T cell receptor

Abstract

The present invention provides a T cell receptor (TCR) that binds to immunogenic peptides when presented by major histocompatibility complexes (MHCs).

Description

[Technical Field]

[0001] This invention relates to T cell receptors (TCRs) that bind to immunogenic peptides when presented by major histocompatibility complexes (MHCs). The invention further relates to immunogenic peptides. [Background technology]

[0002] The fifth pillar of cancer treatment is represented by immunotherapy, which utilizes the immune system's innate ability to recognize cancer cells. In fact, the presence of tumor-infiltrating lymphocytes correlates with a better prognosis (see, for example, Zhang, L. et al., New England Journal of Medicine, 348(3), pp. 203-213).

[0003] The ability of T lymphocytes to recognize antigens is determined by the expression of T cell receptors (TCRs). TCR gene therapy is based on the gene transfer of high-avidity tumor-specific TCR genes into T lymphocytes, thus enabling specific targeting of desired tumor-associated antigens and resulting in less toxic, more specific, and effective treatment. This approach has shown promise in clinical trials.

[0004] One of the main barriers limiting the development of TCR gene therapy for the clinical treatment of cancer is the lack of tumor-specific T cells and corresponding TCRs. Tumor-specific T cells targeting tumor-associated antigens (TAAs), which are autoproteins overexpressed on cancer cells, are interesting targets in the context of adoptive T cell therapy. However, the number of isolated TAA-specific T cells remains limited, particularly in the context of hematological malignancies.

[0005] Therefore, the low availability of tumor-specific TCRs remains an unresolved challenge that limits the widespread use of TCR-based immunotherapy approaches. [Overview of the project]

[0006] The inventors identified novel TCRs that bind to immunogenic peptides when presented by major histocompatibility complexes (MHCs). They determined the amino acid sequences of these TCRs, including the amino acid sequences of their CDR regions, which are involved in their binding specificity to immunogenic peptides. These TCRs may confer tumor-specific cytolytic function to engineered T cells.

[0007] In one embodiment, the present invention relates to a T cell receptor (TCR) that binds to an immunogenic peptide when presented by a major histocompatibility complex (MHC), (1) The immunogenic peptide is a human epidermal growth factor receptor 2 (HER2) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAEGSRYGGATNKLIF (SEQ ID NO: 8) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSTFPVETQYF (SEQ ID NO: 13) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (2) The immunogenic peptide is a proteinase 3 (PR3) peptide or a neutrophil elastase (NE) peptide, and the TCR includes CDR3α containing the amino acid sequence CADYYGQNFVF (SEQ ID NO: 22) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence CASSFQGYTEAFF (SEQ ID NO: 27) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (3) The immunogenic peptide is the histone-lysine N-methyltransferase EZH2 (EZH2) peptide or the melanoma preferential expression antigen (PRAME) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CAVSESPTGFQKLVF (SEQ ID NO: 37) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α containing the amino acid sequence of CAVPHSYNTDKLIF (SEQ ID NO: 42) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CASSPRGSNTGELFF (SEQ ID NO: 47) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CSARPSEAPQYF (SEQ ID NO: 53) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (4) The immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CALPSARQLTF (SEQ ID NO: 61) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α containing the amino acid sequence of CAFMSPEGGSEKLVF (SEQ ID NO: 66) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CSVPPAGLGAPEAFF (SEQ ID NO: 71) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSPGTGAYNSPLHF (SEQ ID NO: 77) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (5) The immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAVQASNDYKLSF (SEQ ID NO: 86) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSVGRATEAFF (SEQ ID NO: 91) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (6) The immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CVVNWDNARLMF (SEQ ID NO: 98) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α containing the amino acid sequence of CATRRAKDRDDKIIF (SEQ ID NO: 459) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CASSEEGGSTDTQYF (SEQ ID NO: 103) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSLGLAGDYEQYF (SEQ ID NO: 109) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (7) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α containing the amino acid sequence CAFMKRLTQGGSEKLVF (SEQ ID NO: 119) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence CASSEVYRGHEKLFF (SEQ ID NO: 124) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (8) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAASIRSSGDKLTF (SEQ ID NO: 133) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSKRTGELFF (SEQ ID NO: 138) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (9) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CALSETLYNQGGKLIF (SEQ ID NO: 144) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSLGTSRSYTDTQYF (SEQ ID NO: 149) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (10) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CALTSDYKLSF (SEQ ID NO: 157) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CATSDLFGELFF (SEQ ID NO: 162) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASTTGIYEQYF (SEQ ID NO: 168) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (11) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAVNSWGKLQF (SEQ ID NO: 176) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASHSGLVGTGELFF (SEQ ID NO: 181) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (12) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CASTNFGNEKLTF (SEQ ID NO: 192) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSLSYEQYF (SEQ ID NO: 197) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (13) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR is a CDR3α containing (i) the amino acid sequence of CAVRSHSGNTPLVF (SEQ ID NO: 206) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) a CDR3α containing the amino acid sequence of CAENAGGTSYGKLTF (SEQ ID NO: 211) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (iii) the amino acid sequence of CAMRLPIPNNAGNMLTF (SEQ ID NO: 216) or having up to three amino acid substitutions, additions, or deletions CDR3α containing the variant thereof, and CDR3β containing (i) the amino acid sequence of CASSSPRVGPLYEQYF (SEQ ID NO: 221), or a variant thereof having up to 3 amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSDHDIYNEQFF (SEQ ID NO: 227), or a variant thereof having up to 3 amino acid substitutions, additions, or deletions, or (iii) CDR3β containing the amino acid sequence of CASSWAEEAEETQYF (SEQ ID NO: 233), or a variant thereof having up to 3 amino acid substitutions, additions, or deletions, (14) The immunogenic peptide is a Dickkopf-related protein 1 (DKK1) peptide, and the TCR comprises CDR3α containing the amino acid sequence CAVGAWYNQGGKLIF (SEQ ID NO: 241) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence CASSFGVTGELFF (SEQ ID NO: 246) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (15) The immunogenic peptide is a mucin-5AC (MUC5AC) peptide, and the TCR comprises CDR3α containing the amino acid sequence CAGGGSGAGSYQLTF (SEQ ID NO: 254) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence CAISDPTGDNQPQHF (SEQ ID NO: 259) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (16) The immunogenic peptide is a forward gradient 2 (AGR2) peptide, and the TCR comprises CDR3α containing the amino acid sequence CAVQDYGQNFVF (SEQ ID NO: 267) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence CASSPTGSEQYF (SEQ ID NO: 272) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (17) The immunogenic peptide is a c-MET peptide, and the TCR comprises CDR3α containing the amino acid sequence CAAAPNNNDMRF (SEQ ID NO: 280) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence CASSLLAGGSSYEQYF (SEQ ID NO: 285) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (18) The immunogenic peptide is a mesothelin (MSLN) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAAYNDYKLSF (SEQ ID NO: 293) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSLQGVNTEAFF (SEQ ID NO: 298) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (19) The immunogenic peptide is a CTD phosphatase subunit 1 (CTDP1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAMREGTSGTYKYIF (SEQ ID NO: 306) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSNSASRPEQYV (SEQ ID NO: 311) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (20) The immunogenic peptide is a transforming acidic coiled-coil-containing protein 2 (TACC2) peptide, and the TCR includes CDR3α containing the amino acid sequence of CAVSGLGGSNYKLTF (SEQ ID NO: 319) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASRSGTGTMDEQFF (SEQ ID NO: 324) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (21) The immunogenic peptide is a prostaglandin F2 receptor negative regulator (PTGFRN) peptide, and the TCR is provided as a T cell receptor (TCR) comprising CDR3α, which has the amino acid sequence of CAVNRDDKIIF (SEQ ID NO: 332), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which has the amino acid sequence of CSARDYKQVLRGSYNSPLHF (SEQ ID NO: 337), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0008] In one embodiment, the present invention provides a TCR that binds to the human epidermal growth factor receptor 2 (HER2) peptide when presented by MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CAEGSRYGGATNKLIF (SEQ ID NO: 8), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSTFPVETQYF (SEQ ID NO: 13), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-NSASDY (SEQ ID NO: 6), CDR2α-IRSNMDK (SEQ ID NO: 7), CDR3α-CAEGSRYGGATNKLIF (SEQ ID NO: 8), CDR1β-MNHEY (SEQ ID NO: 11), CDR2β-SVGAGI (SEQ ID NO: 12), and CDR3β-CASSTFPVETQYF (SEQ ID NO: 13), or variants thereof, each having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 9 or a variant thereof having at least 70% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 14 or a variant thereof having at least 70% sequence identity thereto. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 10 or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 15 or 16 or variants thereof having at least 70% sequence identity thereto. Preferably, the HER2 peptide comprises or consists of the amino acid sequence of KIFGSLAFL (SEQ ID NO: 5) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0009] In one embodiment, the present invention provides a TCR that binds to proteinase 3 (PR3) peptide or neutrophil elastase (NE) peptide when presented by MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence CADYYGQNFVF (SEQ ID NO: 22), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CASSFQGYTEAFF (SEQ ID NO: 27), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-VSGNPY (SEQ ID NO: 20), CDR2α-YITGDNLV (SEQ ID NO: 21), CDR3α-CADYYGQNFVF (SEQ ID NO: 22), CDR1β-MDHEN (SEQ ID NO: 25), CDR2β-SYDVKM (SEQ ID NO: 26), and CDR3β-CASSFQGYTEAFF (SEQ ID NO: 27), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 23 or a variant thereof having at least 70% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or a variant thereof having at least 70% sequence identity thereto. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 24 or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 29 or 30 or a variant thereof having at least 70% sequence identity thereto. Preferably, the PR3 peptide or NE peptide comprises or consists of the amino acid sequence of VLQELNVTV (SEQ ID NO: 19) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0010] In one embodiment, the present invention provides a TCR that, when presented by MHC, binds to the histone-lysine N-methyltransferase EZH2 (EZH2) peptide or the melanoma preferential expression antigen (PRAME) peptide, wherein the TCR comprises (i) CDR3α comprising the amino acid sequence of CAVSESPTGFQKLVF (SEQ ID NO: 37) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α comprising the amino acid sequence of CAVPHSYNTDKLIF (SEQ ID NO: 42) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β comprising the amino acid sequence of CASSPRGSNTGELFF (SEQ ID NO: 47) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β comprising the amino acid sequence of CSARPSEAPQYF (SEQ ID NO: 53) or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes the following CDR sequences: (i) CDR1α-SSVPPY (SEQ ID NO: 35), CDR2α-YTSAATLV (SEQ ID NO: 36), and CDR3α-CAVSESPTGFQKLVF (SEQ ID NO: 37), or (ii) CDR1α-VGISA (SEQ ID NO: 40), CDR2α-LSSGK (SEQ ID NO: 41), and CDR3α-CAVPHSYNTDKLIF (SEQ ID NO: 42), and (i) CDR1β-SNHLY (SEQ ID NO: 45), CDR2β-FYNNEI (SEQ ID NO: 46), and CDR3β-CASSPRGSNTGELFF (SEQ ID NO: 47), or (ii) CDR1β-DFQATT (SEQ ID NO: 51), CDR2β-SNEGSKA (SEQ ID NO: 52), and CDR3β-CSARPSEAPQYF (SEQ ID NO: 53), or variants thereof having up to three amino acid substitutions, additions, or deletions.Preferably, the TCR comprises an α-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 38 or a variant thereof having at least 70% sequence identity, or (ii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 43 or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 48 or a variant thereof having at least 70% sequence identity, or (ii) a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 54 or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing (i) the amino acid sequence of SEQ ID NO: 39, or a variant thereof having at least 70% sequence identity thereto, or (ii) an α-chain containing the amino acid sequence of SEQ ID NO: 44, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing (i) the amino acid sequence of SEQ ID NO: 49 or 50, or variants thereof having at least 70% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 55 or 56, or variants thereof having at least 70% sequence identity thereto. Preferably, the EZH2 peptide contains or consists of the amino acid sequence of KESRPPRKF (SEQ ID NO: 32), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the PRAME peptide contains or consists of the amino acid sequence of ALYVDSLFFL (SEQ ID NO: 34), or a variant thereof having up to three amino acid substitutions, additions, or deletions thereto.

