg) cells, helper T cells, such as TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, alpha / beta T cells, and delta / gamma T cells.

[0187] In some embodiments, the cell is a NK cell, hi some embodiments, the cell is a monocyte or granulocyte, e.g., a myeloid cell, a macrophage, a neutrophil, a dendritic cell, a mast cell, an eosinophil, and / or a basophil.

[0188] In some embodiments, the cells contain one or more nucleic acids introduced via genetic engineering, thereby expressing recombinant or engineered products of such nucleic acids. In some embodiments, the nucleic acids are heterologous, i.e., not normally present in the cell or sample obtained from the cell, e.g., obtained from another organism or cell not normally found in the cell being engineered and / or the organism from which such cell is derived. In some embodiments, the nucleic acids are non-naturally occurring, such as nucleic acids not found in nature, including those containing chimeric combinations of nucleic acids encoding various domains from multiple different cell types.

[0189] In some embodiments, expression of endogenous TCR chains of engineered cells is reduced or eliminated, e.g., to reduce the risk or likelihood of mispairing between the chains of the engineered TCR and the endogenous TCR. Such mispairing may create new TCRs that could potentially result in a higher risk of undesired or unintended antigen recognition and / or side effects, and / or may reduce the expression level of the desired exogenous TCR. Exemplary methods for reducing or preventing endogenous TCR expression are described elsewhere, see, e.g., U.S. Pat. No. 9,273,283; U.S. Patent Application Publication No. 2014 / 0301990.

[0190] In some embodiments, a nucleic acid encoding an anti-HA-2 CAR is transfected into T cells to form anti-HA-2 CAR T cells. The T cells can be, but are not limited to, primary T cells, cultured T cells, autologous T cells, allogeneic T cells, T cells obtained from bone marrow, T cells obtained from lymph nodes, T cells obtained from the thymus, tumor-infiltrating lymphocytes, T cells obtained from the spleen, T cells from umbilical cord blood, universal allogeneic T cells, universal CAR T cells, CAR T cells, naive T cells, effector T cells, effector memory T cells, CD4+ / CD8+ T cells, helper T cells, CD4+ T cells, CD4+ helper T cells, Th1 T cells, Th2 T cells, cytotoxic T cells, CD8+ T cells, peripheral blood mononuclear cells (PBMCs), peripheral blood leukocytes (PBLs), memory T cells, central memory T cells, regulatory T cells, αβ T cells, γδ T cells, modified T cells, T cells for use in adoptive cell transfer therapy, or TCR-engineered T cells. The CAR T cells can be first generation CAR T cells, second generation CAR T cells, third generation CAR T cells, fourth generation CAR T cells, dual antigen receptor CAR T cells, or CAR T cells with an inducible suicide gene, or a combination thereof.

[0191] A. Preparation of Cells for Genetic Manipulation In some embodiments, preparation of engineered cells involves one or more culture and / or preparation steps. Cells for introduction of a TCR can be isolated from a sample, such as a biological sample, e.g., a sample obtained or derived from a subject. In some embodiments, the subject from which the cells are isolated is a subject with a disease or condition, or a subject in need of cell therapy, or a subject to whom cell therapy will be administered. In some embodiments, the subject is a human in need of a particular therapeutic intervention, such as adoptive cell therapy, from which cells have been isolated, processed, and / or engineered.

[0192] In some embodiments, the engineered cells are derived from a donor subject (i.e., the donor subject is not the recipient subject) who is not a subject with a disease or condition, or a subject in need of cell therapy, or a subject to whom cell therapy will be administered. In some embodiments, the donor subject is HLA-matched to the recipient subject. In some embodiments, the donor subject does not express the miHA-2 "M" antigen.

[0193] Thus, in some embodiments, the cells are primary cells, e.g., primary human cells. Samples include tissues, body fluids, and other samples taken directly from a subject, as well as samples obtained after one or more processing steps, such as separation, centrifugation, genetic manipulation (e.g., transduction with a viral vector), washing, and / or incubation. Biological samples can be samples obtained directly from a biological source or processed samples. Biological samples include, but are not limited to, body fluids, such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine, and sweat, tissue samples, and organ samples (including processed samples derived therefrom).

[0194] In some embodiments, the sample from which cells are derived or isolated is a blood or blood-derived sample, or an apheresis or leukapheresis product, or is derived therefrom. Exemplary samples include whole blood, PBMCs, white blood cells, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut-associated lymphoma tissue, mucosa-associated lymphoma tissue, pancreas, other lymphoma tissue, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testis, ovary, tonsil, or other organ, and / or cells derived therefrom. Samples include samples from autologous and allogeneic sources in the context of cell therapy, e.g., adoptive cell therapy.

[0195] In some embodiments, the cells are derived from a cell line, e.g., a T cell line. In some embodiments, the cells are obtained from a heterologous source, e.g., from a mouse, rat, non-human primate, or pig.

[0196] B. Vectors and Methods for Genetic Engineering Also provided herein are methods, nucleic acids, compositions, and kits for expressing the TCRs or antigen-binding fragments thereof provided herein in cells (e.g., genetically engineered cells), and for producing genetically engineered cells that express such TCRs or antigen-binding fragments thereof. Genetic engineering generally involves the introduction of a nucleic acid encoding the TCR (or antigen-binding fragment thereof) into a cell, such as by retroviral transduction, transfection, or transformation.

[0197] In some embodiments, gene transfer is achieved by first stimulating the cells, such as by combining them with a stimulus that induces a response such as proliferation, survival, and / or activation (e.g., as measured by expression of cytokines or activation markers), and then transducing the activated cells and expanding them in culture to numbers sufficient for clinical use.

[0198] Various methods for the introduction of genetically engineered components are well known and can be used with the provided methods and compositions. Exemplary methods include methods for the transfer of nucleic acids encoding the TCRs or antigen-binding fragments thereof provided herein, including via viral vectors (e.g., retroviruses or lentiviruses), transduction, transposons, and electroporation.

[0199] In some embodiments, the recombinant nucleic acid is transferred into the cell using a recombinant infectious viral particle. In some embodiments, the recombinant nucleic acid is transferred into the T cell using a recombinant lentiviral vector or a retroviral vector, such as a gammaretroviral vector (see, e.g., Koste et al. (2014) Gene Therapy 2014 Apr 3; Carlens et al. (2000) Exp Hematol 28(10):1137-46; Alonso-Camino et al. (2013) Mol Ther Nucl Acids 2,e93; Park et al., Trends Biotechnol. 2011 November 29(11):550-557).

[0200] In some embodiments, the retroviral vector has long terminal repeats (LTRs), such as those derived from Moloney murine leukemia virus (MoMLV), myeloproliferative sarcoma virus (MPSV), murine embryonic stem cell virus (MESV), murine stem cell virus (MSCV), or spleen focus forming virus (SFFV). In some embodiments, the retrovirus includes those derived from any avian or mammalian cell source. Retroviruses are typically amphotropic, meaning they can infect host cells of several species, including humans. In some embodiments, the nucleic acid to be expressed replaces the retroviral gag, pol, and / or env sequences. Several exemplary retroviral systems are described elsewhere (see, e.g., U.S. Patent Nos. 5,219,740; 6,207,453; 5,219,740; Miller and Rosman (1989) BioTechniques 7:980-990; Miller, AD (1990) Human Gene Therapy 1:5-14; Scarpa et al. (1991) Virology 180:849-852; Burns et al. (1993) Proc. Natl. Acad. Sci. USA 90:8033-8037; and Boris-Lawrie and Temin (1993) Cur. Opin. Genet. Develop. 3:102-109).

[0201] Methods for lentiviral transduction are known. Exemplary methods are described, for example, in Wang et al. (2012) J. 35(9):689-701; Cooper et al. (2003) Blood. 101:1637-1644; Verhoeyen et al. (2009) Methods Mol Biol. 506:97-114; and Cavalieri et al. (2003) Blood. 102(2):497-505.

[0202] In some embodiments, recombinant nucleic acids are transferred into T cells via electroporation (see, e.g., Chicaybam et al. (2013) PLoS ONE 8(3):e60298 and Van Tedeloo et al. (2000) Gene Therapy 7(16):1431-1437). In some embodiments, recombinant nucleic acids are transferred into T cells via transposition (see, e.g., Manuri et al. (2010) Hum Gene Ther 21(4):427-437; Sharma et al. (2013) Molec Ther Nucl Acids 2,e74; and Huang et al. (2009) Methods Mol Biol 506:115-126). Other methods for introducing and expressing the nucleic acids provided herein into immune cells include calcium phosphate transfection (e.g., as described in Current Protocols Molecular Biology, John Wiley & Sons, New York, NY), protoplast fusion, cationic liposome-mediated transfection, tungsten particle-facilitated microparticle bombardment (Johnston, Nature, 346:776-777 (1990)), and strontium phosphate DNA co-precipitation (Brash et al., Mol. Cell Biol., 7:2031-2034 (1987)).

[0203] Other approaches and vectors for the transfer of nucleic acids encoding the TCRs, antigen-binding fragments thereof, or recombinant products provided herein include those described elsewhere, see, e.g., International Patent Application Publication No. WO 2014 / 055668 and U.S. Patent No. 7,446,190.

[0204] In some cases, one or more additional nucleic acids can be introduced into cells simultaneously or sequentially with the nucleic acids encoding the TCRs or antigen-binding fragments thereof provided herein. In some cases, such additional nucleic acids for introduction can improve the efficacy of the treatment, e.g., by promoting the viability and / or function of the transferred cells; provide genetic markers for selection and / or evaluation of cells, e.g., to assess survival or localization in vivo; and / or improve safety, e.g., by making cells susceptible to negative selection in vivo, as described elsewhere (Lupton SD et al., Mol. and Cell Biol., 11:6 (1991); and Riddell et al., Human Gene Therapy 3:319-338 (1992)). See also (a) International Patent Applications PCT / US91 / 08442 and PCT / US94 / 05601 by Lupton et al., which describe the use of bifunctional selectable fusion genes by fusing a dominant positive selectable marker with a negative selectable marker, and (b) columns 14-17 of U.S. Pat. No. 6,040,177 to Riddell et al.

[0205] Thus, in some embodiments, engineered cells are provided, such as those containing a TCR or antigen-binding fragment thereof, nucleic acid, or vector described herein. In some aspects, the cells are generated by transducing a cell in vitro or ex vivo with a vector described herein. In some aspects, the cell is a T cell, such as a CD8+ or CD4+ T cell. In some embodiments, the TCR is heterologous to the cell.

[0206] V. Therapeutic and Prophylactic Methods and Uses Also provided herein are methods of administering the TCRs and antigen-binding fragments thereof, and / or engineered cells expressing the TCRs or antigen-binding fragments thereof, as provided herein, and uses, including therapeutic and prophylactic uses. Such methods and uses include, for example, therapeutic methods and uses involving administration of the molecules, cells, or compositions containing same to a subject having a miHA HA-2 YIGEVLVSV allele. In some embodiments, such methods and uses include, for example, therapeutic methods and uses involving administration of the molecules, cells, or compositions containing same to a subject having a hematological disease, condition, or disorder for which alloSCT is a treatment option. In some embodiments, alloSCT involves transplantation of hematopoietic cells that are insensitive to engineered T cells expressing any of the described anti-HA-2 TCRs. In some embodiments, such methods and uses include, for example, therapeutic methods and uses involving administration of the molecules, cells, or compositions containing same to a subject having a hematological disease, condition, or disorder for which alloSCT is not a treatment option. In some embodiments, the molecules, cells, and / or compositions are administered in an amount effective to provide treatment for a disease or disorder. Uses include the use of the TCRs and cells in such methods and treatments, and in the preparation of medicaments for carrying out such treatment methods. In some embodiments, the methods are carried out by administering the TCRs or cells, or compositions comprising same, to a subject who has, has had, or is suspected of having a disease or condition (e.g., hematopoietic cancer). In some embodiments, the methods thereby treat the disease or condition or disorder in the subject.

[0207] The described TCRs and engineered T cells expressing the described TCRs can be used to kill or eliminate autologous bone marrow-derived cells in subjects with miHA HA-2 YIGEVLVSV alleles. Killing or eliminating autologous bone marrow-derived cells in subjects with miHA HA-2 YIGEVLVSV alleles can be used to enhance alloSCT, treat hematologic malignancies, or treat non-hematologic diseases or conditions (e.g., autoimmune disorders or hematologic diseases). In some embodiments, alloSCT involves transplantation of hematopoietic cells that are insensitive to engineered T cells expressing any of the described anti-HA-2 TCRs.

[0208] As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") refers to the complete or partial amelioration or reduction of a disease or condition or disorder, or its associated symptoms, adverse effects, or outcomes, or phenotype. Desirable effects of treatment include, but are not limited to, prevention of disease onset or recurrence, alleviation of symptoms, reduction of any direct or indirect pathological consequences of disease, prevention of metastasis, reduction in the rate of disease progression, improvement or palliation of the disease state, and remission or improved prognosis. These terms do not imply complete cure of a disease or complete elimination of any symptoms or effects on all symptoms or outcomes.

[0209] As used herein, "delaying the onset of disease" means postponing, preventing, slowing, delaying, stabilizing, inhibiting, and / or postponing the onset of a disease (such as cancer). This delay can be of varying lengths of time, depending on the history of the disease and / or the individual being treated. As will be apparent to one of skill in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease. For example, late-stage cancer, such as the development of metastases, can be delayed.

[0210] As used herein, "preventing" includes providing protection against the occurrence or recurrence of a disease in a subject who may be susceptible to the disease but has not yet been diagnosed with the disease. In some embodiments, the provided molecules and compositions are used to delay the onset of the disease or slow the progression of the disease.

[0211] As used herein, "inhibiting" a function or activity refers to decreasing the function or activity when compared to conditions that are otherwise the same except for the condition or parameter of interest, or when compared to another condition. For example, a TCR or composition or cell that inhibits tumor growth will reduce the rate of tumor growth compared to the rate of tumor growth in the absence of the TCR or composition or cell.

[0212] An "effective amount" of an agent, e.g., a pharmaceutical formulation, TCR, cell, or composition, refers to an amount effective, in the context of administration, at dosages / amounts and for periods of time necessary to achieve a desired result, such as a therapeutic or prophylactic result.

[0213] A "therapeutically effective amount" of an agent, e.g., a pharmaceutical formulation, TCR, or cell, refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result, such as treatment of a disease, condition, or disorder, and / or the pharmacokinetic or pharmacodynamic effects of treatment. A therapeutically effective amount may vary according to factors such as the disease state, age, sex, and weight of the subject, and the population of cells administered. In some embodiments, provided methods involve administering an effective amount, e.g., a therapeutically effective amount, of a TCR, cell, and / or composition.

[0214] A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, though not necessarily, the prophylactically effective amount is less than the therapeutically effective amount, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease.

[0215] As used herein, a "subject" is a mammal (eg, a human or other animal), typically a human.

[0216] Among the diseases to be treated is cancer. In some embodiments, the disease or condition to be treated is a liquid tumor. In some embodiments, the disease or condition to be treated is a hematopoietic tumor. In some embodiments, the disease or condition to be treated is lymphoma. In some embodiments, the disease or condition to be treated is acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), or acute lymphoblastic leukemia (ALL). In some embodiments, the disease or condition to be treated is chronic myeloid leukemia (CML).

[0217] In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs may be used to treat autoimmune diseases.

[0218] In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs may be used to treat genetic disorders of blood cells, which may be, but are not limited to, thalassemia and hemoglobinopathies.

[0219] In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs may be used in stem cell replacement. In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs may be used to promote the establishment of tolerance to solid organ transplants.

[0220] In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs can be used to facilitate bone marrow engraftment with lower and less toxic doses of radiation and chemotherapy for the recipient.

[0221] In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs can be used to promote lower intensity conditioning regimens alloSCT. In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs can be used to promote lower intensity conditioning regimens alloSCT for use in treating autoimmune diseases or genetic disorders of blood cells (e.g., thalassemia or hemoglobinopathies).

[0222] A. Exemplary Hematologic Malignancies, Allogeneic Stem Cell Transplantation 1. Acute myeloid leukemia (AML) Although AML is the most common indication for alloSCT, only half of patients with early- to intermediate-stage disease and one-third of patients with advanced disease survived 3 years after transplantation. See D'Souza et al. (2020) Biol Blood Marrow Transplant J Am Soc Blood Marrow Transplant 26(8):e177-e182. The most common cause of death in both early and late stages is relapse of the underlying disease. Recipients with overt active AML (i.e., morphologically evident disease >5% in the bone marrow) or measurable residual disease (MRD) at the time of alloSCT have a worse post-transplant prognosis than patients without MRD at the time of alloSCT. Methods for determining the presence or absence of MRD have evolved significantly and include assessment of morphological remission, multiparameter flow cytometry (MFC), and next-generation sequencing (NGS). MFC and NGS are based on morphology and show a 1:20 to 1 x 10 4 ~1:10 6This allows the presence of MRD to be determined down to the cell level. See Schuurhuis et al. (2018) Blood 131(12):1275-1291 and Getta et al. (2017) Biol Blood Marrow Transpl. 23(7):1064-1071.

[0223] Long-term outcomes for patients with MRD are poor, with relapse occurring in 65% of subjects, resulting in a 13% RFS and 19-23% overall survival (OS) at 3 years. See Araki et al. (2016) J Clin Oncol. 34(4):329-336 and Duval et al. (2010) J Clin Oncol. 28(23):3730-3738. The 3-year relapse rate in one retrospective study of MRD-positive patients was 67% (similar to the 65% relapse rate found in patients with active AML), compared with 22% relapse in patients with MRD-negative remission. See Araki et al. (2016) J Clin Oncol. 34(4):329-336. Other published studies have yielded similar results. See Mohty et al. (2017) Haematologica. 102(1):184-191, Decroocq et al. (2018) Am J Hematol. 93(3):416-423, and Walter et al. (2013) Blood 122(10):1813-1821. Despite very poor outcomes, one retrospective study demonstrated a benefit of alloSCT for MRD-positive patients compared with chemotherapy as a transplant-free option. See Jurjen et al. (2017) JCO Precis Oncol. (1):1-13.

[0224] Therapies targeting specific mutations, such as IDH or FLT3 inhibitors, are used in salvage regimens and are increasingly being used upfront. See Lai et al. (2019) J Hematol Oncol 12(1):100. However, despite these agents being active in only a minority of leukemias, relapsed / refractory disease remains a major clinical problem. Similarly, post-transplant hypomethylating agents are currently used, but their effect on long-term survival is uncertain. See Platzbecker et al. (2012) Leukemia 26(3):381-389, Craddock et al. (2019) J Clin Oncol Off J Am Soc Clin Oncol. 37(7):580-588, and Rautenberg et al. (2020) Bone Marrow Transplant 1-9.