[0011] In one embodiment, the present invention provides a TCR that binds to a cathepsin G (CTSG) peptide when presented by an MHC, wherein the TCR comprises (i) CDR3α comprising the amino acid sequence of CALPSARQLTF (SEQ ID NO: 61) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α comprising the amino acid sequence of CAFMSPEGGSEKLVF (SEQ ID NO: 66) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β comprising the amino acid sequence of CSVPPAGLGAPEAFF (SEQ ID NO: 71) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β comprising the amino acid sequence of CASSPGTGAYNSPLHF (SEQ ID NO: 77) or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR is one of the following CDR sequences: (i) CDR1α-ATGYPS (SEQ ID NO: 59), CDR2α-ATKADDK (SEQ ID NO: 60), and CDR3α-CALPSARQLTF (SEQ ID NO: 61), or (ii) CDR1α-TSENNYY (SEQ ID NO: 64), CDR2α-QEAYKQQN (SEQ ID NO: 65), and CDR3α-CAFMSPEGGSEKLVF (SEQ ID NO: 66), and (i) CDR1β-S (ii) QVTM (SEQ ID NO: 69), CDR2β-ANQGSEA (SEQ ID NO: 70), and CDR3β-CSVPPAGLGAPEAFF (SEQ ID NO: 71), or (ii) CDR1β-SEHNR (SEQ ID NO: 75), CDR2β-FQNEAQ (SEQ ID NO: 76), and CDR3β-CASSPGTGAYNSPLHF (SEQ ID NO: 77), or variants thereof having up to three amino acid substitutions, additions, or deletions.Preferably, the TCR comprises an α-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 62 or a variant thereof having at least 70% sequence identity, or (ii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 67 or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 72 or a variant thereof having at least 70% sequence identity, or (ii) a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 78 or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing (i) the amino acid sequence of SEQ ID NO: 63, or a variant thereof having at least 70% sequence identity thereto, or (ii) an α-chain containing the amino acid sequence of SEQ ID NO: 68, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing (i) the amino acid sequence of SEQ ID NO: 73 or 74, or a variant thereof having at least 70% sequence identity thereto, or (ii) a β-chain containing the amino acid sequence of SEQ ID NO: 79 or 80, or a variant thereof having at least 70% sequence identity thereto. Preferably, the CTSG peptide comprises or consists of a 12-mer fragment of the amino acid sequence of GIVSYGKSSGVPPEV (SEQ ID NO: 58), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0012] In one embodiment, the present invention provides a TCR that binds to a cathepsin G (CTSG) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CAVQASNDYKLSF (SEQ ID NO: 86), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSVGRATEAFF (SEQ ID NO: 91), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-VSGLRG (SEQ ID NO: 84), CDR2α-LYSAGEE (SEQ ID NO: 85), CDR3α-CAVQASNDYKLSF (SEQ ID NO: 86), CDR1β-MNHEY (SEQ ID NO: 89), CDR2β-SMNVEV (SEQ ID NO: 90), and CDR3β-CASSVGRATEAFF (SEQ ID NO: 91), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 87, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 92, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 88, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 93 or 94, or variants thereof having at least 70% sequence identity thereto. Preferably, the CTSG peptide contains or consists of the amino acid sequence of SXXXPXVFTRVSSFL (SEQ ID NO: 535), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. Preferably, the CTSG peptide contains or consists of the amino acid sequence of SGVPPEVFTRVSSFL (SEQ ID NO: 81), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CTSG peptide contains or consists of SGVPPEVFTRVSSFL (SEQ ID NO: 81), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0013] In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence VFTRVSSFL (SEQ ID NO: 82), or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence VPPEVFTRVSSFL (SEQ ID NO: 83), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0014] In one embodiment, the present invention provides a TCR that binds to a cathepsin G (CTSG) peptide when presented by an MHC, wherein the TCR comprises (i) CDR3α comprising the amino acid sequence of CVVNWDNARLMF (SEQ ID NO: 98) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α comprising the amino acid sequence of CATRRAKDRDDKIIF (SEQ ID NO: 459) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β comprising the amino acid sequence of CASSEEGGSTDTQYF (SEQ ID NO: 103) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β comprising the amino acid sequence of CASSLGLAGDYEQYF (SEQ ID NO: 109) or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR is one of the following CDR sequences: (i) CDR1α-NSASQS (SEQ ID NO: 96), CDR2α-VYSSGN (SEQ ID NO: 97), and CDR3α-CVVNWDNARLMF (SEQ ID NO: 98), or (ii) CDR1α-NSAFQY (SEQ ID NO: 457), CDR2α-TYSSGN (SEQ ID NO: 458), and CDR3α-CATRRAKDRDDKIIF (SEQ ID NO: 459), and (i) CDR1β-SN (ii) HLY (SEQ ID NO: 101), CDR2β-FYNNEI (SEQ ID NO: 102), and CDR3β-CASSEEGGSTDTQYF (SEQ ID NO: 103), or (ii) CDR1β-SGHNS (SEQ ID NO: 107), CDR2β-FNNNVP (SEQ ID NO: 108), and CDR3β-CASSLGLAGDYEQYF (SEQ ID NO: 109), or variants thereof having up to three amino acid substitutions, additions, or deletions.Preferably, the TCR comprises an α-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 99 or a variant thereof having at least 70% sequence identity, or (ii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 460 or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 104 or a variant thereof having at least 70% sequence identity, or (ii) a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 110 or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing (i) the amino acid sequence of SEQ ID NO: 100, or a variant thereof having at least 70% sequence identity thereto, or (ii) an α-chain containing the amino acid sequence of SEQ ID NO: 461, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing (i) the amino acid sequence of SEQ ID NO: 105 or 106, or variants thereof having at least 70% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 111 or 112, or variants thereof having at least 70% sequence identity thereto. Preferably, the CTSG peptide comprises or consists of a necaper fragment of the amino acid sequence of TMRSFKLLDQMETPL (SEQ ID NO: 95), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0015] In one embodiment, the present invention provides a TCR that binds to the cyclin A1 (CCNA1) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence CAFMKRLTQGGSEKLVF (SEQ ID NO: 119), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CASSEVYRGHEKLFF (SEQ ID NO: 124), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes the following CDR sequences: CDR1α-TSENNYY (SEQ ID NO: 117), CDR2α-QEAYKQQN (SEQ ID NO: 118), CDR3α-CAFMKRLTQGGSEKLVF (SEQ ID NO: 119), CDR1β-SNHLY (SEQ ID NO: 122), CDR2β-FYNNEI (SEQ ID NO: 123), and CDR3β-CASSEVYRGHEKLFF (SEQ ID NO: 124), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 120, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 125, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 121, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 126 or 127, or variants thereof having at least 70% sequence identity thereto. Preferably, the CCNA1 peptide contains or consists of a denomer fragment of the amino acid sequence of TEYAEEIYQYL (SEQ ID NO: 114), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the CTSG peptide contains or consists of the amino acid sequence of YAEEIYQYL (SEQ ID NO: 115), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0016] In one embodiment, the present invention provides a TCR that binds to the cyclin A1 (CCNA1) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CAASIRSSGDKLTF (SEQ ID NO: 133), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSKRTGELFF (SEQ ID NO: 138), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes the following CDR sequences: CDR1α-DSASNY (SEQ ID NO: 131), CDR2α-IRSNVGE (SEQ ID NO: 132), CDR3α-CAASIRSSGDKLTF (SEQ ID NO: 133), CDR1β-MDHEN (SEQ ID NO: 136), CDR2β-SYDVKM (SEQ ID NO: 137), and CDR3β-CASSKRTGELFF (SEQ ID NO: 138), or variants thereof having up to three amino acid substitutions, additions, or deletions.

[0017] Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 134, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 139, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 135, or a variant thereof having at least 70% sequence identity, and a β-chain containing the amino acid sequence of SEQ ID NO: 140 or 141, or variants thereof having at least 70% sequence identity. Preferably, the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of PETLAAFTGYSLSEI (SEQ ID NO: 128), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0018] In one embodiment, the present invention provides a TCR that binds to the cyclin A1 (CCNA1) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CALSETLYNQGGKLIF (SEQ ID NO: 144), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLGTSRSYTDTQYF (SEQ ID NO: 149), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-TRDTTYY (SEQ ID NO: 142), CDR2α-RNSFDEQN (SEQ ID NO: 143), CDR3α-CALSETLYNQGGKLIF (SEQ ID NO: 144), CDR1β-SGHTA (SEQ ID NO: 147), CDR2β-FQGNSA (SEQ ID NO: 148), and CDR3β-CASSLGTSRSYTDTQYF (SEQ ID NO: 149), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 145, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 150, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 146, or a variant thereof having at least 70% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 151 or 152, or variants thereof having at least 70% sequence identity thereto.