[0225] AlloSCT is used for AML with active disease or MRD, even though 60% of recipients relapse within the first year and fewer than one-third of patients become long-term survivors. See D'Souza et al. (Biol Blood Marrow Transplant J Am Soc Blood Marrow Transplant 2020, 26(8):e177-e182). Most patients with overt disease are typically not offered alloSCT due to this likelihood of relapse during the first year. There is an urgent unmet medical need to extend the length of time to relapse (e.g., extend the time to relapse beyond 1, 2, 3, 4, or 5 years) in MRD-positive patients who undergo alloSCT. There is also an urgent unmet medical need to prevent relapse in MRD-positive patients who undergo alloSCT. In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs provide improved treatment options for these patients.

[0226] 2. Myelodysplastic syndrome The risk of MDS recurrence after alloSCT is higher in patients transplanted with higher-risk disease, as measured by the Revised International Prognostic Scoring System (IPSS-R). Approximately 50–60% of patients with very poor-risk MDS relapsed 2 years after alloSCT. Monosomic cytogenetic abnormalities are also associated with a higher risk of relapse, independent of IPSS score. See Koenecke et al. (2015) Haematologica. 100(3):400–408 and Deeg et al. (2012) Blood 120(7):1398–1408. More recently, specific somatic mutation profiles have been shown to predict MDS recurrence after alloSCT. Pretransplant TP53 mutations were associated with a very poor outcome, with a 3-year overall survival of less than 20% and a median survival of 0.7 years. See Lindsley et al. (2017) N Engl J Med. 376(6):536-547 and Ciurea et al. (2018) Blood 131(26):2989-2992. Other mutations related to the RAS pathway, JAK2, RUNX1, and ASXL1, have also been associated with poor outcomes after alloSCT. See Lindsley et al. (2017) N Engl J Med. 376(6):536-547 and Della Porta et al. (2016) J Clin Oncol. 34(30):3627-3637.

[0227] 3. Acute lymphoblastic leukemia (ALL) AlloSCT for ALL with active disease or primary induction failure achieved only 16% long-term survival at 3 years, as 41% of patients died from relapsed ALL before 6 months. See Duval et al. (2010) J Clin Oncol. 28(23):3730-3738.

[0228] 4. Chronic myeloid leukemia (CML) In some embodiments, TCRs and / or engineered T cells expressing TCRs may be used to treat CML, hi some embodiments, TCRs and / or engineered T cells expressing TCRs may be used to treat CML in subjects who have undergone bone marrow transplantation (e.g., SCT) or in subjects with blast crisis or accelerated phase blast crisis.

[0229] 5. Additional Diseases and Conditions Additional diseases or conditions treated using the described TCRs, T cells, and methods, including, but not limited to, liquid tumors, hematopoietic tumors, and lymphomas.

[0230] In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs can be used in bone marrow transplants for autoimmune disorders, solid tumor treatment, immune system replacement, and the like. The described T cells may also be combined with other adoptive cell therapies targeting solid tumors or autoimmune diseases or conditions. The described T cells can be used as a pre- or concurrent therapy or additive to reduce, inhibit, or eliminate the recipient's natural immune response or immune cells.

[0231] In some embodiments, the described TCRs can be isolated as soluble TCRs and administered to a subject. In some embodiments, the described TCRs can be isolated and conjugated to a molecule, such as a therapeutic molecule or antibody. The conjugated TCR can then be administered to a subject.

[0232] 6. Immune Cell Depletion In some embodiments, the described TCR and / or engineered T cells expressing the described TCR are administered to a subject to deplete the subject's immune cells. The subject may or may not have undergone a transplant. The subject may or may not be scheduled to undergo a transplant. The subject may or may not be eligible for a transplant. In some embodiments, the described TCR and / or engineered T cells expressing the described TCR are administered to a subject before the subject undergoes a transplant. In some embodiments, the described TCR and / or engineered T cells expressing the described TCR are administered to a subject after the subject undergoes a transplant. In some embodiments, the described TCR and / or engineered T cells expressing the described TCR are administered to a subject simultaneously with administration of the transplant. In some embodiments, the described TCR and / or engineered T cells expressing the described TCR are administered to a subject who has not undergone a transplant. In some embodiments, the described TCRs and / or engineered T cells expressing the described TCRs are administered to a subject who has not undergone or is not scheduled to undergo a transplant. The transplant may be, but is not limited to, a hematopoietic transplant, a stem cell transplant, or an alloSCT. In some embodiments, the subject has cancer. The cancer may be, but is not limited to, a hematopoietic cancer. In some embodiments, the subject does not have cancer. In some embodiments, the subject has an autoimmune disorder.

[0233] B. Allogeneic Stem Cell Transplantation and the Risk of Relapse More than 9,000 alloSCTs were performed in the United States in 2019, primarily as a potentially curative treatment for patients with various hematologic malignancies. See D'Souza et al. (2020) Biol Blood Marrow Transplant J Am Soc Blood Marrow Transplant 26(8):e177-e182. Post-transplant relapse remains the leading cause of graft failure, occurring in 20-40% of standard-risk and 40-80% of high-risk patients, accounting for more than half of deaths after alloSCT. See Horowitz et al. (2018) Bone Marrow Transpl. 53(11):1379-1389. There is an urgent need to extend recurrence-free survival (RFS) through novel strategies to enhance GVL without causing severe GVHD. There is also an urgent need to prevent and treat post-transplant relapse through new strategies to enhance GVL without causing severe GVHD.

[0234] The number of relapses among currently transplanted patients likely underestimates the unmet need. An analysis of AML patients is illustrative of this point. In 2018, over 3,000 alloSCTs were performed for AML in the United States. However, during the same period, there were approximately 21,450 new cases of AML, and an estimated 11,000 deaths annually. The decision to refer a patient for alloSCT depends on the benefit of relapse control versus the risk of treatment-related mortality (TRM). If a new treatment results in a lower relapse rate without significant toxicity and increased TRM, more subjects may be referred for that treatment.

[0235] Relapse is the most common cause of death after alloSCT for all types of hematologic malignancies. Because post-transplant relapse has extremely poor outcomes, RFS or cumulative relapse can be used as a reliable surrogate endpoint for survival in alloSCT. The strongest predictor of relapse is measurable residual disease (MRD) at the time of alloSCT. Even in cases with low disease burden (i.e., <5% of the bone marrow), outcomes are as poor as in patients with overt active disease. See Araki et al. (2016) J Clin Oncol. 34(4):329-336.

[0236] C. Additional Treatment Considerations In some embodiments, although unlikely, it is possible that the miHA TCR lacks specificity or exhibits on-target / off-tumor effects. The latter may occur when the HA-2 target is sufficiently expressed in non-hematopoietic tissues. To address such occurrences, strategies and methods are provided herein that should not be construed as limiting.

[0237] If GVHD occurs after administration of the miHA TCR, standard immunosuppressive therapy is initiated. As a built-in safety mechanism, the engineered cells described herein may contain an extracellular membrane-bound marker containing the CD20 epitope. The CD20 epitope is recognized by certain antibodies, including, for example, the monoclonal antibody RITUXAN® (rituximab). Recognition of the CD20 epitope may allow for selective deletion of the engineered cells. This strategy may be employed in combination with standard therapeutic interventions to reduce GVHD.

[0238] Although cytokine release syndrome (CRS) has occurred after CAR-T cell infusion, the risk of CRS may be low with TCR cell therapy. Even so, CRS remains a possibility, especially if transduced cells are rapidly and synchronously activated. If CRS occurs, standard treatment, as defined by the American Society for Transplantation and Cellular Therapy (ASTCT) consensus grading guidelines, would be initiated. Such treatments include, for example, administration of antibodies blocking IL-6 function and / or corticosteroids. See Lee et al. (2019) Biol Blood Marrow Transpl. 25(4):625-638.

[0239] The described anti-HA-2 BiTEs and anti-HA-2 CARs can be used in the treatment of cancer. Also described are methods of treating cancer in a subject, comprising administering to the subject any of the described anti-HA-2 BiTEs or anti-HA-2 CAR T cells. The anti-HA-2 BiTEs or anti-HA-2 CAR T cells can be administered to a subject to induce an immune response, increase T-cell infiltration of tumors, reduce or inhibit cancer cell proliferation, reduce or inhibit tumor growth, reduce tumor progression, reduce tumor burden, inhibit or reduce metastasis, reduce or inhibit the development of metastatic cancer, increase or extend the survival or lifespan of the subject, or reduce or attenuate an autoimmune response.

[0240] VI. Definition Unless otherwise defined, all terminology, notation, and other technical and scientific terms or terminology used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms having commonly understood meanings are defined herein for clarity and / or ease of reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial difference from what is commonly understood in the art.

[0241] "Relatively hematopoietic restricted" with respect to an antigen such as the HA-2 antigen indicates that the antigen is expressed by cells of hematopoietic origin and that there is substantially less, little, or no expression of the antigen in most other cells of a subject.

[0242] The terms "polypeptide" and "protein" are used interchangeably to refer to polymers of amino acid residues and are not limited to a minimum length. Polypeptides, including the provided T cell receptors, antigen-binding fragments thereof, and other peptides, e.g., linkers, can contain amino acid residues, including natural and / or unnatural amino acid residues. The term also includes post-expression modifications of the polypeptide, e.g., glycosylation, sialylation, acetylation, phosphorylation, and the like. In some embodiments, a polypeptide can contain modifications relative to the native or naturally occurring sequence, so long as the protein maintains the desired activity. These modifications can be deliberate, such as through site-directed mutagenesis, or can be accidental, such as through mutations of hosts producing the protein or errors due to PCR amplification.

[0243] An "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. Isolated nucleic acid includes a nucleic acid molecule contained in a cell that ordinarily contains the nucleic acid molecule, but the nucleic acid molecule is present in an extrachromosomal or chromosomal location that is different from its natural chromosomal location.

[0244] An "isolated nucleic acid molecule encoding a TCR" refers to a single nucleic acid molecule (eg, a single vector) that encodes a TCR, such as a functional α / β TCR or a functional γ / δ TCR.

[0245] An "isolated nucleic acid molecule encoding an antigen-binding fragment of a TCR" refers to a single nucleic acid molecule (eg, a single vector) that encodes an antigen-binding fragment of a TCR.

[0246] An "isolated nucleic acid molecule encoding a TCR" refers to two or more separate nucleic acid molecules (e.g., two or more vectors) that together encode a TCR, such as a functional α / β TCR or a functional γ / δ TCR. Each of such two or more nucleic acid molecules may be present in a different location within a host cell.

[0247] An "isolated nucleic acid molecule encoding an antigen-binding fragment of a TCR" refers to two or more nucleic acid molecules (e.g., two or more vectors) that together encode an antigen-binding fragment of a TCR. Each of such two or more nucleic acid molecules can be present in a different location within a host cell.

[0248] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and its progeny, regardless of the number of passages. The progeny may not be completely identical in nucleic acid content to the parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

[0249] As used herein, "percent (%) amino acid sequence identity" and "percent identity," when used in reference to an amino acid sequence (reference polypeptide sequence), are defined as the percentage of amino acid residues in a candidate sequence (e.g., a subject T cell receptor or fragment) that are identical to the amino acid residues in the reference polypeptide sequence after aligning the sequences, introducing gaps as necessary to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in a variety of ways within the skill of the art, using publicly available computer software such as, for example, BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximum alignment over the entire length of the sequences being compared.

[0250] Amino acid substitutions may involve replacing one amino acid in a polypeptide with another. Amino acid substitutions may be introduced into a TCR or antigen-binding fragment thereof of interest and the products may be screened for a desired activity, such as retained / improved antigen binding, reduced immunogenicity, or improved cytolytic activity.

[0251] Amino acids can be broadly classified according to the following common side chain properties: (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) Acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) Aromatic: Trp, Tyr, Phe.

[0252] In some embodiments, conservative substitutions may involve exchanging a member of one of these classes for another member of the same class, while in some embodiments, non-conservative amino acid substitutions may involve exchanging a member of one of these classes for another class.

[0253] The term "plasmid" or "vector" includes any known delivery vector, including bacterial delivery vectors, viral vector delivery vectors, peptide immunotherapy delivery vectors, DNA immunotherapy delivery vectors, episomal plasmids, integrating plasmids, or phage vectors. The term "vector" refers to a construct capable of delivering and, optionally, expressing one or more fusion polypeptides in a host cell. In some embodiments, a vector is capable of propagating another nucleic acid to which it is linked. The term includes vectors as self-replicating nucleic acid structures as well as vectors integrated into the genome of a host cell into which it is introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked. Such vectors are referred to herein as "expression vectors."

[0254] As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated. For example, "a" or "an" means "at least one" or "one or more." It is understood that the aspects and variations described herein include "consisting of" and / or "consisting essentially of" aspects and variations.

[0255] Throughout this disclosure, various aspects of the claimed subject matter are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the claimed subject matter. Accordingly, the description of a range should be construed as having all possible subranges specifically disclosed as well as individual numerical values ​​within that range. For example, when a range of values ​​is provided, it is understood that each intervening value between the upper and lower limits of that range, and any other stated or intervening value within that stated range, is encompassed within the claimed subject matter. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the claimed subject matter, subject to any specifically excluded limit in the stated range. Where a stated range includes one or both of the limits, ranges excluding either one or both of those included limits are also encompassed within the claimed subject matter. This applies regardless of the breadth of the range.

[0256] The term "about" as used herein refers to a normal error range for the respective value, which is readily known to one of ordinary skill in the art. Reference herein to "about" a value or parameter includes (and describes) embodiments relating to the value or parameter itself. For example, a description of "about X" includes a description of "X."

[0257] A composition may refer to any mixture of two or more products, substances, or compounds, including cells, which may be a solution, suspension, liquid, powder, paste, aqueous, non-aqueous, or any combination thereof.

[0258] As used herein, the statement that a cell or population of cells is "positive" for a particular marker refers to the detectable presence on or within the cell of the particular marker, typically a surface marker. When referring to a surface marker, the term refers to the presence of surface expression as detected by flow cytometry, e.g., by staining with an antibody that specifically binds to the marker and detecting the antibody, where the staining is detectable by flow cytometry at a level substantially greater than that detected by performing the same procedure using an isotype-matched control under otherwise identical conditions, and / or at a level substantially similar to that for cells known to be positive for the marker, and / or at a level substantially higher than that for cells known to be negative for the marker.

[0259] As used herein, the statement that a cell or population of cells is "negative" for a particular marker refers to the absence of substantial detectable presence on or within the cell of the particular marker, typically a surface marker. When referring to a surface marker, the term refers to the absence of surface expression as detected by flow cytometry, e.g., by staining with an antibody that specifically binds to the marker and detecting the antibody, where the staining is not detectable by flow cytometry at a level substantially greater than that detected by performing the same procedure using an isotype-matched control under otherwise identical conditions, and / or at a level substantially less than that for cells known to be positive for the marker, and / or at a level substantially the same as that for cells known to be negative for the marker.