[0019] In one embodiment, the present invention provides a TCR that binds to the cyclin A1 (CCNA1) peptide when presented by an MHC, wherein the TCR comprises CDR3α comprising the amino acid sequence of CALTSDYKLSF (SEQ ID NO: 157) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β comprising the amino acid sequence of CATSDLFGELFF (SEQ ID NO: 162) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β comprising the amino acid sequence of CASTTGIYEQYF (SEQ ID NO: 168) or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes the following CDR sequences: CDR1α-SSNFYA (SEQ ID NO: 155), CDR2α-MTLNGDE (SEQ ID NO: 156), and CDR3α-CALTSDYKLSF (SEQ ID NO: 157), and (i) CDR1β-KGHDR (SEQ ID NO: 160), CDR2β-SFDVKD (SEQ ID NO: 161), and CDR3β-CATSDLFGELFF (SEQ ID NO: 162), or (ii) CDR1β-SNHLY (SEQ ID NO: 166), CDR2β-FYNNEI (SEQ ID NO: 167), and CDR3β-CASTTGIYEQYF (SEQ ID NO: 168), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 158, or a variant thereof having at least 70% sequence identity thereto, and a β-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 163, or a variant thereof having at least 70% sequence identity thereto, or (ii) a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 169, or a variant thereof having at least 70% sequence identity thereto. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 159, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing (i) the amino acid sequence of SEQ ID NO: 164 or 165, or variants thereof having at least 70% sequence identity thereto, or (ii) a β-chain containing the amino acid sequence of SEQ ID NO: 170 or 171, or variants thereof having at least 70% sequence identity thereto.Preferably, the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of AELSLLEADPFLKYL (SEQ ID NO: 153), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or a necaper fragment of the amino acid sequence of LLEADPFLKYLPSLI (SEQ ID NO: 527), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0020] In one embodiment, the present invention provides a TCR that binds to the cyclin A1 (CCNA1) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CAVNSWGKLQF (SEQ ID NO: 176), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASHSGLVGTGELFF (SEQ ID NO: 181), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-TSGFNG (SEQ ID NO: 174), CDR2α-NVLDGL (SEQ ID NO: 175), CDR3α-CAVNSWGKLQF (SEQ ID NO: 176), CDR1β-MNHEY (SEQ ID NO: 179), CDR2β-SMNVEV (SEQ ID NO: 180), and CDR3β-CASHSGLVGTGELFF (SEQ ID NO: 181), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 177, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 182, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 178, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 183 or 184, or variants thereof having at least 70% sequence identity thereto. Preferably, the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of WEGPGLPDFVF (SEQ ID NO: 172), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0021] In one embodiment, the present invention provides a TCR that binds to cyclin A1 (CCNA1) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CASTNFGNEKLTF (SEQ ID NO: 192), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLSYEQYF (SEQ ID NO: 197), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-TSESDYY (SEQ ID NO: 190), CDR2α-QEAYKQQN (SEQ ID NO: 191), CDR3α-CASTNFGNEKLTF (SEQ ID NO: 192), CDR1β-PRHDT (SEQ ID NO: 195), CDR2β-FYEKMQ (SEQ ID NO: 196), and CDR3β-CASSLSYEQYF (SEQ ID NO: 197), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 193, or a variant thereof having at least 70% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 198, or a variant thereof having at least 70% sequence identity thereto. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 194, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 199 or 200, or variants thereof having at least 70% sequence identity thereto. Preferably, the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of KRQLLKMEHLLLKVL (SEQ ID NO: 185), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0022] In one embodiment, the present invention provides a TCR that binds to the cyclin A1 (CCNA1) peptide when presented by an MHC, wherein the TCR is (i) a CDR3α comprising the amino acid sequence of CAVRSHSGNTPLVF (SEQ ID NO: 206), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) a CDR3α comprising the amino acid sequence of CAENAGGTSYGKLTF (SEQ ID NO: 211), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (iii) a CMRLPIPNAGNMLTF (SEQ ID NO: 216), or up to three amino acid substitutions CDR3α containing (i) the amino acid sequence of CASSSPRVGPLYEQYF (SEQ ID NO: 221), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of CASSDHDIYNEQFF (SEQ ID NO: 227), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (iii) the amino acid sequence of CASSDHDIYNEQFF (SEQ ID NO: 227), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR3β containing (iii) the amino acid sequence of CASSWAEEAEETQYF (SEQ ID NO: 233), or a variant thereof having up to three amino acid substitutions, additions, or deletions.Preferably, the TCR is one of the following CDR sequences: (i) CDR1α-TSGFYG (SEQ ID NO: 204), CDR2α-NALDGL (SEQ ID NO: 205), and CDR3α-CAVRSHSGNTPLVF (SEQ ID NO: 206), (ii) CDR1α-NSASDY (SEQ ID NO: 209), CDR2α-IRSNMDK (SEQ ID NO: 210), and CDR3α-CAENAGGTSYGKLTF (SEQ ID NO: 211), or (iii) CDR1α-TSDQSYG (SEQ ID NO: 214), CDR2α-QGSYDEQN (SEQ ID NO: 215), and CDR3α-CAMRLPIPNNAGNMLTF (SEQ ID NO: 216), and (i) CDR1β- (ii) combinations with SEHNR (SEQ ID NO: 219), CDR2β-FQNEAQ (SEQ ID NO: 220), and CDR3β-CASSSPRVGPLYEQYF (SEQ ID NO: 221), (ii) CDR1β-SEHNR (SEQ ID NO: 225), CDR2β-FQNEAQ (SEQ ID NO: 226), and CDR3β-CASSDHDIYNEQFF (SEQ ID NO: 227), or (iii) CDR1β-SNHLY (SEQ ID NO: 231), CDR2β-FYNNEI (SEQ ID NO: 232), and CDR3β-CASWAEEAEETQYF (SEQ ID NO: 233), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 207 or a variant thereof having at least 70% sequence identity, or (ii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 212 or a variant thereof having at least 70% sequence identity, or (iii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 217 or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing (i) the amino acid sequence of SEQ ID NO: 222 or a variant thereof having at least 70% sequence identity, or (ii) a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 228 or a variant thereof having at least 70% sequence identity, or (iii) a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 234 or a variant thereof having at least 70% sequence identity.Preferably, the TCR comprises an α-chain containing (i) the amino acid sequence of SEQ ID NO: 208, or a variant thereof having at least 70% sequence identity thereto, or (ii) an α-chain containing the amino acid sequence of SEQ ID NO: 213, or a variant thereof having at least 70% sequence identity thereto, or (iii) an α-chain containing the amino acid sequence of SEQ ID NO: 218, or a variant thereof having at least 70% sequence identity thereto, and (i) a β-chain containing the amino acid sequence of SEQ ID NO: 223 or 224, or a variant thereof having at least 70% sequence identity thereto, or (ii) a β-chain containing the amino acid sequence of SEQ ID NO: 229 or 230, or a variant thereof having at least 70% sequence identity thereto, or (iii) a β-chain containing the amino acid sequence of SEQ ID NO: 235 or 236, or a variant thereof having at least 70% sequence identity thereto. Preferably, the CCNA1 peptide comprises or consists of a nnomer fragment of the amino acid sequence of PETLAAFTGYSLSEI (SEQ ID NO: 201), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0023] In one embodiment, the present invention provides a TCR that binds to the Dickkopf-associated protein 1 (DKK1) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence CAVGAWYNQGGKLIF (SEQ ID NO: 241), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CASSFGVTGELFF (SEQ ID NO: 246), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-YGATPY (SEQ ID NO: 239), CDR2α-YFSGDTLV (SEQ ID NO: 240), CDR3α-YFSGDTLV (SEQ ID NO: 241), CDR1β-SGHDY (SEQ ID NO: 244), CDR2β-FNNNVP (SEQ ID NO: 245), and CDR3β-CASSFGVTGELFF (SEQ ID NO: 246), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 242, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 247, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 243, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 248 or 249, or variants thereof having at least 70% sequence identity thereto. Preferably, the DKK1 peptide comprises or consists of the amino acid sequence of ILYPGGNKV (SEQ ID NO: 238), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0024] In one embodiment, the present invention provides a TCR that binds to mucin-5AC (MUC5AC) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence CAGGGSGAGSYQLTF (SEQ ID NO: 254), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CAISDPTGDNQPQHF (SEQ ID NO: 259), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-DRGSQS (SEQ ID NO: 252), CDR2α-IYSNGD (SEQ ID NO: 253), CDR3α-CAGGGSGAGSYQLTF (SEQ ID NO: 254), CDR1β-ENHRY (SEQ ID NO: 257), CDR2β-SYGVKD (SEQ ID NO: 258), and CDR3β-CAISDPTGDNQPQHF (SEQ ID NO: 259), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 255, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 260, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 256, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 261 or 262, or variants thereof having at least 70% sequence identity thereto. Preferably, the MUC5AC peptide comprises or consists of the amino acid sequence of FLNDAGACV (SEQ ID NO: 251), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0025] In one embodiment, the present invention provides a TCR that binds to a forward gradient-2 (AGR2) peptide when presented by an MHC, wherein the TCR comprises CDR3α comprising the amino acid sequence CAVQDYGQNFVF (SEQ ID NO: 267) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β comprising the amino acid sequence CASSPTGSEQYF (SEQ ID NO: 272) or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-VSGLRG (SEQ ID NO: 265), CDR2α-LYSAGEE (SEQ ID NO: 266), CDR3α-CAVQDYGQNFVF (SEQ ID NO: 267), CDR1β-MNHEY (SEQ ID NO: 270), CDR2β-SMNVEV (SEQ ID NO: 271), and CDR3β-CASSPTGSEQYF (SEQ ID NO: 272), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 268, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 273, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 269, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 274 or 275, or variants thereof having at least 70% sequence identity thereto. Preferably, the AGR2 peptide comprises or consists of the amino acid sequence of LLVALSYTL (SEQ ID NO: 264), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0026] In one embodiment, the present invention provides a TCR that binds to the c-MET peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence CAAAPNNNDMRF (SEQ ID NO: 280), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CASSLLAGGSSYEQYF (SEQ ID NO: 285), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-DSASNY (SEQ ID NO: 278), CDR2α-IRSNVGE (SEQ ID NO: 279), CDR3α-CAAAPNNNDMRF (SEQ ID NO: 280), CDR1β-MNHEY (SEQ ID NO: 283), CDR2β-SVGAGI (SEQ ID NO: 284), and CDR3β-CASSLLAGGSSYEQYF (SEQ ID NO: 285), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 281, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 286, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 282, or a variant thereof having at least 70% sequence identity, and a β-chain containing the amino acid sequence of SEQ ID NO: 287 or 288, or variants thereof having at least 70% sequence identity. Preferably, the c-MET peptide comprises or consists of the amino acid sequence of YVDPVITSI (SEQ ID NO: 277), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0027] In one embodiment, the present invention provides a TCR that binds to a mesothelin (MSLN) peptide when presented by an MHC, wherein the TCR comprises CDR3α comprising the amino acid sequence of CAAYNDYKLSF (SEQ ID NO: 293) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β comprising the amino acid sequence of CASSLQGVNTEAFF (SEQ ID NO: 298) or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises the following CDR sequences: CDR1α-SSNFYA (SEQ ID NO: 291), CDR2α-MTLNGDE (SEQ ID NO: 292), CDR3α-CAAYNDYKLSF (SEQ ID NO: 293), CDR1β-MNHEY (SEQ ID NO: 296), CDR2β-SMNVEV (SEQ ID NO: 297), and CDR3β-CASSLQGVNTEAFF (SEQ ID NO: 298), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 294, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 299, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 295, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 300 or 301, or variants thereof having at least 70% sequence identity thereto. Preferably, the MSLN peptide comprises or consists of the amino acid sequence of KLLGPHVLGV (SEQ ID NO: 290), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0028] In one embodiment, the present invention provides a TCR that binds to the CTD phosphatase subunit 1 (CTDP1) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CAMREGTSGTYKYIF (SEQ ID NO: 306), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSNSASRPEQYV (SEQ ID NO: 311), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes the following CDR sequences: CDR1α-TSDPSYG (SEQ ID NO: 304), CDR2α-QGSYDQQN (SEQ ID NO: 305), CDR3α-CAMREGTSGTYKYIF (SEQ ID NO: 306), CDR1β-SEHNR (SEQ ID NO: 309), CDR2β-FQNEAQ (SEQ ID NO: 310), and CDR3β-CASSNSASRPEQYV (SEQ ID NO: 311), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 307, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 312, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 308, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 313 or 314, or variants thereof having at least 70% sequence identity thereto. Preferably, the CTDP1 peptide comprises or consists of the amino acid sequence of YLNKEIEEA (SEQ ID NO: 303), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0029] In one embodiment, the present invention provides a TCR that binds to a transformed acid-coiled-coil-containing protein 2 (TACC2) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CAVSGLGGSNYKLTF (SEQ ID NO: 319), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASRSGTGTMDEQFF (SEQ ID NO: 324), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes the following CDR sequences: CDR1α-SSVPPY (SEQ ID NO: 317), CDR2α-YTSAATLV (SEQ ID NO: 318), CDR3α-CAVSGLGGSNYKLTF (SEQ ID NO: 319), CDR1β-MNHNY (SEQ ID NO: 322), CDR2β-SVGAGI (SEQ ID NO: 323), and CDR3β-CASRSGTGTMDEQFF (SEQ ID NO: 324), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 320, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 325, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 321, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 326 or 327, or variants thereof having at least 70% sequence identity thereto. Preferably, the TACC2 peptide comprises or consists of the amino acid sequence of YRNSYEIEY (SEQ ID NO: 316), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0030] In one embodiment, the present invention provides a TCR that binds to a prostaglandin F2 receptor negative regulator (PTGFRN) peptide when presented by an MHC, wherein the TCR comprises CDR3α, which includes the amino acid sequence of CAVNRDDKIIF (SEQ ID NO: 332), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CSARDYKQVLRGSYNSPLHF (SEQ ID NO: 337), or a variant thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes the following CDR sequences: CDR1α-DSAIYN (SEQ ID NO: 330), CDR2α-IQSSQRE (SEQ ID NO: 331), CDR3α-CAVNRDDKIIF (SEQ ID NO: 332), CDR1β-DFQATT (SEQ ID NO: 335), CDR2β-SNEGSKA (SEQ ID NO: 336), and CDR3β-CSARDYKQVLRGSYNSPLHF (SEQ ID NO: 337), or variants thereof having up to three amino acid substitutions, additions, or deletions. Preferably, the TCR includes an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 333, or a variant thereof having at least 70% sequence identity, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 338, or a variant thereof having at least 70% sequence identity. Preferably, the TCR comprises an α-chain containing the amino acid sequence of SEQ ID NO: 334, or a variant thereof having at least 70% sequence identity thereto, and a β-chain containing the amino acid sequence of SEQ ID NO: 339 or 340, or variants thereof having at least 70% sequence identity thereto. Preferably, the PTGFRN peptide comprises or consists of the amino acid sequence of RLASRPLLL (SEQ ID NO: 329), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0031] In some embodiments, the TCR contains one or more mutations at the α-chain / β-chain interface, resulting in a reduced frequency of mispairing between the α-chain and endogenous TCR α-chain and β-chain when the α-chain and β-chain are expressed in T cells. In some embodiments, the TCR contains one or more mutations at the α-chain / β-chain interface, resulting in increased expression levels of the TCR α-chain and β-chain when the α-chain and β-chain are expressed in T cells. In some embodiments, one or more mutations introduce cysteine ​​residues into the constant region domains of the α-chain and β-chain, respectively, and the cysteine ​​residues can form disulfide bonds between the α-chain and β-chain. In some embodiments, one or more mutations are located at amino acid positions selected from those disclosed in Table 1 of Boulter, JM et al., (2003) Protein Engineering 16:707-711.

[0032] In some embodiments, the TCR includes one or more mutations to remove one or more N-glycosylation sites (see, e.g., Kuball, J et al., (2009) J Exp Med 206:463-75). Preferably, the N-glycosylation site is located in the TCR constant domain. In some embodiments, the mutation is a substitution of amino acid N to amino acid Q in the NXS / T motif. For example, the substitution may occur at one or more positions in the TCRα constant gene: positions 36, 90, or 109, and / or positions 85 and 6 in the TCRβ constant gene. In some embodiments, the substitution is located at position 36 of the TCRα constant gene.

[0033] In some embodiments, the TCR includes a mouse-like constant region.

[0034] In some embodiments, the TCR is a soluble TCR.

[0035] In another embodiment, the present invention provides polynucleotides encoding the α chain of a T cell receptor (TCR) according to the present invention, and / or the β chain of a TCR according to the present invention. In some embodiments, the polynucleotide encodes the α chain ligated to the β chain. In some embodiments, the polynucleotide further encodes one or more short interfering RNAs (siRNAs) or other agents that can reduce or prevent the expression of one or more endogenous TCR genes.

[0036] In another embodiment, the present invention provides a vector comprising a polynucleotide according to the present invention. In some embodiments, the vector is a plasmid or a viral vector. In some embodiments, the vector comprises one or more polynucleotides encoding CD3 chains, CD8, a suicide gene and / or a selection marker.

[0037] In another embodiment, the present invention provides cells comprising the TCR of the present invention, the polynucleotide of the present invention, or the vector of the present invention. In some embodiments, the cells further comprise a vector encoding one or more CD3 chains, CD8, suicide genes and / or selection markers.

[0038] In one embodiment, the present invention provides cells comprising the TCR of the present invention, the polynucleotide of the present invention, or the vector of the present invention, further comprising a vector encoding CD8. The CD8-encoding vector may be used, for example, when the TCR of the present invention is a cathepsin G-specific TCR. In some embodiments, the vector encodes a CD8α chain. In some embodiments, the vector encodes a CD8β chain. In some embodiments, the vector encodes both a CD8α chain and a CD8β chain. For example, a vector encoding both a CD8α chain and a CD8β chain may enable the expression of a CD8αβ heterodimer.

[0039] The cells of the present invention may be any suitable cells. Preferably, the cells are T cells, lymphocytes, or stem cells. In some embodiments, the cells are selected from the group consisting of CD4+ cells, CD8+ cells, naive T cells, memory stem T cells, central memory T cells, double-negative T cells, effector memory T cells, effector T cells, Th0 cells, Tc0 cells, Th1 cells, Tc1 cells, Th2 cells, Tc2 cells, Th17 cells, Th22 cells, gamma / delta T cells, natural killer (NK) cells, natural killer T (NKT) cells, cytokine-induced killer (CIK) cells, hematopoietic stem cells, and pluripotent stem cells. In some embodiments, the cells are T cells. In some embodiments, the cells are T cells isolated from the subject.

[0040] In some embodiments, endogenous genes encoding the TCRα chain and / or the TCRβ chain within the cell are preferably disrupted so that the endogenous genes encoding the TCRα chain and / or the TCRβ chain are not expressed. In some embodiments, endogenous genes encoding the TCRα chain and / or the TCRβ chain are disrupted by insertion of an expression cassette containing a polynucleotide sequence encoding the TCR of the present invention. In some embodiments, one or more endogenous genes encoding MHC are disrupted, and preferably the cells are non-alloreactive universal T cells. In some embodiments, endogenous genes involved in persistence, proliferation, activity, resistance to exhaustion / senescence / inhibitory signals, homing ability, or other T cell functions are disrupted, preferably selected from the group consisting of PD1, TIM3, LAG3, 2B4, KLRG1, TGFbR, CD160, TIGIT, CTLA4, and CD39. In some embodiments, endogenous genes involved in persistence, proliferation, activity, resistance to exhaustion / senescence / inhibitory signals, homing ability, or other T cell functions are disrupted by the incorporation of an expression cassette, the expression cassette comprising a polynucleotide sequence encoding the TCR of the present invention.

[0041] In some embodiments, endogenous genes encoding the TCRα chain and endogenous genes encoding the TCRβ chain within the cell are preferably disrupted so as not to be expressed. In some embodiments, endogenous genes encoding TIGIT are preferably disrupted so as not to be expressed. In some embodiments, endogenous genes encoding CD39 are preferably disrupted so as not to be expressed.

[0042] In some embodiments, endogenous genes encoding the TCRα chain, endogenous genes encoding the TCRβ chain, and endogenous genes encoding TIGIT within the cell are preferably disrupted so that the endogenous genes encoding the TCRα chain, endogenous genes encoding the TCRβ chain, and endogenous genes encoding TIGIT are not expressed.

[0043] In some embodiments, endogenous genes encoding the TCRα chain, endogenous genes encoding the TCRβ chain, and endogenous genes encoding CD39 within the cell are preferably disrupted so that the endogenous genes encoding the TCRα chain, endogenous genes encoding the TCRβ chain, and endogenous genes encoding CD39 are not expressed.

[0044] In some embodiments, endogenous genes encoding the TCRα chain, endogenous genes encoding the TCRβ chain, endogenous genes encoding TIGIT, and endogenous genes encoding CD39 within the cell are preferably disrupted so that the endogenous genes encoding the TCRα chain, endogenous genes encoding the TCRβ chain, endogenous genes encoding TIGIT, and endogenous genes encoding CD39 are not expressed.

[0045] In another aspect, the present invention provides T cells that are genetically engineered (e.g., gene-edited) to alter persistence, proliferation, activity, resistance to exhaustion / aging / inhibitory signals, homing ability, or other T cell functions, and the T cells express the TCRα chain and / or TCRβ chain of the present invention.

[0046] In another aspect, the present invention provides genetically engineered (e.g., gene-edited) T cells by a protocol comprising the step of targeting and incorporating expression cassettes to endogenous genes involved in persistence, proliferation, activity, resistance to exhaustion / aging / inhibitory signals, homing ability, or other T cell functions that are disrupted by artificial nucleases, wherein the expression cassettes comprise polynucleotide sequences encoding the TCRα chain and / or TCRβ chain of the present invention.