[0260] VII. Illustrative Embodiments Among the embodiments provided are the following: 1. A T cell receptor (TCR) or antigen-binding fragment thereof, comprising an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises a complementarity-determining region 3 (CDR-3) comprising SEQ ID NO: 13, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 21; or (b) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 31, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 39; or (c) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 49, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 57; or (d) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 67, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 75; or (e) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 85, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 93; or (f) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 103, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 111; or (g) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 121, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 129; or (h) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 139, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 147; or (i) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 157, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 165; or (j) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 175, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 183; or (k) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 193, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 201; or (l) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 211, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 219; or (m) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 229, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 237; or (n) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 247, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 255; or (o) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 265, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 273; or (p) A T cell receptor (TCR) or an antigen-binding fragment thereof, wherein the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 283, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 291. 2. (a) the Vα or Vγ region comprises a complementarity determining region 1 (CDR-1) comprising SEQ ID NO: 11 and a complementarity determining region 2 (CDR-2) comprising SEQ ID NO: 12, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 19 and a CDR-2 comprising SEQ ID NO: 20; or (b) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 29 and a CDR-2 comprising SEQ ID NO: 30, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 37 and a CDR-2 comprising SEQ ID NO: 38; or (c) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 47 and a CDR-2 comprising SEQ ID NO: 48, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 55 and a CDR-2 comprising SEQ ID NO: 56; or (d) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 65 and a CDR-2 comprising SEQ ID NO: 66, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 73 and a CDR-2 comprising SEQ ID NO: 74; or (e) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 83 and a CDR-2 comprising SEQ ID NO: 84, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 91 and a CDR-2 comprising SEQ ID NO: 92; or (f) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 101 and a CDR-2 comprising SEQ ID NO: 102, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 109 and a CDR-2 comprising SEQ ID NO: 110; or (g) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 119 and a CDR-2 comprising SEQ ID NO: 120, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 127 and a CDR-2 comprising SEQ ID NO: 128; or (h) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 137 and a CDR-2 comprising SEQ ID NO: 138, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 145 and a CDR-2 comprising SEQ ID NO: 146; or (i) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 155 and a CDR-2 comprising SEQ ID NO: 156, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 163 and a CDR-2 comprising SEQ ID NO: 164; or (j) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 173 and a CDR-2 comprising SEQ ID NO: 174, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 181 and a CDR-2 comprising SEQ ID NO: 182; or (k) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 191 and a CDR-2 comprising SEQ ID NO: 192, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 199 and a CDR-2 comprising SEQ ID NO: 200; or (l) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 209 and a CDR-2 comprising SEQ ID NO: 210, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 217 and a CDR-2 comprising SEQ ID NO: 218; or (m) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 227 and a CDR-2 comprising SEQ ID NO: 228, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 235 and a CDR-2 comprising SEQ ID NO: 236; or (n) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 245 and a CDR-2 comprising SEQ ID NO: 246, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 253 and a CDR-2 comprising SEQ ID NO: 254; or (o) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 263 and a CDR-2 comprising SEQ ID NO: 264, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 271 and a CDR-2 comprising SEQ ID NO: 272; or (p) A TCR or antigen-binding fragment thereof described in embodiment 1, wherein the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 281 and a CDR-2 comprising SEQ ID NO: 282, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 289 and a CDR-2 comprising SEQ ID NO: 290. 3. A T cell receptor (TCR) or antigen-binding fragment thereof, comprising an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises CDR-1 comprising SEQ ID NO: 11, CDR-2 comprising SEQ ID NO: 12, and CDR-3 comprising SEQ ID NO: 13, and the Vβ or Vδ region comprises CDR-1 comprising SEQ ID NO: 19, CDR-2 comprising SEQ ID NO: 20, and CDR-3 comprising SEQ ID NO: 21; or (b) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 29, a CDR-2 comprising SEQ ID NO: 30, and a CDR-3 comprising SEQ ID NO: 31, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 37, a CDR-2 comprising SEQ ID NO: 38, and a CDR-3 comprising SEQ ID NO: 38; or (c) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 47, a CDR-2 comprising SEQ ID NO: 48, and a CDR-3 comprising SEQ ID NO: 49, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 55, a CDR-2 comprising SEQ ID NO: 56, and a CDR-3 comprising SEQ ID NO: 57; or (d) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 65, a CDR-2 comprising SEQ ID NO: 66, and a CDR-3 comprising SEQ ID NO: 67, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 73, a CDR-2 comprising SEQ ID NO: 74, and a CDR-3 comprising SEQ ID NO: 75; or (e) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 83, a CDR-2 comprising SEQ ID NO: 84, and a CDR-3 comprising SEQ ID NO: 85, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 91, a CDR-2 comprising SEQ ID NO: 92, and a CDR-3 comprising SEQ ID NO: 93; or (f) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 101, a CDR-2 comprising SEQ ID NO: 102, and a CDR-3 comprising SEQ ID NO: 103, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 109, a CDR-2 comprising SEQ ID NO: 110, and a CDR-3 comprising SEQ ID NO: 111; or (g) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 119, a CDR-2 comprising SEQ ID NO: 120, and a CDR-3 comprising SEQ ID NO: 121, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 127, a CDR-2 comprising SEQ ID NO: 128, and a CDR-3 comprising SEQ ID NO: 129; or (h) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 137, a CDR-2 comprising SEQ ID NO: 138, and a CDR-3 comprising SEQ ID NO: 139, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 145, a CDR-2 comprising SEQ ID NO: 146, and a CDR-3 comprising SEQ ID NO: 147; or (i) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 155, a CDR-2 comprising SEQ ID NO: 156, and a CDR-3 comprising SEQ ID NO: 157, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 163, a CDR-2 comprising SEQ ID NO: 164, and a CDR-3 comprising SEQ ID NO: 165; or (j) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 173, a CDR-2 comprising SEQ ID NO: 174, and a CDR-3 comprising SEQ ID NO: 175, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 181, a CDR-2 comprising SEQ ID NO: 182, and a CDR-3 comprising SEQ ID NO: 183; or (k) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 191, a CDR-2 comprising SEQ ID NO: 192, and a CDR-3 comprising SEQ ID NO: 193, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 199, a CDR-2 comprising SEQ ID NO: 200, and a CDR-3 comprising SEQ ID NO: 201; or (l) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 209, a CDR-2 comprising SEQ ID NO: 210, and a CDR-3 comprising SEQ ID NO: 211, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 217, a CDR-2 comprising SEQ ID NO: 218, and a CDR-3 comprising SEQ ID NO: 219; or (m) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 227, a CDR-2 comprising SEQ ID NO: 228, and a CDR-3 comprising SEQ ID NO: 229, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 235, a CDR-2 comprising SEQ ID NO: 236, and a CDR-3 comprising SEQ ID NO: 237; or (n) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 245, a CDR-2 comprising SEQ ID NO: 246, and a CDR-3 comprising SEQ ID NO: 247, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 253, a CDR-2 comprising SEQ ID NO: 254, and a CDR-3 comprising SEQ ID NO: 255; or (o) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 263, a CDR-2 comprising SEQ ID NO: 264, and a CDR-3 comprising SEQ ID NO: 265, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 271, a CDR-2 comprising SEQ ID NO: 272, and a CDR-3 comprising SEQ ID NO: 273; or (p) A T cell receptor (TCR) or an antigen-binding fragment thereof, wherein the Vα or Vγ region comprises CDR-1 comprising SEQ ID NO: 281, CDR-2 comprising SEQ ID NO: 282, and CDR-3 comprising SEQ ID NO: 283, and the Vβ or Vδ region comprises CDR-1 comprising SEQ ID NO: 289, CDR-2 comprising SEQ ID NO: 290, and CDR-3 comprising SEQ ID NO: 291. 4. A T cell receptor (TCR) or an antigen-binding fragment thereof, comprising an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 14, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 22; or (b) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 32, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 40; or (c) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 50, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 58; or (d) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 68, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 76; or (e) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 86, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 94; or (f) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 104, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 112; or (g) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 122, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 130; or (h) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 140, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 148; or (i) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 158, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 166; or (j) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 176, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 184; or (k) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence ...

Claims

1. A T cell receptor (TCR) or an antigen-binding fragment thereof, an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises a complementarity-determining region 3 (CDR-3) comprising SEQ ID NO: 13, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 21; or (b) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 31, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 39; or (c) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO:49, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO:57; or (d) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 67, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 75; or (e) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 85, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 93; or (f) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 103, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 111; or (g) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 121, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 129; or (h) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 139, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 147; or (i) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 157, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 165; or (j) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 175, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 183; or (k) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 193, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 201; or (l) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 211, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 219; or (m) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 229, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 237; or (n) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 247, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 255; or (o) the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 265, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO: 273; or (p) A T cell receptor (TCR), or an antigen-binding fragment thereof, wherein the Vα or Vγ region comprises a CDR-3 comprising SEQ ID NO: 283, and the Vβ or Vδ region comprises a CDR-3 comprising SEQ ID NO:

291.

2. (a) the Vα or Vγ region comprises a complementarity determining region 1 (CDR-1) comprising SEQ ID NO: 11 and a complementarity determining region 2 (CDR-2) comprising SEQ ID NO: 12, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 19 and a CDR-2 comprising SEQ ID NO: 20; or (b) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:29 and a CDR-2 comprising SEQ ID NO:30, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:37 and a CDR-2 comprising SEQ ID NO:38; or (c) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:47 and a CDR-2 comprising SEQ ID NO:48, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:55 and a CDR-2 comprising SEQ ID NO:56; or (d) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:65 and a CDR-2 comprising SEQ ID NO:66, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:73 and a CDR-2 comprising SEQ ID NO:74; or (e) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:83 and a CDR-2 comprising SEQ ID NO:84, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:91 and a CDR-2 comprising SEQ ID NO:92; or (f) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 101 and a CDR-2 comprising SEQ ID NO: 102, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 109 and a CDR-2 comprising SEQ ID NO: 110; or (g) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 119 and a CDR-2 comprising SEQ ID NO: 120, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 127 and a CDR-2 comprising SEQ ID NO: 128; or (h) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 137 and a CDR-2 comprising SEQ ID NO: 138, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 145 and a CDR-2 comprising SEQ ID NO: 146; or (i) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 155 and a CDR-2 comprising SEQ ID NO: 156, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 163 and a CDR-2 comprising SEQ ID NO: 164; or (j) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 173 and a CDR-2 comprising SEQ ID NO: 174, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 181 and a CDR-2 comprising SEQ ID NO: 182; or (k) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 191 and a CDR-2 comprising SEQ ID NO: 192, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 199 and a CDR-2 comprising SEQ ID NO: 200; or (l) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 209 and a CDR-2 comprising SEQ ID NO: 210, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 217 and a CDR-2 comprising SEQ ID NO: 218; or (m) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:227 and a CDR-2 comprising SEQ ID NO:228, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:235 and a CDR-2 comprising SEQ ID NO:236; or (n) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 245 and a CDR-2 comprising SEQ ID NO: 246, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 253 and a CDR-2 comprising SEQ ID NO: 254; or (o) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:263 and a CDR-2 comprising SEQ ID NO:264, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:271 and a CDR-2 comprising SEQ ID NO:272; or (p) The TCR or antigen-binding fragment thereof of claim 1, wherein the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 281 and a CDR-2 comprising SEQ ID NO: 282, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 289 and a CDR-2 comprising SEQ ID NO:

290.