[0047] In another aspect, the present invention provides a method for preparing cells, comprising the step of introducing the polynucleotide or vector of the present invention into cells in vitro, ex vivo, or in vivo, for example, by transfection or transduction.

[0048] In another aspect, the present invention provides a method for preparing cells, comprising the step of transducing cells in vitro, ex vivo, or in vivo with one or more vectors of the present invention.

[0049] In some embodiments, the cells to be transfected or transfected are selected from the group consisting of T cells, lymphocytes, or stem cells such as hematopoietic stem cells or induced pluripotent stem cells (iPS). In some embodiments, the cells to be transfected or transfected are selected from the group consisting of CD4+ cells, CD8+ cells, Th0 cells, Tc0 cells, Th1 cells, Tc1 cells, Th2 cells, Tc2 cells, Th17 cells, Th22 cells, gamma / delta T cells, natural killer (NK) cells, natural killer T (NKT) cells, double-negative T cells, naive T cells, memory stem T cells, central memory T cells, effector memory T cells, effector T cells, cytokine-induced killer (CIK) cells, hematopoietic stem cells, and pluripotent stem cells.

[0050] In some embodiments, the method for preparing cells includes a T-cell editing step comprising disrupting an endogenous gene encoding the TCRα chain and / or an endogenous gene encoding the TCRβ chain with an artificial nuclease, which optionally includes a selection of zinc finger nucleases (ZFNs), activator-like effector nucleases (TALENs), and CRISPR / Cas systems.

[0051] In some embodiments, the method for preparing cells includes the step of targeting and incorporating an expression cassette into an endogenous gene encoding a TCRα chain and / or an endogenous gene encoding a TCRβ chain that has been disrupted by an artificial nuclease, wherein the expression cassette comprises a polynucleotide sequence encoding the TCR of the present invention or a polynucleotide sequence of the present invention.

[0052] In some embodiments, the method for preparing the cells includes a step of disrupting one or more endogenous genes encoding MHC, and preferably the cells prepared by this method are non-alloreactive universal T cells.

[0053] In some embodiments, a method for preparing cells includes disrupting one or more endogenous genes to modify persistence, proliferation, activity, resistance to exhaustion / senescence / inhibitory signals, homing ability, or other T cell functions, and optionally, the method includes targeting and incorporating an expression cassette to an endogenous gene involved in persistence, proliferation, activity, resistance to exhaustion / senescence / inhibitory signals, homing ability, or other T cell functions disrupted by an artificial nuclease, wherein the expression cassette comprises a polynucleotide sequence encoding the TCR of the present invention, and preferably, the endogenous gene is selected from the group consisting of PD1, TIM3, LAG3, 2B4, KLRG1, TGFbR, CD160, TIGIT, CTLA4, and CD39.

[0054] In some embodiments, the method for preparing cells includes the step of disrupting the endogenous gene encoding the TCRα chain and the endogenous gene encoding the TCRβ chain. In some embodiments, the method for preparing cells includes the step of disrupting the endogenous TIGIT gene. In some embodiments, the method for preparing cells includes the steps of disrupting the endogenous gene encoding the TCRα chain, the endogenous gene encoding the TCRβ chain, and the endogenous TIGIT gene. In some embodiments, the method for preparing cells includes the step of disrupting the endogenous CD39 gene. In some embodiments, the method for preparing cells includes the steps of disrupting the endogenous gene encoding the TCRα chain, the endogenous gene encoding the TCRβ chain, and the endogenous CD39 gene. In some embodiments, the method for preparing cells includes the steps of disrupting the endogenous gene encoding the TCRα chain, the endogenous gene encoding the TCRβ chain, the endogenous TIGIT gene, and the endogenous CD39 gene.

[0055] In some embodiments, two endogenous genes encoding the TCRβ chain (e.g., TRBC1 and TRBC2) are disrupted.

[0056] In another embodiment, the present invention provides a chimeric molecule comprising the TCR of the present invention or a portion thereof conjugated to a noncellular substrate, a toxin, and / or an antibody. In some embodiments, the noncellular substrate is selected from the group consisting of nanoparticles, exosomes, and other noncellular substrates.

[0057] In another aspect, the present invention provides a pharmaceutical composition comprising the TCR of the present invention, the polynucleotide of the present invention, the vector of the present invention, the cells of the present invention, cells prepared by the method of the present invention, or the chimeric molecule of the present invention.

[0058] In another aspect, the present invention provides cells of the present invention or cells prepared by the method of the present invention for use in adoptive cell transfer, preferably adoptive T cell transfer, wherein the adoptive T cell transfer is optionally allogeneic adoptive T cell transfer, autologous adoptive T cell transfer, or universal non-alloreactive adoptive T cell transfer.

[0059] In another aspect, the present invention provides the TCR of the present invention, the polynucleotide of the present invention, the vector of the present invention, the cells of the present invention, the cells prepared by the method of the present invention, the chimeric molecule of the present invention, or the pharmaceutical composition of the present invention for use in therapy.

[0060] In another aspect, the present invention provides TCRs, polynucleotides, vectors, cells, cells prepared by the methods of the present invention, chimeric molecules, or pharmaceutical compositions of the present invention for use in the treatment and / or prevention of proliferative disorders.

[0061] In another aspect, the present invention provides a method for treating and / or preventing proliferative disorders, comprising the step of administering the TCR of the present invention, the polynucleotide of the present invention, the vector of the present invention, the cells of the present invention, the cells prepared by the method of the present invention, the chimeric molecule of the present invention, or the pharmaceutical composition of the present invention to a subject in need thereof.

[0062] Proliferative disorders may be hematological malignancies or solid tumors. Preferably, hematological malignancies are selected from the group consisting of acute myeloid leukemia (AML), chronic myeloid leukemia (CML), lymphoblastic leukemia, acute lymphoblastic leukemia (ALL), myelodysplastic syndrome, lymphoma, multiple myeloma, non-Hodgkin lymphoma, and Hodgkin lymphoma. Preferably, solid tumors are selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, colon cancer, bile duct cancer, pancreatic cancer, ovarian cancer, head and neck cancer, synovial sarcoma, angiosarcoma, osteosarcoma, thyroid cancer, oral cancer, hepatocellular carcinoma, bladder cancer, endometrial cancer, neuroblastoma, rhabdomyosarcoma, liver cancer, melanoma, prostate cancer, kidney cancer, soft tissue sarcoma, urothelial carcinoma, bile duct cancer, glioblastoma, cervical cancer, mesothelioma, and colorectal cancer. In some embodiments, the proliferative disorder is acute myeloid leukemia (AML). In some embodiments, the proliferative disorder is colorectal cancer and / or pancreatic cancer.

[0063] In another embodiment, the present invention provides immunogenic cathepsin G (CTSG) peptides comprising or consisting of amino acid sequences selected from: GIVSYGKSSGVPPEV (SEQ ID NO: 58), SXXXPXVFTRVSSFL (SEQ ID NO: 535), SGVPPEVFTRVSSFL (SEQ ID NO: 81), TMRSFKLLDQMETPL (SEQ ID NO: 95), and variants thereof having up to three amino acid substitutions, additions, or deletions, or fragments thereof. Preferably, the fragments have a length of 9 amino acids or more. In some embodiments, the fragments are 9-mer fragments.

[0064] In some embodiments, the CTSG peptide comprises or consists of GIVSYGKSSGVPPEV (SEQ ID NO: 58), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof.

[0065] In some embodiments, the CTSG peptide comprises or consists of SXXXPXVFTRVSSFL (SEQ ID NO: 535), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence SGVPPEVFTRVSSFL (SEQ ID NO: 81), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence VFTRVSSFL (SEQ ID NO: 82), or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence VPPEVFTRVSSFL (SEQ ID NO: 83), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0066] In some embodiments, the CTSG peptide comprises or consists of TMRSFKLLDQMETPL (SEQ ID NO: 95), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof.

[0067] In another embodiment, the present invention provides an immunogenic cyclin A1 (CCNA1) peptide comprising or consisting of an amino acid sequence selected from the following: TEYAEEIYQYL (SEQ ID NO: 114), PETLAAFTGYSLSEI (SEQ ID NO: 128), AELSLLEADPFLKYL (SEQ ID NO: 153), LLEADPFLKYLPSLI (SEQ ID NO: 527), WEGPGLPDFVF (SEQ ID NO: 172), KRQLLKMEHLLLKVL (SEQ ID NO: 185), PETLAAFTGYSLSEI (SEQ ID NO: 201), and variants thereof having up to three amino acid substitutions, additions, or deletions, or fragments thereof. Preferably, the fragment has a length of 9 amino acids or more. In some embodiments, the fragment is a 9-mer fragment.

[0068] In some embodiments, the CCNA1 peptide comprises or consists of TEYAEEIYQYL (SEQ ID NO: 114), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CCNA1 peptide comprises or consists of the amino acid sequence YAEEIYQYL (SEQ ID NO: 115), EYAEEIYQY (SEQ ID NO: 116), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0069] In some embodiments, the CCNA1 peptide comprises or consists of PETLAAFTGYSLSEI (SEQ ID NO: 128), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CCNA1 peptide comprises or consists of the amino acid sequences ETLAAFTGY (SEQ ID NO: 129), LAAFTGYSL (SEQ ID NO: 130), or variants thereof having up to three amino acid substitutions, additions, or deletions.

[0070] In some embodiments, the CCNA1 peptide comprises or consists of AELSLLEADPFLKYL (SEQ ID NO: 153), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CCNA1 peptide comprises or consists of LLEADPFLKYLPSLI (SEQ ID NO: 527), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CCNA1 peptide comprises or consists of the amino acid sequence of EADPFLKYL (SEQ ID NO: 154), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0071] In some embodiments, the CCNA1 peptide comprises or consists of WEGPGLPDFVF (SEQ ID NO: 172), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CCNA1 peptide comprises or consists of the amino acid sequence of GPGLPDFVF (SEQ ID NO: 173), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0072] In some embodiments, the CCNA1 peptide comprises or consists of KRQLLKMEHLLLKVL (SEQ ID NO: 185), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CCNA1 peptide comprises or consists of the amino acid sequences RQLLKMEHL (SEQ ID NO: 186), MEHLLLKVL (SEQ ID NO: 187), MEHLLLKV (SEQ ID NO: 188), KMEHLLLKV (SEQ ID NO: 189), or variants thereof having up to three amino acid substitutions, additions, or deletions.

[0073] In some embodiments, the CCNA1 peptide comprises or consists of PETLAAFTGYSLSEI (SEQ ID NO: 201), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CCNA1 peptide comprises or consists of the amino acid sequences of ETLAAFTGY (SEQ ID NO: 202), LAAFTGYSL (SEQ ID NO: 203), or variants thereof having up to three amino acid substitutions, additions, or deletions.

[0074] In another embodiment, the present invention provides a TCR that binds to an immunogenic CTSG peptide when presented by a major histocompatibility complex (MHC), wherein the immunogenic CTSG peptide comprises or consists of SXXXPXVFTRVSSFL (SEQ ID NO: 535), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof.

[0075] In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence SGVPPEVFTRVSSFL (SEQ ID NO: 81), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence VFTRVSSFL (SEQ ID NO: 82), or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence VPPEVFTRVSSFL (SEQ ID NO: 83), or a variant thereof having up to three amino acid substitutions, additions, or deletions. [Brief explanation of the drawing]

[0076] [Figure 1-1] Evaluation of the immune response in healthy donor T lymphocytes stimulated with 17 HLA-A*02:01 immunogenic peptides. T lymphocytes from two healthy donors (LAA-HD3 and LAA-HD5) were co-cultured for 6 hours with autologous APCs pulsed with HLA-A*02:01 immunogenic peptides derived from antigens including EZH2, PRAME, PR3, and NE, or with a pool of unrelated peptides. Antigen-specific proliferation of T lymphocytes was evaluated by flow cytometry analysis by measuring CD107a production and IFN-γ secretion. (A) The results showed higher production of CD107a and IFN-γ in response to the LAA pool for both LAA-HD3 (A, left) and LAA-HD5 (A, right) compared to unrelated samples. (B) Clonal proliferation of T lymphocytes was evaluated by flow cytometry using a kit covering 70% of the human TCR Vβ repertoire. The results showed proliferation of TRBV2, TRBV4-1, 4-2, 4-3, and TRBV28 in LAA-HD3. [Figure 1-2]Evaluation of the immune response in healthy donor T lymphocytes stimulated with 17 HLA-A*02:01 immunogenic peptides. T lymphocytes from two healthy donors (LAA-HD3 and LAA-HD5) were co-cultured for 6 hours with autologous APCs pulsed with HLA-A*02:01 immunogenic peptides derived from antigens including EZH2, PRAME, PR3, and NE, or with a pool of unrelated peptides. Antigen-specific proliferation of T lymphocytes was evaluated by flow cytometry analysis by measuring CD107a production and IFN-γ secretion. (C) and (D) LAA-T cells proliferated from the two healthy donors were characterized by TCRαβ sequencing after being stimulated several times with the peptide pool. Sequencing results showed the presence of dominant clonal types for LAA-HD3 (C) and LAA-HD5 (D). The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S4 corresponds to the sequencing result obtained after the fourth stimulation) to the TCR repertoire at each stimulus evaluated. The remaining sequences are grouped together in white. HD: healthy donor, S: stimulus, SP: subpool, APC: antigen-presenting cell, TRBV: T cell receptor β-chain variable gene, HD: healthy donor, LAA: leukemia-associated antigen, unrelated: pool of unrelated peptides, CDR3: complementarity-determining region 3, S: stimulus, TRAV: T cell receptor α-chain variable gene, TRAJ: T cell receptor α-chain ligature gene, TRBJ: T cell receptor β-chain ligature gene, TRBD: T cell receptor β-chain diversity gene. [Figure 1-3]Evaluation of the immune response in healthy donor T lymphocytes stimulated with 17 HLA-A*02:01 immunogenic peptides. T lymphocytes from two healthy donors (LAA-HD3 and LAA-HD5) were co-cultured for 6 hours with autologous APCs pulsed with HLA-A*02:01 immunogenic peptides derived from antigens including EZH2, PRAME, PR3, and NE, or with a pool of unrelated peptides. Antigen-specific proliferation of T lymphocytes was evaluated by flow cytometry analysis by measuring CD107a production and IFN-γ secretion. (C) and (D) LAA-T cells proliferated from the two healthy donors were characterized by TCRαβ sequencing after being stimulated several times with the peptide pool. Sequencing results showed the presence of dominant clonal types for LAA-HD3 (C) and LAA-HD5 (D). The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S4 corresponds to the sequencing result obtained after the fourth stimulation) to the TCR repertoire at each stimulus evaluated. The remaining sequences are grouped together in white. HD: healthy donor, S: stimulus, SP: subpool, APC: antigen-presenting cell, TRBV: T cell receptor β-chain variable gene, HD: healthy donor, LAA: leukemia-associated antigen, unrelated: pool of unrelated peptides, CDR3: complementarity-determining region 3, S: stimulus, TRAV: T cell receptor α-chain variable gene, TRAJ: T cell receptor α-chain ligature gene, TRBJ: T cell receptor β-chain ligature gene, TRBD: T cell receptor β-chain diversity gene. [Figure 2] Deconvolution of the mapping grid allows for the identification of immunogenic epitopes. Subpools capable of inducing a T cell-mediated immune response were identified via a 6-hour co-culture of T lymphocytes with self-irradiated APCs pre-pulsed with 9 SPs containing 17 immunogenic peptides. The results showed two immunogenic SPs (SP3 and SP6) for LAA-HD3 (left) and three immunogenic SPs (SP1, SP2, and SP9) for LAA-HD5 (right). HD: healthy donor, S: stimulation, SP: subpool, APC: antigen-presenting cell. [Figure 3-1]Evaluation of CTSG-specific T cell enrichment from HDs. (A) T cells isolated from two HDs (CTSG-HD1, CTSG-HD2) were co-cultured for 6 hours with autologous APCs pulsed with a CTSG peptide pool or a pool of unrelated peptides. The occurrence of a CTSG-specific response was determined by measuring CD107a expression and IFNγ secretion in T cells. The results showed time-dependent enrichment of CTSG-specific lymphocytes in HD1 (left), and in HD2 (right), 10-15% of CTSG-specific T cells were already present in the culture after three stimulations with the peptide pool. (B) Characterization of the Vβ repertoire. Clonal proliferation of CTSG-specific T cells isolated from HD1 was evaluated by flow cytometry. The results showed the dominant expression of three Vβs in HD1 (TRBV4-3, TRBV9, and TRBV27). [Figure 3-2] Evaluation of CTSG-specific T cell enrichment from HD. (C) Identification of immunogenic SPs for each HD. SPs that induce an immune response in each HD analyzed were identified by co-culturing T cells with autologous APCs pulsed with each of the 16 CTSG SPs. The results identified SP5, SP6, SP8, SP12, and SP15 for HD1 (C, left) and SP5 and SP16 for HD2 (C, right). By deconvolution of the mapping grid (C, table below), peptides 29, 30, 32, 53, 54, and 56 were identified as potential targets for HD1 (left), and peptide 61 as a potential target for HD2 (right). (D) To confirm the immunogenicity of the identified peptides, T cells were co-culturified for 6 hours with APCs pulsed with each peptide. The results show an increased response of CTSG-HD1 T cells to peptides P54 and P56. HD: Healthy donor, HLA: Human leukocyte antigen, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, CG: Cathepsin G, P: Peptide, unr: Unrelated peptide. [Figure 4]HLA restriction of CTSG-HD1 T lymphocytes. (A) A table showing the EBV cell lines used to identify the HLA restriction of CTSG-HD1 T cells. HLA elements present in EBV cells and shared with HD1 are shown in bold. (B) CTSG-HD1-derived T lymphocytes were co-cultured with EBV-BLCLs pulsed with P54, P56, or unrelated peptides, each containing one constraint element common to HD1. IFNγ production was observed for EBV-BLCL#1 (left) when T cells were cultured with P54-pulsed APCs, and for EBV-BLCL#2, #4, and #5, suggesting that T cells recognized P54 when presented on the HLA-A*02:01 allele, and P56 when presented on the HLA-A24:02 and HLA-C*07:02 constraint elements (right). HLA: Human leukocyte antigen, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, CG: Cathepsin G, P56: Peptide 56, unrel: Unrelated peptide, CTSG-HD1: Cathepsin G-healthy donor 1. [Figure 5-1] Longitudinal TCR sequencing of enriched CTSG-specific T cells. T cells generated from two healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in the CTSG pool. Sequencing results showed the presence of dominant clonal types for CTSG-HD1(A, B) and CTSG-HD2(C). For HD1, we identified two proliferated T cell populations that specifically recognized P54(A) or P56(B) in the peptide pool. The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. CDR3: Complementary Determinant Region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 5-2]Longitudinal TCR sequencing of enriched CTSG-specific T cells. T cells generated from two healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in the CTSG pool. Sequencing results showed the presence of dominant clonal types for CTSG-HD1(A, B) and CTSG-HD2(C). For HD1, we identified two proliferated T cell populations that specifically recognized P54(A) or P56(B) in the peptide pool. The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. CDR3: Complementary Determinant Region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 5-3] Longitudinal TCR sequencing of enriched CTSG-specific T cells. T cells generated from two healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in the CTSG pool. Sequencing results showed the presence of dominant clonal types for CTSG-HD1(A, B) and CTSG-HD2(C). For HD1, we identified two proliferated T cell populations that specifically recognized P54(A) or P56(B) in the peptide pool. The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. CDR3: Complementary Determinant Region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 6]Genesis and functional verification of P56-specific T cells using TCR gene transfer and TCR gene editing approaches. (A) Disruption of the endogenous TCR repertoire and CTSG-P56 TCR expression in edited T cells were evaluated by measuring CD3 surface expression in untransduced T cells or T cells transduced with LV, respectively. (B) T cell functionality was evaluated by measuring CD107a expression and IFNγ secretion in T cells during 6 hours of co-culture with EBV cell lines expressing either the HLA-A*24:02 (EBV-BLCL#6) or HLA-C*07:02 (EBV-BLCL#5) allele and pulsed with P56. Either transferred cells or TCR-edited cells were used as effector cells in the assay. (C) Phenotypic characterization of edited T cells regarding TCR disruption efficiency (C, left), P56-TCR-specific Vβ region expression (C, center), and memory phenotype (C, right). HLA: Human leukocyte antigen, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, CTSG: Cathepsin G, P56: Peptide 56, unrel: Unrelated peptide, TRBV: T cell receptor β chain variable gene, TSCM: Stem cell-like memory T cell, TCM: Central memory T cell, TEMRA: Terminal effector memory T cell, TEM: Effector memory T cell, KO: Knockout. [Figure 7-1]P56αβ-edited T cells recognized the VFT9mer on the HLA-A*24:02 allele but did not recognize the 9mer on the HLA-C*07:02 constraint element. (A) We used an online tool to predict the binding affinity of P56-derived 9mers to the HLA-A*24:02 and HLA-C*07:02 alleles. The tool predicted weak binding affinity for the VFTRVSSFL (SEQ ID NO: 82) 9mer presented by the HLA-A*24:02 molecule (left) and weak binding affinity for the same peptide presented by HLA-C07:02, but predicted that the latter would bind with higher affinity by the EVFTRVSSF (SEQ ID NO: 42) 9mer (right). (B) Edited P56αβ T cells were co-cultured for 6 hours with EBV-BLCL expressing HLA-A*24:02 or HLA-C*07:02 constraint elements and pulsed with P56-derived necapers and decamers. CD107a production in T lymphocytes was observed only in response to P7 necapers (VFTRVSSFL-SEQ ID NO: 82) presented by the HLA-A*24:02 molecule. [Figure 7-2] P56αβ-edited T cells recognized the VFT9mer on the HLA-A*24:02 allele but did not recognize the necaper on the HLA-C*07:02 constraint element. (B) Edited P56αβ T cells were co-cultured for 6 hours with EBV-BLCL expressing either HLA-A*24:02 or HLA-C*07:02 constraint elements and pulsed with P56-derived necapers and decamers. CD107a production in T lymphocytes was observed only in response to the P7 necaper (VFTRVSSFL-SEQ ID NO: 82) when presented by the HLA-A*24:02 molecule. (C) Edited P56αβ T cells were co-cultured with EBV-BLCL expressing the HLA-C*07:02 allele and pulsed with P56, P56-derived necapers and decamers. Six hours later, when T lymphocytes were co-cultured with P56-pulsed EBV cells, CD107a production was high. [Figure 7-3]P56αβ-edited T cells recognized the VFT9mer on the HLA-A*24:02 allele but did not recognize the 9mer on the HLA-C*07:02 constraint element. (D) Co-culture of edited P56αβ T lymphocytes with HLA-C*07:02 EBV-BLCL pulsed with P12, P13, and P56. CD107a expression and cytokine secretion were evaluated after 6 hours, and the results showed an increase in percentage when co-cultured with P13 and P56. (E) To determine the affinity profiles of P13 and P56 for the HLA-C07:02 allele, EBV-BLCL expressing the HLA-C07:02 allele was pulsed with either P13 or P56 alone, or in combination with a high-affinity HLA-C*07:02 binding peptide (RYRPGTVAL-SEQ ID NO: 423), and co-cultured with edited P56 TCR T cells for 6 hours. Equivalent CD107a expression and cytokine secretion were detected under conditions with P13 and P56 alone, as well as under conditions with competing substances present. Rank %: Percentage of rank, 9mer: 9-mer, P56: Peptide 56, P1: 9-mer 1, P2: 9-mer 2, P3: 9-mer 3, P4: 9-mer 4, p5: 9-mer 5, p6: 9-mer 6, p7: 9-mer 7 (VFT9-mer), p10: 10-mer, P56αβ: Transduced T lymphocytes with LV encoding P56αβ TCR, EBV-BLCL: Epstein-Barr virus transformed B lymphoblast-like cell line, 15mer (15-mer): 15-amino acid long peptide, APC: Antigen-presenting cell, P12: 12-amino acid long peptide derived from Peptide 56, P13: 13-amino acid long peptide derived from Peptide 56, CTR: High affinity HLA-C*O7:O2 binding peptide (RYRPGTVAL). [Figure 8-1] Determination of P56αβ TCR avidity when recognizing congeneral peptides on target cells. (A) P56 TCRαβ-edited T lymphocytes were co-cultured with HLA-A*24:02 EBV-BLCL pulsed with gradually decreasing concentrations of P7 nonamer (A, top), and HLA-C*07:02 EBV-BLCL pulsed with gradually decreasing concentrations of P13 (A, middle) and P56 (A, bottom). CD107a expression and cytokine secretion were evaluated by readout analysis using flow cytometry. [Figure 8-2]Determination of P56αβ TCR avidity when recognizing congeneral peptides on target cells. (A) P56 TCRαβ-edited T lymphocytes were co-cultured with HLA-A*24:02 EBV-BLCL pulsed with gradually decreasing concentrations of P7 nonamer (A, top), and HLA-C*07:02 EBV-BLCL pulsed with gradually decreasing concentrations of P13 (A, middle) and P56 (A, bottom). CD107a expression and cytokine secretion were evaluated by readout analysis using flow cytometry. [Figure 8-3] Determination of P56αβ TCR avidity when recognizing congeneral peptides on target cells. (A) P56 TCRαβ-edited T lymphocytes were co-cultured with HLA-A*24:02 EBV-BLCL pulsed with gradually decreasing concentrations of P7 nonamer (A, top), and HLA-C*07:02 EBV-BLCL pulsed with gradually decreasing concentrations of P13 (A, middle) and P56 (A, bottom). CD107a expression and cytokine secretion were evaluated by readout analysis using flow cytometry. [Figure 8-4] Determination of P56αβ TCR avidity when recognizing a congener peptide on target cells. (B) Log EC50 values ​​show a low avidity profile for P56 TCRαβ for the P7 necaper, a higher avidity for P13, but the highest avidity for P56. Unstimulated: T cells maintained without the addition of target cells, PMA / Iono: PMA and ionomycin-positive control, P7: P7 necaper, P13: 13-amino acid long peptide derived from peptide 56, TCR: T cell receptor, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, EC50: half-effect concentration (EC50). [Figure 9]Evaluation of the innate processing of immunogenic epitopes derived from peptide 56 on the surface of primary leukemic blast cells expressing only the HLA-A*24:02 allele or the HLA-C*07:02 allele. (AB) Edited P56 TCRαβ T lymphocytes were co-cultured for 24 hours with the following cells: (A) pAML1 and pAML2 (expressing only the HLA-A24:02 allele), (B) pAML3, pAML4 and pAML5 (expressing only the HLA-C07:02 