3. A T cell receptor (TCR) or an antigen-binding fragment thereof, an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 11, a CDR-2 comprising SEQ ID NO: 12, and a CDR-3 comprising SEQ ID NO: 13, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 19, a CDR-2 comprising SEQ ID NO: 20, and a CDR-3 comprising SEQ ID NO: 21; or (b) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:29, a CDR-2 comprising SEQ ID NO:30, and a CDR-3 comprising SEQ ID NO:31, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:37, a CDR-2 comprising SEQ ID NO:38, and a CDR-3 comprising SEQ ID NO:38; or (c) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:47, a CDR-2 comprising SEQ ID NO:48, and a CDR-3 comprising SEQ ID NO:49, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:55, a CDR-2 comprising SEQ ID NO:56, and a CDR-3 comprising SEQ ID NO:57; or (d) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:65, a CDR-2 comprising SEQ ID NO:66, and a CDR-3 comprising SEQ ID NO:67, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:73, a CDR-2 comprising SEQ ID NO:74, and a CDR-3 comprising SEQ ID NO:75; or (e) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:83, a CDR-2 comprising SEQ ID NO:84, and a CDR-3 comprising SEQ ID NO:85, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:91, a CDR-2 comprising SEQ ID NO:92, and a CDR-3 comprising SEQ ID NO:93; or (f) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 101, a CDR-2 comprising SEQ ID NO: 102, and a CDR-3 comprising SEQ ID NO: 103, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 109, a CDR-2 comprising SEQ ID NO: 110, and a CDR-3 comprising SEQ ID NO: 111; or (g) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 119, a CDR-2 comprising SEQ ID NO: 120, and a CDR-3 comprising SEQ ID NO: 121, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 127, a CDR-2 comprising SEQ ID NO: 128, and a CDR-3 comprising SEQ ID NO: 129; or (h) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 137, a CDR-2 comprising SEQ ID NO: 138, and a CDR-3 comprising SEQ ID NO: 139, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 145, a CDR-2 comprising SEQ ID NO: 146, and a CDR-3 comprising SEQ ID NO: 147; or (i) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 155, a CDR-2 comprising SEQ ID NO: 156, and a CDR-3 comprising SEQ ID NO: 157, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 163, a CDR-2 comprising SEQ ID NO: 164, and a CDR-3 comprising SEQ ID NO: 165; or (j) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 173, a CDR-2 comprising SEQ ID NO: 174, and a CDR-3 comprising SEQ ID NO: 175, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 181, a CDR-2 comprising SEQ ID NO: 182, and a CDR-3 comprising SEQ ID NO: 183; or (k) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO: 191, a CDR-2 comprising SEQ ID NO: 192, and a CDR-3 comprising SEQ ID NO: 193, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO: 199, a CDR-2 comprising SEQ ID NO: 200, and a CDR-3 comprising SEQ ID NO: 201; or (l) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:209, a CDR-2 comprising SEQ ID NO:210, and a CDR-3 comprising SEQ ID NO:211, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:217, a CDR-2 comprising SEQ ID NO:218, and a CDR-3 comprising SEQ ID NO:219; or (m) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:227, a CDR-2 comprising SEQ ID NO:228, and a CDR-3 comprising SEQ ID NO:229, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:235, a CDR-2 comprising SEQ ID NO:236, and a CDR-3 comprising SEQ ID NO:237; or (n) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:245, a CDR-2 comprising SEQ ID NO:246, and a CDR-3 comprising SEQ ID NO:247, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:253, a CDR-2 comprising SEQ ID NO:254, and a CDR-3 comprising SEQ ID NO:255; or (o) the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:263, a CDR-2 comprising SEQ ID NO:264, and a CDR-3 comprising SEQ ID NO:265, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:271, a CDR-2 comprising SEQ ID NO:272, and a CDR-3 comprising SEQ ID NO:273; or (p) A T cell receptor (TCR) or antigen-binding fragment thereof, wherein the Vα or Vγ region comprises a CDR-1 comprising SEQ ID NO:281, a CDR-2 comprising SEQ ID NO:282, and a CDR-3 comprising SEQ ID NO:283, and the Vβ or Vδ region comprises a CDR-1 comprising SEQ ID NO:289, a CDR-2 comprising SEQ ID NO:290, and a CDR-3 comprising SEQ ID NO:

291.

4. A T cell receptor (TCR) or an antigen-binding fragment thereof, an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 14, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 22; or (b) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 32, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 40; or (c) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:50, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:58; or (d) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:68, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:76; or (e) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:86, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:94; or (f) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 104, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 112; or (g) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 122, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 130; or (h) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 140, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 148; or (i) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 158, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 166; or (j) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 176, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 184; or (k) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 194, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 202; or (l) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:212, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:220; or (m) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:230, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:238; or (n) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:248, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:256; or (o) the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:266, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:274; or (p) A T cell receptor (TCR), or an antigen-binding fragment thereof, wherein the Vα or Vγ region comprises a CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 284, and the Vβ or Vδ region comprises a CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:

292.

5. (a) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 14, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 22; or (b) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:32, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:40; or (c) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:50, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:58; or (d) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:68, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:76; or (e) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:86, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:94; or (f) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 104, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 112; or (g) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 122, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 130; or (h) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 140, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 148; or (i) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 158, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 166; or (j) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 176, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 184; or (k) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 194, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 202; or (l) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:212, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:220; or (m) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:230, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:238; or (n) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:248, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:256; or (o) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO:266, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:274; or (p) the Vα or Vγ region comprises CDR-1 and CDR-2 as contained within the Vα or Vγ region sequence of SEQ ID NO: 284, and the Vβ or Vδ region comprises CDR-1 and CDR-2 as contained within the Vβ or Vδ region sequence of SEQ ID NO:

292.

6. A T cell receptor (TCR) or an antigen-binding fragment thereof, an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 14, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 22; or (b) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:32, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:40; or (c) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:50, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:58; or (d) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:68, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:76; or (e) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:86, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:94; or (f) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 104, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 112; or (g) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 122, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 130; or (h) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 140, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 148; or (i) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 158, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 166; or (j) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 176, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 184; or (k) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO: 194, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO: 202; or (l) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:212, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:220; or (m) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:230, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:238; or (n) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:248, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:256; or (o) the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:266, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:274; or (p) A T cell receptor (TCR) or antigen-binding fragment thereof, wherein the Vα or Vγ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vα or Vγ region sequence of SEQ ID NO:284, and the Vβ or Vδ region comprises CDR-1, CDR-2, and CDR-3 as contained within the Vβ or Vδ region sequence of SEQ ID NO:

292.

7. A T cell receptor (TCR) or an antigen-binding fragment thereof, an alpha chain comprising a variable alpha (Vα) region and a beta chain comprising a variable beta (Vβ) region, or a gamma chain comprising a variable gamma (Vγ) region and a delta chain comprising a variable delta (Vδ) region; (a) the Vα or Vγ region comprises SEQ ID NO: 14, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 22, or a sequence having at least 90% sequence identity thereto; or (b) the Vα or Vγ region comprises SEQ ID NO: 32, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 40, or a sequence having at least 90% sequence identity thereto; or (c) the Vα or Vγ region comprises SEQ ID NO: 50, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 58, or a sequence having at least 90% sequence identity thereto; or (d) the Vα or Vγ region comprises SEQ ID NO: 68, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 76, or a sequence having at least 90% sequence identity thereto; (e) the Vα or Vγ region comprises SEQ ID NO:86, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO:94, or a sequence having at least 90% sequence identity thereto; (f) the Vα or Vγ region comprises SEQ ID NO: 104, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 112, or a sequence having at least 90% sequence identity thereto; (g) the Vα or Vγ region comprises SEQ ID NO: 122, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 130, or a sequence having at least 90% sequence identity thereto; (h) the Vα or Vγ region comprises SEQ ID NO: 140, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 148, or a sequence having at least 90% sequence identity thereto; (i) the Vα or Vγ region comprises SEQ ID NO: 158, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 166, or a sequence having at least 90% sequence identity thereto; or (j) the Vα or Vγ region comprises SEQ ID NO: 176, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 184, or a sequence having at least 90% sequence identity thereto; (k) the Vα or Vγ region comprises SEQ ID NO: 194, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 202, or a sequence having at least 90% sequence identity thereto; (l) the Vα or Vγ region comprises SEQ ID NO:212, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO:220, or a sequence having at least 90% sequence identity thereto; (m) the Vα or Vγ region comprises SEQ ID NO:230, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO:238, or a sequence having at least 90% sequence identity thereto; (n) the Vα or Vγ region comprises SEQ ID NO:248, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO:256, or a sequence having at least 90% sequence identity thereto; (o) the Vα or Vγ region comprises SEQ ID NO: 266, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 274, or a sequence having at least 90% sequence identity thereto; or (p) A T cell receptor (TCR), or an antigen-binding fragment thereof, wherein the Vα or Vγ region comprises SEQ ID NO: 284, or a sequence having at least 90% sequence identity thereto, and the Vβ or Vδ region comprises SEQ ID NO: 292, or a sequence having at least 90% sequence identity thereto.

8. (a) the Vα or Vγ region comprises SEQ ID NO: 14 and the Vβ or Vδ region comprises SEQ ID NO: 22; or (b) the Vα or Vγ region comprises SEQ ID NO: 32 and the Vβ or Vδ region comprises SEQ ID NO: 40; or (c) the Vα or Vγ region comprises SEQ ID NO: 50 and the Vβ or Vδ region comprises SEQ ID NO: 58; or (d) the Vα or Vγ region comprises SEQ ID NO: 68 and the Vβ or Vδ region comprises SEQ ID NO: 76; or (e) the Vα or Vγ region comprises SEQ ID NO: 86 and the Vβ or Vδ region comprises SEQ ID NO: 94; or (f) the Vα or Vγ region comprises SEQ ID NO: 104 and the Vβ or Vδ region comprises SEQ ID NO: 112; or (g) the Vα or Vγ region comprises SEQ ID NO: 122 and the Vβ or Vδ region comprises SEQ ID NO: 130; or (h) the Vα or Vγ region comprises SEQ ID NO: 140 and the Vβ or Vδ region comprises SEQ ID NO: 148; or (i) the Vα or Vγ region comprises SEQ ID NO: 158 and the Vβ or Vδ region comprises SEQ ID NO: 166; or (j) the Vα or Vγ region comprises SEQ ID NO: 176 and the Vβ or Vδ region comprises SEQ ID NO: 184; or (k) the Vα or Vγ region comprises SEQ ID NO: 194 and the Vβ or Vδ region comprises SEQ ID NO: 202; or (l) the Vα or Vγ region comprises SEQ ID NO: 212 and the Vβ or Vδ region comprises SEQ ID NO: 220; or (m) the Vα or Vγ region comprises SEQ ID NO: 230 and the Vβ or Vδ region comprises SEQ ID NO: 238; or (n) the Vα or Vγ region comprises SEQ ID NO: 248 and the Vβ or Vδ region comprises SEQ ID NO: 256; or (o) the Vα or Vγ region comprises SEQ ID NO: 266 and the Vβ or Vδ region comprises SEQ ID NO: 274; or (p) The TCR or antigen-binding fragment thereof of any one of claims 1 to 7, wherein the Vα or Vγ region comprises SEQ ID NO: 284 and the Vβ or Vδ region comprises SEQ ID NO:

292.