allele), and (A and B) pAML6 and pAML7 (negative for both the HLA-A24:02 and HLA-C07:02 alleles) as negative controls. T lymphocytes and leukemic blast cells were plated with different E:T ratios (5:1, 1:1, 1:5). As a further negative control, corresponding numbers of leukemia cells used with different E:T ratios were also plated in the absence of T lymphocytes. The elimination index of AML blast cells was calculated using the following formula: 1 - ((count of leukemia cells with different E:T ratios (5:1, 1:1, 1:5)) / (count of the same leukemia cells alone)). A value of 0 indicates no elimination of leukemia cells, while a value of 1 indicates complete elimination of leukemia cells. Results are presented as mean standard error (SEM) (N=3). pAML, primary acute myeloid leukemia. [Figure 10] Evaluation of cyclin A1-specific T cell enrichment from HDs. (AC) T cells isolated from three HDs (cyclin A1-HD1, cyclin A1-HD2, cyclin A1-HD3) were co-cultured for 6 hours with autologous APCs pulsed with a cyclin A1 peptide pool or a pool of unrelated peptides. The occurrence of a cyclin A1-specific response was determined by measuring CD107a expression and IFNγ secretion in T cells. The results showed enrichment of tumor-specific CD8+ and CD4+ T lymphocytes for HD1(A), enrichment of cyclin A1-specific T cells for HD2(B), and progressive enrichment of cyclin A1-specific T cells for HD3(C). HD: Healthy donor. [Figure 11-1]Identification of peptide subpools that induce T cell responses. Identification of immunogenic SPs for each HD. SPs that induce immune responses in each HD analyzed were identified by co-culturing T cells with autologous APCs pulsed with each of the 22 cyclin A1 SPs. (A) For HD1, the results identify SP4, SP5, SP6, SP8, SP9, SP10, SP12, SP16, SP18, SP20 (A, top) for HD1 CD8+ T cells, and SP3 and SP21 (A, bottom) for HD1 CD4+ T cells. [Figure 11-2] Identification of peptide subpools that induce T cell responses. Identification of immunogenic SPs for each HD. SPs that induce immune responses in each HD analyzed were identified by co-culturing T cells with autologous APCs pulsed with each of 22 cyclin A1 SPs. (B) For cyclin A1 HD2 T cells, the inventors observed immune responses to SP3, SP4, and SP21. [Figure 11-3] Identification of peptide subpools that induce T cell responses. Identification of immunogenic SPs for each HD. SPs that induce immune responses in each HD analyzed were identified by co-culturing T cells with autologous APCs pulsed with each of 22 cyclin A1 SPs. (C) For HD3 cyclin A1 T cells, the inventors observed a small but still significant immune response to SP3 and SP21. [Figure 12-1] Evaluation of immunogenic peptides. To confirm the immunogenicity of the identified peptides, T cells were co-cultured for 6 hours with APCs pulsed with each peptide. The results confirmed the recognition of P102 and P52-P53 by CD4+(A) and CD8+(B) cyclin A1-HD1 T cells, respectively. T cells from passaged SP12 were co-cultured with SP12 and individual peptides belonging to SP (P4, P5, P6, P7, P10). Analysis showed recognition of P6 and P7 (C). T cells from passaged SP20 were co-cultured with SP20 and individual peptides belonging to SP (P92, P93, P94, P95, P98). Analysis showed recognition of P93 and P94 (D). [Figure 12-2] Evaluation of immunogenic peptides. To confirm the immunogenicity of the identified peptides, T cells were co-cultured for 6 hours with APCs pulsed with individual peptides. (E) Since P52 and P53 are duplicate peptides, in order to identify the exact necaper recognized by HD1 T cells, the inventors co-cultured T cells with APCs pulsed with P52, P53, the duplicate 11-mer sequence, and three individual necapers (TEY, EYA, YAE) derived from the 11-mer portion. The results of the functional assay showed CD107a expression and IFNg secretion when T cells were co-cultured with APCs pulsed with YAE necapers. (F) For cyclin A1-HD2 T cells, the inventors confirmed the recognition of SP19, and in particular, identified the immunogenic peptide P81. (G) For cyclin A1-HD3 T cells, the inventors identified peptide P102 as one that induces an immune response. HD: Healthy donor, SP: Subpool, P: Peptide [Figure 13-1] HLA restriction of cyclin A1-specific T lymphocytes. Table (A and B, left panel) shows the EBV cell lines used to identify HLA restriction of cyclin A1-HD2 and HD3 T cells. HLA elements contained in EBV cells and shared with HD2 and HD3 are highlighted. T lymphocytes derived from cyclin A1-HD2 T cells were co-cultured with EBV-BLCLs that have one restriction element common to HD2 and pulsed with P81 or an unrelated peptide. In EBV-BLCLs #3 and #7 pulsed with P81, IFNγ production and CD107a secretion were observed (A, right). For cyclin A1-HD3, the inventors observed recognition by both EBV cell lines used in the assay (B, right). HLA: Human leukocyte antigen, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, P81: Peptide 81, unrel: Unrelated peptide, HD: Healthy donor [Figure 13-2]HLA restriction of cyclin A1-specific T lymphocytes. Table (A and B, left panel) shows the EBV cell lines used to identify HLA restriction of cyclin A1-HD2 and HD3 T cells. HLA elements contained in EBV cells and shared with HD2 and HD3 are highlighted. T lymphocytes derived from cyclin A1-HD2 T cells were co-cultured with EBV-BLCLs that have one restriction element common to HD2 and pulsed with P81 or an unrelated peptide. In EBV-BLCLs #3 and #7 pulsed with P81, IFNγ production and CD107a secretion were observed (A, right). For cyclin A1-HD3, the inventors observed recognition by both EBV cell lines used in the assay (B, right). HLA: Human leukocyte antigen, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, P81: Peptide 81, unrel: Unrelated peptide, HD: Healthy donor [Figure 14-1] Cellular fluorescence immunoprofiling of T cell populations. Characterization of the Vβ repertoire. (A) Clonal proliferation of cyclin A1-specific T cells isolated from HD1, HD2, and HD3 was evaluated by flow cytometry. For HD1 T cells, analysis was performed after 5 and 7 stimulations to clearly observe which Vβs were enriched over time and therefore considered to be specific to epitope recognition. [Figure 14-2] Cellular fluorescence immunoprofiling of T cell populations. Characterization of the Vβ repertoire. (A) Clonal proliferation of cyclin A1-specific T cells isolated from HD1, HD2, and HD3 was evaluated by flow cytometry. For HD1 T cells, analysis was performed after 5 and 7 stimulations to clearly observe which Vβs were enriched over time and therefore considered to be specific to epitope recognition. [Figure 14-3] Cellular fluorescence immunoprofiling of T cell populations. Characterization of the Vβ repertoire. For HD2(B) and HD3(C) T cells, the inventors observed a clear expansion of one Vβ chain in HD2 and two Vβ chains in HD3. TRBV, T cell receptor variable region of the β chain. [Figure 15-1]Longitudinal TCR sequencing of enriched cyclin A1-specific T cells. T cells generated from three healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in a cyclin A1 pool. Sequencing results showed the presence of dominant clonal types for different passages derived from HD1(A-E). The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated, in gray shading. The remaining sequences are grouped together in white. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 15-2] Longitudinal TCR sequencing of enriched cyclin A1-specific T cells. T cells generated from three healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in a cyclin A1 pool. Sequencing results showed the presence of dominant clonal types for different passages derived from HD1(A-E). The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated, in gray shading. The remaining sequences are grouped together in white. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 15-3]Longitudinal TCR sequencing of enriched cyclin A1-specific T cells. T cells generated from three healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in a cyclin A1 pool. Sequencing results showed the presence of dominant clonal types for different passages derived from HD1(A-E). The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated, in gray shading. The remaining sequences are grouped together in white. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 15-4] Longitudinal TCR sequencing of enriched cyclin A1-specific T cells. T cells generated from three healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in a cyclin A1 pool. Sequencing results showed the presence of dominant clonal types for different passages derived from HD1(A-E). The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated, in gray shading. The remaining sequences are grouped together in white. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 15-5]Longitudinal TCR sequencing of enriched cyclin A1-specific T cells. T cells generated from three healthy donors included in the experimental setup were characterized by TCRαβ sequencing after several stimulations in a cyclin A1 pool. Sequencing results showed the presence of dominant clonal types for different passaged cultures derived from HD2(F) and HD3(G). The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated, in gray shading. The remaining sequences are grouped together in white. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 16] Genesis and functional verification of cyclin A1-specific T cells using TCR gene transfection and TCR gene editing approaches. (A) Disruption of the endogenous TCR repertoire and expression of different isolated TCRs in edited T cells were evaluated by measuring the surface expression of CD3 and specific Vβ in untransduced T cells or T cells transduced with LV, respectively (B, C). (D) Functionality of transgenic T cells was evaluated by measuring CD107a expression and IFNγ secretion in CD8+ and CD4+ T cells during 6-hour co-culture with autotarget cells pulsed with specific peptides or unrelated peptides. Either transferred cells or TCR-edited cells were used as effector cells in the assay. P: peptide, SP: subpool, UT: untransduced, ED: edited, TR: transferred. [Figure 17]HLA restriction of cyclin A1-specific T lymphocytes. (A) Table showing EBV cell lines used to identify HLA restriction in cyclin A1-HD1 T cell passages specific to P102 and P52 / P53. HLA elements present in EBV cells and shared with HD1 are highlighted. In EBV-BLCL#9 (left) pulsed with P52 / P53, IFNγ production and CD107a secretion were observed, indicating that the restricting element of the P52 / 53 TCR is HLA-B*35:02. In EBV-BLCL#12 (right) pulsed with P102, the restricting element of the P102 TCR was indicated to be either HLA-DRB1*07:01 or HLA-DQB1*02:02. (B) The inventors confirmed the results by performing a functional assay using only the EBV cell line recognized in the first assay as the target cell and transgenic T cells generated using either a TCR transfer or a TCR gene editing protocol as the effector cell. Both TCRs were able to activate CD4 and CD8 T cells. HLA: Human leukocyte antigen, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, P: Peptide, unr: Unrelated peptide, HD: Healthy donor. [Figure 18] Elimination of primary leukemia blasts by T cells engineered to express a TCR specific to the P52 / P53 peptide nonamer. Evaluation of target cell death by cell fluorescence measurement, measured by determining caspase-3 activation during co-culture with edited T cells. AML1-derived blasts express the HLA-DRB1*07:01 and HLA-DQB1*02:02 alleles, while AML2 blasts express the HLA-B*35:02 allele. ED: Edited, P: Peptide, AML: Acute myeloid leukemia. [Figure 19] The proliferation of tumor-specific T cells from peripheral blood mononuclear cells, and the frequencies of CD107a, IFN-γ, TNF-α, and IL-2-positive cells, are shown after a subsequent stimulation round in T cells co-cultured for 6 hours with (A) PDAC or (B) CRC-specific peptide pools, or with autologous APCs loaded with unrelated peptides. [Figure 20-1]Identification of tumor immunogenic peptides: The frequencies of CD107a, IFN-γ, TNF-α, and IL-2-positive cells are shown for PDAC (A-C, left panel) in T cells co-cultured for 6 hours with autologous APCs loaded with individual peptide subpools. The mapping grid defines the peptide subpools, with each peptide present in only two crossing subpools. Peptide-specific recognition was confirmed during co-culture with peptide-pulsed autologous APCs or T2 cell lines (A-E, right panel). [Figure 20-2] Identification of tumor immunogenic peptides: The frequencies of CD107a, IFN-γ, TNF-α, and IL-2-positive cells are shown for PDAC (A-C, left panel) in T cells co-cultured for 6 hours with autologous APCs loaded with individual peptide subpools. The mapping grid defines the peptide subpools, with each peptide present in only two crossing subpools. Peptide-specific recognition was confirmed during co-culture with peptide-pulsed autologous APCs or T2 cell lines (A-E, right panel). [Figure 20-3] Identification of tumor immunogenic peptides: The frequencies of CD107a, IFN-γ, TNF-α, and IL-2-positive cells are shown for PDAC (A-C, left panel) in T cells co-cultured for 6 hours with autologous APCs loaded with individual peptide subpools. The mapping grid defines the peptide subpools, with each peptide present in only two crossing subpools. Peptide-specific recognition was confirmed during co-culture with peptide-pulsed autologous APCs or T2 cell lines (A-E, right panel). [Figure 20-4] Identification of tumor immunogenic peptides: The frequencies of CD107a, IFN-γ, TNF-α, and IL-2-positive cells are shown for CRCs (D, E, left panel) in T cells co-cultured for 6 hours with autologous APCs loaded with individual peptide subpools. The mapping grid defines the peptide subpools, with each peptide present in only two crossing subpools. Peptide-specific recognition was confirmed during co-culture with peptide-pulsed autologous APCs or T2 cell lines (A-E, right panel). [Figure 20-5]Identification of tumor immunogenic peptides: The frequencies of CD107a, IFN-γ, TNF-α, and IL-2-positive cells are shown for CRCs (D, E, left panel) in T cells co-cultured for 6 hours with autologous APCs loaded with individual peptide subpools. The mapping grid defines the peptide subpools, with each peptide present in only two crossing subpools. Peptide-specific recognition was confirmed during co-culture with peptide-pulsed autologous APCs or T2 cell lines (A-E, right panel). [Figure 21-1] Clone tracking of tumor-specific T cells. Generated T cells were characterized by TCRαβ sequencing after several stimulations in PDAC or CRC pools. Sequencing results showed the presence of a dominant clonal type for all T cell cultures analyzed. The pie charts show the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. (A) DKK1 TCR, (B) MUC5AC TCR. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 21-2]Clone tracking of tumor-specific T cells. Generated T cells were characterized by TCRαβ sequencing after several stimulations in a PDAC or CRC pool. Sequencing results showed the presence of a dominant clonal type for all T cell cultures analyzed. The pie charts show the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. (C) AGR2 TCR, (D) c-MET TCR, (E) MSLN TCR. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linkage gene, TRBJ: T cell receptor β chain linkage gene, TRBD: T cell receptor β chain diversity gene. [Figure 21-3] Clone tracking of tumor-specific T cells. Generated T cells were characterized by TCRαβ sequencing after several stimulations in a PDAC or CRC pool. Sequencing results showed the presence of a dominant clonal type for all T cell cultures analyzed. The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. (F)HER2 TCR. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linking gene, TRBJ: T cell receptor β chain linking gene, TRBD: T cell receptor β chain diversity gene. [Figure 21-4]Clone tracking of tumor-specific T cells. Generated T cells were characterized by TCRαβ sequencing after several stimulations in a PDAC or CRC pool. Sequencing results showed the presence of a dominant clonal type for all T cell cultures analyzed. The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. (G)CTDP1 TCR. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linking gene, TRBJ: T cell receptor β chain linking gene, TRBD: T cell receptor β chain diversity gene. [Figure 21-5] Tumor-Specific T Cell Clonal Tracking: Generated T cells were characterized by TCRαβ sequencing after several stimulations in a PDAC or CRC pool. Sequencing results showed the presence of a dominant clonal type for all T cell cultures analyzed. The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. (H)TACC2 TCR. CDR3: Complementarity Determining Region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linking gene, TRBJ: T cell receptor β chain linking gene, TRBD: T cell receptor β chain diversity gene. [Figure 21-6]Clone tracking of tumor-specific T cells. Generated T cells were characterized by TCRαβ sequencing after several stimulations in a PDAC or CRC pool. Sequencing results showed the presence of a dominant clonal type for all T cell cultures analyzed. The pie chart shows the contribution of the most dominant CDR3 amino acid sequence identified at the last time point (e.g., S8 corresponds to the sequencing result obtained after the 8th stimulation) to the TCR repertoire at each stimulation evaluated. The remaining sequences are grouped together in white. (I) PTGFRN TCR. CDR3: Complementarity-determining region 3, S: Stimulus, TRAV: T cell receptor α chain variable gene, TRBV: T cell receptor β chain variable gene, TRAJ: T cell receptor α chain linking gene, TRBJ: T cell receptor β chain linking gene, TRBD: T cell receptor β chain diversity gene. [Figure 22-1]In vitro recognition and elimination of HER2+HLA-A2+ cancer cell lines and patient-derived organoids by proliferated HER2-specific T cells (A) HER2-transgenic T cells (n=3) were co-cultured for 24 hours with HLA-A0201+PDAC cell lines (PANC1, BxPCR-A2+, T3M4) at an effector-to-target ratio of 5:1. Cell death was expressed as the area (μm2) covered by cells undergoing apoptosis (caspase 3 / 7 activation) over time. (B) HER2-transgenic T cells (n=5) were co-cultured for 24 hours with luciferase+HLA-A0201+PDAC cell lines (PANC1, T3M4, and BxPC3-A2+) and HLA-A0201-on (BxPC3-A2-) as a control at an effector-to-target ratio of 1:1. Specific cell death was assessed by measuring luciferase activity 24 hours after co-culture. The exclusion index was calculated as [1 - (luciferase activity of target cell line with T cells / luciferase activity of target cell line alone)]. (C) HER2-transgenic T cells (n=3) were co-cultured for 48 hours with a colorectal cancer organoid (PDO) derived from one patient at an effector-to-target ratio of 5:1. Cell death was expressed as the area (μm2) covered by apoptotic cells (caspase 3 / 7 activated). (D) Timeline of the in vivo experiment. Luciferase-introduced PDO was transplanted into the liver. Three days later, T cells were injected into the liver, and on day 10, a second dose of T cells was delivered intravenously. Bioluminescence assessment of tumor growth (E). Data are expressed as mean ± SD. Statistical significance was assessed by linear regression curves fitted to observational results (A, C) or two-way ANOVA (B, E). ***P<0.001 and ****P<0.0001. [Figure 22-2]In vitro recognition and elimination of HER2+HLA-A2+ cancer cell lines and patient-derived organoids by proliferated HER2-specific T cells (A) HER2-transgenic T cells (n=3) were co-cultured for 24 hours with HLA-A0201+PDAC cell lines (PANC1, BxPCR-A2+, T3M4) at an effector-to-target ratio of 5:1. Cell death was expressed as the area (μm2) covered by cells undergoing apoptosis (caspase 3 / 7 activation) over time. (B) HER2-transgenic T cells (n=5) were co-cultured for 24 hours with luciferase+HLA-A0201+PDAC cell lines (PANC1, T3M4, and BxPC3-A2+) and HLA-A0201-on (BxPC3-A2-) as a control at an effector-to-target ratio of 1:1. Specific cell death was assessed by measuring luciferase activity 24 hours after co-culture. The exclusion index was calculated as [1 - (luciferase activity of target cell line with T cells / luciferase activity of target cell line alone)]. (C) HER2-transgenic T cells (n=3) were co-cultured for 48 hours with a colorectal cancer organoid (PDO) derived from one patient at an effector-to-target ratio of 5:1. Cell death was expressed as the area (μm2) covered by apoptotic cells (caspase 3 / 7 activated). (D) Timeline of the in vivo experiment. Luciferase-introduced PDO was transplanted into the liver. Three days later, T cells were injected into the liver, and on day 10, a second dose of T cells was delivered intravenously. Bioluminescence assessment of tumor growth (E). Data are expressed as mean ± SD. Statistical significance was assessed by linear regression curves fitted to observational results (A, C) or two-way ANOVA (B, E). ***P<0.001 and ****P<0.0001. [Figure 22-3]In vitro recognition and elimination of HER2+HLA-A2+ cancer cell lines and patient-derived organoids by proliferated HER2-specific T cells (A) HER2-transgenic T cells (n=3) were co-cultured for 24 hours with HLA-A0201+PDAC cell lines (PANC1, BxPCR-A2+, T3M4) at an effector-to-target ratio of 5:1. Cell death was expressed as the area (μm2) covered by cells undergoing apoptosis (caspase 3 / 7 activation) over time. (B) HER2-transgenic T cells (n=5) were co-cultured for 24 hours with luciferase+HLA-A0201+PDAC cell lines (PANC1, T3M4, and BxPC3-A2+) and HLA-A0201-on (BxPC3-A2-) as a control at an effector-to-target ratio of 1:1. Specific cell death was assessed by measuring luciferase activity 24 hours after co-culture. The exclusion index was calculated as [1 - (luciferase activity of target cell line with T cells / luciferase activity of target cell line alone)]. (C) HER2-transgenic T cells (n=3) were co-cultured for 48 hours with a colorectal cancer organoid (PDO) derived from one patient at an effector-to-target ratio of 5:1. Cell death was expressed as the area (μm2) covered by apoptotic cells (caspase 3 / 7 activated). (D) Timeline of the in vivo experiment. Luciferase-introduced PDO was transplanted into the liver. Three days later, T cells were injected into the liver, and on day 10, a second dose of T cells was delivered intravenously. Bioluminescence assessment of tumor growth (E). Data are expressed as mean ± SD. Statistical significance was assessed by linear regression curves fitted to observational results (A, C) or two-way ANOVA (B, E). ***P<0.001 and ****P<0.0001. [Figure 23]Recognition and death of Ag+HLA-A0201+ cancer cell lines and patient-derived organoids in vitro by proliferated MSLN-specific T cells. (A) MSLN-transgenic T cells (n=3) were co-cultured with HLA-A0201+ PDAC cell line (PANC1) at an effector-to-target ratio of 5:1 for 24 hours. Death was expressed as the area (μm2) covered by target cells undergoing apoptosis (caspase 3 / 7 activation) over time. (B) MSLN-transgenic T cells (n=3) were co-cultured with HLA-incompatible PDAC cell line (BxPC3) as a control at an effector-to-target ratio of 5:1 for 24 hours. Death was expressed as the area (μm2) covered by cells undergoing apoptosis (caspase 3 / 7 activation) over time. (C) MSLN-transgenic T cells (n=3) were co-cultured for 50 hours with patient-derived colorectal cancer organoids from one primary tumor (left) and one liver metastasis (right) at an effector-to-target ratio of 5:1. Data are normalized at the first useful time and expressed as mean ± SEM. Statistical significance was assessed by linear regression curves fitted to the observed results. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 [Figure 24]Recognition and death of Ag+HLA-A0201+ cancer cell lines and patient-derived organoids in vitro by proliferated DKK1-specific T cells. (A) DKK1-transgenic T cells (n=3) were co-cultured with HLA-A0201+ PDAC cell line (PANC1) at an effector-to-target ratio of 5:1 for 24 hours. Death was expressed as the area (μm2) covered by target cells undergoing apoptosis (caspase 3 / 7 activation) over time. (B) DKK1-transgenic T cells (n=3) were co-cultured with HLA-incompatible PDAC cell line (BxPC3) as a control at an effector-to-target ratio of 5:1 for 24 hours. Death was expressed as the area (μm2) covered by cells undergoing apoptosis (caspase 3 / 7 activation) over time. (C) DKK1-transgenic T cells (n=3) were co-cultured for 50 hours with patient-derived colorectal cancer organoids from one primary tumor (left) and one liver metastasis (right) at an effector-to-target ratio of 5:1. Data from the first useful time point are expressed as mean ± SEM. Statistical significance was assessed by linear regression curves fitted to the observed results. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 [Figure 25]Recognition and death of Ag+HLA-A0201+ cancer cell lines and patient-derived organoids in vitro by proliferated AGR2-specific T cells. (A) AGR2-transgenic T cells (n=3) were co-cultured with HLA-A0201+ PDAC cell line (PANC1) at an effector-to-target ratio of 5:1 for 24 hours. Death was expressed as the area (μm2) covered by target cells undergoing apoptosis (caspase 3 / 7 activation) over time. (B) AGR2-transgenic T cells (n=3) were co-cultured with HLA-incompatible PDAC cell line (BxPC3) as a control at an effector-to-target ratio of 5:1 for 24 hours. Death was expressed as the area (μm2) covered by cells undergoing apoptosis (caspase 3 / 7 activation) over time. (C) AGR2-transgenic T cells (n=3) were co-cultured for 50 hours with patient-derived colorectal cancer organoids from one primary tumor (left) at an effector-to-target ratio of 5:1 and one liver metastasis (right) at an effector-to-target ratio of 10:1. Data from the first useful time point are expressed as mean ± SEM. Statistical significance was assessed by linear regression curves fitted to the observed results. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001. [Figure 26]Cytotoxic activity of PR3-TCR-engineered T cells against HLA-A*02:01+ antigen-expressing leukemia cells. (A) Transduction efficiency measured as the percentage of cells expressing TCR expression (left) and CD3 as a surrogate marker for memory phenotype on the surface of edited lymphocytes. (B) Degranulation ability and cytokine secretion in engineered T cells when co-cultured with HLA-A*02:01+ peptide-pulsed T2 cells. (C) Bar graphs show the death efficiency of engineered T cells when co-cultured with leukemia cell lines (K562 and THP1), measured by determining the exclusion index calculated as follows: [1-(target cells in co-culture / target cells in monoculture)]. Graphed data represent the mean ± SEM of 6 biological replicates for the K562 Tg dataset and the mean ± SEM of 5 biological replicates for the THP1 dataset. (D) Cell death assays performed on TCR-edited T cells cultured for 24 hours with HLA-A*02:01+ blast cells (pAML 8 and 9) or pAML 10 (HLA-A*02:01-) as a control. In particular, increased exclusion indices were observed in pAML 9 compared to HLA-incompatible pAML 10 at both 1:1 and 1:5 ratios. Data are presented as bar graphs showing the mean ± SEM of three biological replicates. Memory subsets are defined as follows: TSCM (CD45RA+CD62L+CD95+), TCM (CD45RA-CD62L+), TEM (CD45RA-CD62L-), TEMRA (CD45+CD62L-). PR3: Proteinase 3, HLA: Human leukocyte antigen, unrel: Unrelated, CM: Central memory, SCM: Stem cell memory, EM: Effector memory, EMRA: Terminal differentiation effector memory, E:T: Effector vs. target ratio, pAML: Primary acute myeloid leukemia cells, Interleukin-2 (IL-2), Tumor necrosis factor α (TNF-α), and Interferon-γ (IFN-γ). [Figure 27-1]CD8 coreceptor-dependent target cell recognition by CTSG-TCR transduced T cells. A) Co-culture of engineered T cells with peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+ cells demonstrates that CD8+ TCR-expressing lymphocytes can efficiently recognize target cells (demonstrated with respect to both degranulation and T cell activation state). B) Experimental setup used to improve cell activity by exogenously adding CD8 coreceptors. Briefly, naked T cells are transduced first with a lentiviral vector (LV) encoding the CTSG-TCR chain, and then co-transduced with a lentiviral vector (LV) encoding the CD8αβ chain, upon disruption of the endogenous TCR repertoire. C) Upon co-transduction with CD8-encoding LV, bitransduced CD4+CD8+CTSG-TCR-expressing T cells were able to efficiently recognize peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+EBV-BLCL compared to CD4+TCR-expressing T lymphocytes. Data are presented as bar graphs showing the mean ± SEM of three biological replicates. CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, unrel: Unrelated, EBV-BLCL: Epstein-Barr virus-transformed B lymphoblast-like cell line. [Figure 27-2]CD8 coreceptor-dependent target cell recognition by CTSG-TCR transduced T cells. A) Co-culture of engineered T cells with peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+ cells demonstrates that CD8+ TCR-expressing lymphocytes can efficiently recognize target cells (demonstrated with respect to both degranulation and T cell activation state). B) Experimental setup used to improve cell activity by exogenously adding CD8 coreceptors. Briefly, naked T cells are transduced first with a lentiviral vector (LV) encoding the CTSG-TCR chain, and then co-transduced with a lentiviral vector (LV) encoding the CD8αβ chain, upon disruption of the endogenous TCR repertoire. C) Upon co-transduction with CD8-encoding LV, bitransduced CD4+CD8+CTSG-TCR-expressing T cells were able to efficiently recognize peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+EBV-BLCL compared to CD4+TCR-expressing T lymphocytes. Data are presented as bar graphs showing the mean ± SEM of three biological replicates. CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, unrel: Unrelated, EBV-BLCL: Epstein-Barr virus-transformed B lymphoblast-like cell line. [Figure 27-3]CD8 coreceptor-dependent target cell recognition by CTSG-TCR transduced T cells. A) Co-culture of engineered T cells with peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+ cells demonstrates that CD8+ TCR-expressing lymphocytes can efficiently recognize target cells (demonstrated with respect to both degranulation and T cell activation state). B) Experimental setup used to improve cell activity by exogenously adding CD8 coreceptors. Briefly, naked T cells are transduced first with a lentiviral vector (LV) encoding the CTSG-TCR chain, and then