9. the alpha chain further comprises an alpha constant (Cα) region and the beta chain further comprises a beta constant (Cβ) region; or 9. The TCR or antigen-binding fragment thereof of claim 1, wherein the gamma chain further comprises a gamma constant (Cγ) region and the delta chain further comprises a delta constant (Cδ) region.

10. 10. The TCR or antigen-binding fragment thereof of claim 9, wherein the Cα comprises SEQ ID NO: 3 or 5 and the Cβ comprises SEQ ID NO: 7 or 9.

11. (a) the alpha or gamma chain comprises SEQ ID NO: 17 and the beta or delta chain comprises SEQ ID NO: 25; or (b) the alpha or gamma chain comprises SEQ ID NO: 35 and the beta or delta chain comprises SEQ ID NO: 43; or (c) the alpha or gamma chain comprises SEQ ID NO:53 and the beta or delta chain comprises SEQ ID NO:61; or (d) the alpha or gamma chain comprises SEQ ID NO: 71 and the beta or delta chain comprises SEQ ID NO: 79; or (e) the alpha or gamma chain comprises SEQ ID NO:89 and the beta or delta chain comprises SEQ ID NO:97; or (f) the alpha or gamma chain comprises SEQ ID NO: 107 and the beta or delta chain comprises SEQ ID NO: 115; or (g) the alpha or gamma chain comprises SEQ ID NO: 125 and the beta or delta chain comprises SEQ ID NO: 133; or (h) the alpha or gamma chain comprises SEQ ID NO: 143 and the beta or delta chain comprises SEQ ID NO: 151; or (i) the alpha or gamma chain comprises SEQ ID NO: 161 and the beta or delta chain comprises SEQ ID NO: 169; or (j) the alpha or gamma chain comprises SEQ ID NO: 179 and the beta or delta chain comprises SEQ ID NO: 187; or (k) the alpha or gamma chain comprises SEQ ID NO: 197 and the beta or delta chain comprises SEQ ID NO: 205; or (l) the alpha or gamma chain comprises SEQ ID NO:215 and the beta or delta chain comprises SEQ ID NO:223; or (m) the alpha or gamma chain comprises SEQ ID NO:233 and the beta or delta chain comprises SEQ ID NO:241; or (n) the alpha or gamma chain comprises SEQ ID NO:251 and the beta or delta chain comprises SEQ ID NO:259; or (o) the alpha or gamma chain comprises SEQ ID NO: 269 and the beta or delta chain comprises SEQ ID NO: 277; or (p) the alpha or gamma chain comprises SEQ ID NO: 287, and the beta or delta chain comprises SEQ ID NO:

295.

12. 12. The TCR or antigen-binding fragment thereof of any one of claims 1 to 11, wherein the TCR or antigen-binding fragment thereof recognizes a peptide epitope of the minor histocompatibility antigen HA-2 in the context of an MHC molecule.

13. The TCR or antigen-binding fragment thereof of claim 12, wherein the MHC molecule is a human leukocyte antigen (HLA)-A molecule.

14. The HLA-A molecule is of the serotype HLA-A ＊ 02:

01. The TCR or antigen-binding fragment thereof of claim 13 .

15. A TCR or antigen-binding fragment thereof according to any one of claims 12 to 14, wherein the peptide epitope of HA-2 is set forth in SEQ ID NO:

1.

16. 16. The TCR or antigen-binding fragment thereof of 15, wherein the TCR or antigen-binding fragment thereof recognizes an HA-2 peptide comprising SEQ ID NO: 1 with higher affinity than an HA-2 peptide comprising SEQ ID NO:

2.

17. A polynucleotide encoding the TCR or antigen-binding fragment thereof of any one of claims 1 to 16, or the alpha chain, beta chain, gamma chain, or delta chain thereof.

18. the polynucleotide comprises a nucleotide sequence encoding the Vα region and a nucleotide sequence encoding the Vβ region, or a nucleotide sequence encoding the Vγ region and a nucleotide sequence encoding the Vδ region; (a) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 15, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 23, or a sequence having at least 90% sequence identity thereto; or (b) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:33, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:41, or a sequence having at least 90% sequence identity thereto; or (c) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:51, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:59, or a sequence having at least 90% sequence identity thereto; or (d) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:69, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:77, or a sequence having at least 90% sequence identity thereto; (e) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:87, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:95, or a sequence having at least 90% sequence identity thereto; (f) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 105, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 113, or a sequence having at least 90% sequence identity thereto; (g) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 123, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 131, or a sequence having at least 90% sequence identity thereto; (h) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 141, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 149, or a sequence having at least 90% sequence identity thereto; or (i) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 159, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 167, or a sequence having at least 90% sequence identity thereto; or (j) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 177, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 185, or a sequence having at least 90% sequence identity thereto; (k) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 195, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 203, or a sequence having at least 90% sequence identity thereto; (l) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:213, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:221, or a sequence having at least 90% sequence identity thereto; or (m) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:231, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:239, or a sequence having at least 90% sequence identity thereto; or (n) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:249, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:257, or a sequence having at least 90% sequence identity thereto; (o) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:267, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:275, or a sequence having at least 90% sequence identity thereto; or (p) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 285, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 293, or a sequence having at least 90% sequence identity thereto.

19. (a) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 16, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 24, or a sequence having at least 90% sequence identity thereto; or (b) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:34, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:42, or a sequence having at least 90% sequence identity thereto; or (c) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:52, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:60, or a sequence having at least 90% sequence identity thereto; or (d) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 70, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 78, or a sequence having at least 90% sequence identity thereto; (e) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:88, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:96, or a sequence having at least 90% sequence identity thereto; or (f) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 106, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 114, or a sequence having at least 90% sequence identity thereto; (g) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 124, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 132, or a sequence having at least 90% sequence identity thereto; (h) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 142, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 150, or a sequence having at least 90% sequence identity thereto; (i) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 160, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 168, or a sequence having at least 90% sequence identity thereto; or (j) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 178, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 186, or a sequence having at least 90% sequence identity thereto; (k) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 196, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 204, or a sequence having at least 90% sequence identity thereto; (l) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:214, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:222, or a sequence having at least 90% sequence identity thereto; (m) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:232, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:240, or a sequence having at least 90% sequence identity thereto; (n) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:250, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:258, or a sequence having at least 90% sequence identity thereto; (o) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO:268, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO:276, or a sequence having at least 90% sequence identity thereto; or (p) the nucleotide sequence encoding the Vα or Vγ region comprises SEQ ID NO: 286, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the Vβ or Vδ region comprises SEQ ID NO: 294, or a sequence having at least 90% sequence identity thereto.

20. (a) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 18, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 26, or a sequence having at least 90% sequence identity thereto; or (b) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:36, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:44, or a sequence having at least 90% sequence identity thereto; or (c) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:54, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:62, or a sequence having at least 90% sequence identity thereto; or (d) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:72, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:80, or a sequence having at least 90% sequence identity thereto; or (e) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:90, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:98, or a sequence having at least 90% sequence identity thereto; or (f) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 108, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 116, or a sequence having at least 90% sequence identity thereto; or (g) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 126, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 134, or a sequence having at least 90% sequence identity thereto; or (h) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 144, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 152, or a sequence having at least 90% sequence identity thereto; or (i) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 162, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 170, or a sequence having at least 90% sequence identity thereto; or (j) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 180, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 188, or a sequence having at least 90% sequence identity thereto; or (k) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO: 198, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO: 206, or a sequence having at least 90% sequence identity thereto; or (l) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:216, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:224, or a sequence having at least 90% sequence identity thereto; or (m) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:234, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:242, or a sequence having at least 90% sequence identity thereto; or (n) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:252, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:260, or a sequence having at least 90% sequence identity thereto; or (o) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:270, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:278, or a sequence having at least 90% sequence identity thereto; or (p) the nucleotide sequence encoding the alpha or gamma chain comprises SEQ ID NO:288, or a sequence having at least 90% sequence identity thereto, and the nucleotide sequence encoding the beta or delta chain comprises SEQ ID NO:296, or a sequence having at least 90% sequence identity thereto.

21. 21. The polynucleotide of any one of claims 17 to 20, wherein the nucleotide sequence encoding the alpha chain and the nucleotide sequence encoding the beta chain are present on the same expression vector and are expressed from a single promoter.

22. 22. The polynucleotide of any one of claims 17 to 21, wherein the nucleotide sequence encoding the alpha chain and the nucleotide sequence encoding the beta chain are separated by a peptide sequence that causes ribosomal skipping.

23. 23. The polynucleotide of claim 22, wherein the peptide that causes ribosome skipping is a P2A peptide.

24. 24. The polynucleotide of claim 23, wherein the P2A peptide comprises SEQ ID NO:

309.

25. 25. The polynucleotide of claim 24, wherein the sequence encoding the P2A peptide is set forth in SEQ ID NO:

310.

26. 26. The polynucleotide of any one of claims 17 to 25, wherein the polynucleotide encodes the amino acid sequence of SEQ ID NO: 27, 45, 63, 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, 297, 314, 316, or 318.

27. 27. The polynucleotide of any one of claims 17 to 26, wherein the polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 28, 46, 64, 82, 100, 118, 136, 154, 172, 190, 208, 226, 244, 262, 280, 298, 315, 317, or 319.

28. A vector comprising the polynucleotide of any one of claims 17 to 27.

29. The vector of claim 28 , wherein the vector is a viral vector.

30. 30. The vector of claim 29, wherein the viral vector is a lentiviral vector.

31. 17. An engineered cell comprising the TCR or antigen-binding fragment thereof of any one of claims 1 to 16.

32. 28. An engineered cell comprising a TCR or an antigen-binding fragment thereof, wherein the TCR or antigen-binding fragment thereof is encoded by a polynucleotide according to any one of claims 17 to 27.

33. 33. The engineered cell of claim 31 or 32, wherein the TCR or antigen-binding fragment thereof is heterologous to the cell.

34. The engineered cell of any one of claims 31 to 33, wherein the engineered cell is derived from a cell line.

35. 34. The engineered cell of any one of claims 31 to 33, wherein the engineered cell is derived from a primary cell obtained from a subject.

36. 36. The engineered cell of any one of claims 31 to 35, wherein the engineered cell is a T cell, optionally wherein the T cell is a primary T cell, a natural killer T cell, or a cytotoxic T cell.

37. 37. The engineered cell of any one of claims 31 to 36, wherein expression of one or more endogenous TCR chains of said T cell is reduced or eliminated, and optionally expression of endogenous TRAC and TRBC genes of said T cell is knocked out.

38. 38. The engineered cell of any one of claims 31 to 37, wherein the engineered cell has cytotoxic activity against cells expressing the minor histocompatibility antigen (miHA) HA-2 in association with an MHC molecule.

39. 39. The engineered cell of claim 38, wherein the MHC molecule is a human leukocyte antigen (HLA)-A molecule.

40. The HLA-A molecule is of the serotype HLA-A ＊ 40. The engineered cell of claim 39, wherein the engineered cell is of the following structure: 02:

01.

41. 41. The engineered cell of any one of claims 38 to 40, wherein the HA-2 comprises SEQ ID NO:

1.

42. 42. The engineered cell of any one of claims 38 to 41, wherein the engineered cell has increased cytotoxic activity against cells expressing an HA-2 peptide comprising SEQ ID NO: 1 compared to cytotoxic activity against cells expressing an HA-2 peptide comprising SEQ ID NO:

2.

43. 31. A method for producing an engineered cell comprising introducing into a cell in vitro or ex vivo a polynucleotide according to any one of claims 17 to 27 or a vector according to any one of claims 28 to 30, optionally the cell is a T cell, optionally the T cell is a primary T cell, a natural killer T cell or a cytotoxic T cell.

44. 44. The method of claim 43, wherein the method further comprises knocking out expression of one or more endogenous TCR genes in the T cell.

45. A composition comprising the TCR or antigen-binding fragment thereof of any one of claims 1 to 16, the polynucleotide of any one of claims 17 to 27, or the vector of any one of claims 28 to 30.

46. A composition comprising the engineered cells of any one of claims 31 to 42.

47. 47. The composition of claim 45 or 46, further comprising a pharmaceutically acceptable excipient.

48. 1. A method for identifying T cell receptors (TCRs) that target the relatively hematopoietic-restricted miHA HA-2 antigen, said method comprising: (i) generating a plurality of functional TCR-encoding nucleic acid vectors by a high-throughput nucleic acid amplification and assembly method using nucleic acid obtained from a plurality of single T cells, the plurality of single T cells being from a donor subject; (ii) identifying a functional TCR that recognizes a relatively hematopoietic-restricted miHA from among the plurality of functional TCRs encoded by the plurality of functional TCR-encoding nucleic acid vectors.

49. 49. The method of claim 48, wherein the donor subject is a cancer patient, a recovered cancer patient, a stem cell transplant recipient, or a parous woman.

50. 50. The method of claim 48 or 49, wherein the donor subject does not express the relatively hematopoietic-restricted miHA HA-2 antigen, and T cells from the donor subject have been stimulated in vitro with peptides and antigen-presenting cells prior to step (i).

51. The method of any one of claims 48 to 50, wherein the identified functional TCR recognizes a peptide epitope of miHA HA-2 in the context of an MHC molecule.

52. 52. The method of claim 51, wherein the MHC molecule is a human leukocyte antigen (HLA)-A molecule.

53. The HLA-A molecule is of the serotype HLA-A ＊ 53. The method of claim 52, wherein the formula is 02:

01.

54. The method of any one of claims 48 to 53, wherein the peptide epitope of HA-2 comprises SEQ ID NO:

1.

55. 55. The method of any one of claims 48-54, wherein the T cells from the donor subject are cultured under conditions for cellular expansion of the T cells prior to the generation of the plurality of functional TCR-encoding nucleic acid vectors.

56. 55. The method of any one of claims 48-54, wherein the T cells from the donor subject are not cultured under conditions for cellular expansion of the T cells prior to said generation of the plurality of functional TCR-encoding nucleic acid vectors.

57. The high-throughput nucleic acid amplification and assembly method comprises: (1) amplifying a first amplification product and a second amplification product from complementary DNA (cDNA) generated from RNA obtained from the single T cell of the plurality of T cells sorted into each of a plurality of distinct locations on a device; amplifying the first amplification product comprising a nucleotide sequence encoding a full-length variable alpha (Vα) region or a full-length variable gamma (Vγ) region of a TCR, and the second amplification product comprising a nucleotide sequence encoding a full-length variable beta (Vβ) region or a full-length variable delta (Vδ) region of a TCR; (2) assembling the first amplification product and the second amplification product from each of the plurality of distinct locations into a nucleic acid vector to obtain an assembled nucleic acid vector comprising a nucleotide sequence encoding a functional TCR for each of the plurality of distinct locations; 57. The method of any one of claims 48-56, wherein the functional TCR comprises (i) a full-length Vα region and a full-length Vβ region derived from the single T cell, or (ii) a full-length Vγ region and a full-length Vδ region derived from the single T cell.

58. An engineered cell comprising a TCR identified by the method of any one of claims 1 to 16 or a polynucleotide of any one of claims 17 to 27.

59. The engineered cell of claim 58, wherein the engineered cell has increased cytotoxic activity against cells expressing an HA-2 peptide comprising SEQ ID NO: 1 compared to cytotoxic activity against cells expressing an HA-2 peptide comprising SEQ ID NO:

2.

60. 60. A composition comprising the engineered cells of claim 58 or 59.

61. 56. The composition of claim 55, further comprising a pharmaceutically acceptable excipient.

62. 10. A method of treatment comprising administering to a subject having a disease or disorder the composition of any one of claims 46, 47, 60 or 61, the engineered T cell of any one of claims 31-42, or the engineered T cell expressing the TCR or antigen binding fragment thereof of any one of claims 1-16 or the polynucleotide of any one of claims 17-27.

63. 1. A method of treatment, comprising administering to a subject having a disease or disorder: (a) a TCR or antigen-binding fragment according to any one of claims 1 to 16; (b) a cell expressing the TCR or antigen-binding fragment of any one of claims 1 to 16; (c) a protein comprising the TCR or antigen-binding fragment of any one of claims 1 to 16; or (d) A method of treatment comprising administering a cell expressing the protein comprising the TCR or antigen-binding fragment of any one of claims 1 to 16.

64. 64. The method of claim 62 or 63, wherein the protein comprising a TCR or antigen-binding fragment of any one of claims 1 to 16 comprises a bispecific T cell engager or a chimeric T cell receptor.

65. 65. The method of any one of claims 62-64, wherein the subject is eligible for or will undergo allogeneic hematopoietic stem cell transplantation (HSCT).

66. 66. The method of any one of claims 62 to 65, wherein the subject has or has been diagnosed with a malignant hematological disorder.

67. 67. The method of any one of claims 62-66, wherein the subject has or has been diagnosed with acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), or acute lymphoblastic leukemia (ALL).

68. 68. The method of any one of claims 62-67, wherein the subject has or has been diagnosed with a liquid tumor, a hematopoietic tumor, a lymphoma, or chronic myelogenous leukemia.

69. 69. The method of any one of claims 64-68, wherein said treating induces or enhances cell death of cells associated with said malignant hematological disorder or induces or enhances graft-versus-leukemia (GVL) in said subject.

70. 64. The method of claim 62 or 63, wherein the disease or disorder is a non-malignant disorder or an autoimmune disorder.

71. 71. The method of claim 70, wherein the autoimmune disease is a hemoglobinopathy or thalassemia.

72. 43. A method for treating a subject having a hematopoietic malignancy, comprising administering to the subject the engineered T cells of any one of claims 31-42 in combination with allogeneic stem cell transplantation (alloSCT), wherein the engineered T cells are administered to the subject within 24 hours of administering the alloSCT graft to the subject, and wherein an immunosuppressant that targets graft-versus-host disease or suppresses immune activity of T cells is not administered to the subject after administering the alloSCT graft to the subject.

73. 43. A method for treating a subject having a hematopoietic malignancy, comprising administering to the subject the engineered T cells of any one of claims 31-42 in the absence of alloSCT.

74. 43. A method for promoting the establishment of a solid organ transplant or for promoting bone marrow transplantation, the method comprising administering to a subject the engineered T cells of any one of claims 31-42.

75. 43. A method for reducing the dose of radiation or chemotherapy in a subject in need thereof for treating a hematopoietic malignancy, an autoimmune disease, or a genetic disorder of blood cells, comprising administering to the subject the engineered T cells of any one of claims 31-42.

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