co-transduced with a lentiviral vector (LV) encoding the CD8αβ chain, upon disruption of the endogenous TCR repertoire. C) Upon co-transduction with CD8-encoding LV, bitransduced CD4+CD8+CTSG-TCR-expressing T cells were able to efficiently recognize peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+EBV-BLCL compared to CD4+TCR-expressing T lymphocytes. Data are presented as bar graphs showing the mean ± SEM of three biological replicates. CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, unrel: Unrelated, EBV-BLCL: Epstein-Barr virus-transformed B lymphoblast-like cell line. [Figure 28-1]Production of CTSG-TCR T cells. A) Experimental layout used to manipulate T cells expressing CTSG-TCR alone (CTSG-TCR T cells) or T cells expressing CTSG-TCR in combination with the CD8 coreceptor (CTSG-TCR+CD8 T cells). T cells were grown using a G-Rex cell culture plate during TCR gene editing. B) Editing efficiency (left panel), memory phenotype (center panel), and CD4 / CD8 subset composition (right panel) of the manipulated T cell product. C) Evaluation of in vitro T cell specificity when co-cultured with peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+ EBV-BLCL. The results obtained with G-Rex proliferating T cells replicated the ability of lymphocytes to recognize target cells in an HLA-specific manner, and the increased ability of bitransduced CD4+CD8+TCR+ T cells (CTSG-TCR CD4+CD8+ T cells) to degranulate (upper panel) and be activated (lower panel) compared to monotransduced CD4+TCR+ lymphocytes (CTSG-TCR CD4+ T cells). Data are presented as bar graphs showing the mean ± SEM of three biological replicates. Memory subsets are defined as follows: TSCM (CD45RA+CD62L+CD95+), TCM (CD45RA-CD62L+), TEM (CD45RA-CD62L-), and TEMRA (CD45+CD62L-). CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, unrel: Unrelated, CM: Central memory, SCM: Stem cell memory, EM: Effector memory, EMRA: Terminal differentiation effector memory, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line. [Figure 28-2]Production of CTSG-TCR T cells. A) Experimental layout used to manipulate T cells expressing CTSG-TCR alone (CTSG-TCR T cells) or T cells expressing CTSG-TCR in combination with the CD8 coreceptor (CTSG-TCR+CD8 T cells). T cells were grown using a G-Rex cell culture plate during TCR gene editing. B) Editing efficiency (left panel), memory phenotype (center panel), and CD4 / CD8 subset composition (right panel) of the manipulated T cell product. C) Evaluation of in vitro T cell specificity when co-cultured with peptide-pulsed HLA-A*24:02+ or HLA-C*07:02+ EBV-BLCL. The results obtained with G-Rex proliferating T cells replicated the ability of lymphocytes to recognize target cells in an HLA-specific manner, and the increased ability of bitransduced CD4+CD8+TCR+ T cells (CTSG-TCR CD4+CD8+ T cells) to degranulate (upper panel) and be activated (lower panel) compared to monotransduced CD4+TCR+ lymphocytes (CTSG-TCR CD4+ T cells). Data are presented as bar graphs showing the mean ± SEM of three biological replicates. Memory subsets are defined as follows: TSCM (CD45RA+CD62L+CD95+), TCM (CD45RA-CD62L+), TEM (CD45RA-CD62L-), and TEMRA (CD45+CD62L-). CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, unrel: Unrelated, CM: Central memory, SCM: Stem cell memory, EM: Effector memory, EMRA: Terminal differentiation effector memory, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line. [Figure 29-1]Sustained proliferation and proliferation of manipulated T cells in leukemia-bearing mice. A) Study design for an in vivo trial. Timeline of the AML in vivo model. On day 3, mice were intravenously injected with 1.5–2 × 10⁶ pAML blast cells, followed by injection of 10 × 10⁶ CTSG-TCR-expressing edited T cells on day 0. Longitudinal monitoring of T cell and leukemia dynamics was evaluated weekly by posterior orbital blood sampling. The schematic diagram shows two different mouse groups based on the injected leukemia. Group 1 (upper graph) was injected with HLA-A*24:02+ blast cells, where n=6 mice were treated with CTSG-TCR+CD8 transduced T cells and 4 mice were left untreated. Group 2 (lower graph) was injected with HLA-C*07:02+ leukemic blast cells. In this group, nine mice were left untreated, while n=11 and n=7 were injected with CTSG-TCR-specific T cells co-transduced or not co-transduced with a CD8 construct, respectively. At sacrifice, the tongue, ear, spleen, bone marrow, and peripheral blood were collected. B) Persistence of TCR-T cells in the peripheral circulation of treated mice. HLA-A*24:02+AML-carrying mice injected with CTSG TCR+CD8 T cells (left panel, graphed data are represented as the mean ± SEM of 6 biological replicas), and HLA-C*07:02+AML-carrying mice injected with CTSG TCR+CD8 T cells or CTSG TCR alone (right panel, graphed data are represented as the mean ± SEM of 11 biological replicas for the CTSG+CD8 subgroup and 7 biological replicas for the CTSG subgroup). CD4 / CD8 subset composition (C) and memory phenotype (D) of engineered T cell products used to treat mice injected with HLA-A*24:02+ (left panel) and HLA-C*07:02+ (right panel: upper graph represents CTSG-TCR+CD8 T cells, lower graph represents CTSG-TCR T cells). Memory subsets are defined as follows: TSCM (CD45RA+CD62L+CD95+), TCM (CD45RA-CD62L+), TEM (CD45RA-CD62L-), TEMRA (CD45+CD62L-). CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, pAML: Primary acute myeloid leukemia cell. [Figure 29-2]Sustained proliferation and proliferation of manipulated T cells in leukemia-bearing mice. A) Study design for an in vivo trial. Timeline of the AML in vivo model. On day 3, mice were intravenously injected with 1.5–2 × 10⁶ pAML blast cells, followed by injection of 10 × 10⁶ CTSG-TCR-expressing edited T cells on day 0. Longitudinal monitoring of T cell and leukemia dynamics was evaluated weekly by posterior orbital blood sampling. The schematic diagram shows two different mouse groups based on the injected leukemia. Group 1 (upper graph) was injected with HLA-A*24:02+ blast cells, where n=6 mice were treated with CTSG-TCR+CD8 transduced T cells and 4 mice were left untreated. Group 2 (lower graph) was injected with HLA-C*07:02+ leukemic blast cells. In this group, nine mice were left untreated, while n=11 and n=7 were injected with CTSG-TCR-specific T cells co-transduced or not co-transduced with a CD8 construct, respectively. At sacrifice, the tongue, ear, spleen, bone marrow, and peripheral blood were collected. B) Persistence of TCR-T cells in the peripheral circulation of treated mice. HLA-A*24:02+AML-carrying mice injected with CTSG TCR+CD8 T cells (left panel, graphed data are represented as the mean ± SEM of 6 biological replicas), and HLA-C*07:02+AML-carrying mice injected with CTSG TCR+CD8 T cells or CTSG TCR alone (right panel, graphed data are represented as the mean ± SEM of 11 biological replicas for the CTSG+CD8 subgroup and 7 biological replicas for the CTSG subgroup). CD4 / CD8 subset composition (C) and memory phenotype (D) of engineered T cell products used to treat mice injected with HLA-A*24:02+ (left panel) and HLA-C*07:02+ (right panel: upper graph represents CTSG-TCR+CD8 T cells, lower graph represents CTSG-TCR T cells). Memory subsets are defined as follows: TSCM (CD45RA+CD62L+CD95+), TCM (CD45RA-CD62L+), TEM (CD45RA-CD62L-), TEMRA (CD45+CD62L-). CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, pAML: Primary acute myeloid leukemia cell. [Figure 29-3]Sustained proliferation and proliferation of manipulated T cells in leukemia-bearing mice. A) Study design for an in vivo trial. Timeline of the AML in vivo model. On day 3, mice were intravenously injected with 1.5–2 × 10⁶ pAML blast cells, followed by injection of 10 × 10⁶ CTSG-TCR-expressing edited T cells on day 0. Longitudinal monitoring of T cell and leukemia dynamics was evaluated weekly by posterior orbital blood sampling. The schematic diagram shows two different mouse groups based on the injected leukemia. Group 1 (upper graph) was injected with HLA-A*24:02+ blast cells, where n=6 mice were treated with CTSG-TCR+CD8 transduced T cells and 4 mice were left untreated. Group 2 (lower graph) was injected with HLA-C*07:02+ leukemic blast cells. In this group, nine mice were left untreated, while n=11 and n=7 were injected with CTSG-TCR-specific T cells co-transduced or not co-transduced with a CD8 construct, respectively. At sacrifice, the tongue, ear, spleen, bone marrow, and peripheral blood were collected. B) Persistence of TCR-T cells in the peripheral circulation of treated mice. HLA-A*24:02+AML-carrying mice injected with CTSG TCR+CD8 T cells (left panel, graphed data are represented as the mean ± SEM of 6 biological replicas), and HLA-C*07:02+AML-carrying mice injected with CTSG TCR+CD8 T cells or CTSG TCR alone (right panel, graphed data are represented as the mean ± SEM of 11 biological replicas for the CTSG+CD8 subgroup and 7 biological replicas for the CTSG subgroup). CD4 / CD8 subset composition (C) and memory phenotype (D) of engineered T cell products used to treat mice injected with HLA-A*24:02+ (left panel) and HLA-C*07:02+ (right panel: upper graph represents CTSG-TCR+CD8 T cells, lower graph represents CTSG-TCR T cells). Memory subsets are defined as follows: TSCM (CD45RA+CD62L+CD95+), TCM (CD45RA-CD62L+), TEM (CD45RA-CD62L-), TEMRA (CD45+CD62L-). CTSG or CG: Cathepsin G, HLA: Human leukocyte antigen, pAML: Primary acute myeloid leukemia cell. [Figure 30-1]In vivo evaluation of T-cell antitumor effects. A) AML proliferation during the experiment was measured as primary leukemia blast count / μL. The left panel shows the leukemia burden in untreated or HLA-A*24:02+AML-carrying mice injected with CTSG TCR+CD8 T cells (graphed data are represented as the mean ± SEM of 6 biological replicates for the treatment group and 4 for the control group). The right panel shows the leukemia burden in untreated or HLA-C*07:02+AML-carrying mice injected with CTSG TCR+CD8 T cells and CTSG TCR T cells (data are represented as the mean ± SEM of 7 biological replicates for the CTSG subgroup, 11 biological replicates for the CTSG+CD8 subgroup, and 9 biological replicates for the control group). B) Graphs showing the percentage of blast cells present in bone marrow and spleen samples at sacrifice. The results demonstrated the efficacy of CTSG-TCR T cells and CTSG-TCR+CD8-modified T cells in reducing tumor growth. Conversely, tumor enlargement was observed in untreated mice for both leukemias. Data are presented as mean ± SEM from six biological replicates for the CTSG+CD8 subgroup, four biological replicates for the HLA-A*24:02+pAML control group (left graph), seven biological replicates for the CTSG subgroup, eleven biological replicates for the CTSG+CD8 subgroup, and five biological replicates for the HLA-C*07:02+pAML control group (right graph). C) The graph shows the time course of CD3+ dynamics in HLA-C*07:02+-carrying mice treated with CTSG+CD8-modified T cells. These results highlight poor T cell engraftment over time in mice 17 and 18 compared to the rest of the mice (CTSG+CD8 dataset), and therefore likely explain the reduced control of leukemia burden in mouse bone marrow. Indeed, mice 17 and 18 represent two mice in which CTSG+CD8 edited T cell treatment appears not to have efficiently controlled leukemia proliferation (as indicated by triangles and squares in the blast % in bone marrow in the central graph B). Data for the CTSG+CD8 subgroup are presented as mean ± SEM of nine biological replicates.D) The graphs show the weight changes of each mouse at various time points during the in vivo study, calculated as [(body weight at each time point / baseline body weight before T cell injection) × 100] (each line represents a mouse). The left panel shows the results for mice injected with untreated HLA-A*24:02+ leukemic blasts (top graph) and n=6 mice treated with CTSG TCR+CD8 T cells (bottom graph). The right panel shows the weight trends of mice with HLA-C*07:02+: n=9 untreated mice (top graph), n=11 mice injected with CTSG TCR+CD8 T cells (center graph), and n=7 mice injected with CTSG TCR T cells (bottom graph). Statistical significance was evaluated by a two-way ANOVA multiple comparison test, and the resulting p-values ​​are recorded as ****p<0.0001. CTSG: Cathepsin G, HLA: Human Leukocyte Antigen. [Figure 30-2]In vivo evaluation of T-cell antitumor effects. A) AML proliferation during the experiment was measured as primary leukemia blast count / μL. The left panel shows the leukemia burden in untreated or HLA-A*24:02+AML-carrying mice injected with CTSG TCR+CD8 T cells (graphed data are represented as the mean ± SEM of 6 biological replicates for the treatment group and 4 for the control group). The right panel shows the leukemia burden in untreated or HLA-C*07:02+AML-carrying mice injected with CTSG TCR+CD8 T cells and CTSG TCR T cells (data are represented as the mean ± SEM of 7 biological replicates for the CTSG subgroup, 11 biological replicates for the CTSG+CD8 subgroup, and 9 biological replicates for the control group). B) Graphs showing the percentage of blast cells present in bone marrow and spleen samples at sacrifice. The results demonstrated the efficacy of CTSG-TCR T cells and CTSG-TCR+CD8-modified T cells in reducing tumor growth. Conversely, tumor enlargement was observed in untreated mice for both leukemias. Data are presented as mean ± SEM from six biological replicates for the CTSG+CD8 subgroup, four biological replicates for the HLA-A*24:02+pAML control group (left graph), seven biological replicates for the CTSG subgroup, eleven biological replicates for the CTSG+CD8 subgroup, and five biological replicates for the HLA-C*07:02+pAML control group (right graph). C) The graph shows the time course of CD3+ dynamics in HLA-C*07:02+-carrying mice treated with CTSG+CD8-modified T cells. These results highlight poor T cell engraftment over time in mice 17 and 18 compared to the rest of the mice (CTSG+CD8 dataset), and therefore likely explain the reduced control of leukemia burden in mouse bone marrow. Indeed, mice 17 and 18 represent two mice in which CTSG+CD8 edited T cell treatment appears not to have efficiently controlled leukemia proliferation (as indicated by triangles and squares in the blast % in bone marrow in the central graph B). Data for the CTSG+CD8 subgroup are presented as mean ± SEM of nine biological replicates.D) The graphs show the weight changes of each mouse at various time points during the in vivo study, calculated as [(body weight at each time point / baseline body weight before T cell injection) × 100] (each line represents a mouse). The left panel shows the results for mice injected with untreated HLA-A*24:02+ leukemic blasts (top graph) and n=6 mice treated with CTSG TCR+CD8 T cells (bottom graph). The right panel shows the weight trends of mice with HLA-C*07:02+: n=9 untreated mice (top graph), n=11 mice injected with CTSG TCR+CD8 T cells (center graph), and n=7 mice injected with CTSG TCR T cells (bottom graph). Statistical significance was evaluated by a two-way ANOVA multiple comparison test, and the resulting p-values ​​are recorded as ****p<0.0001. CTSG: Cathepsin G, HLA: Human Leukocyte Antigen. [Figure 30-3]In vivo evaluation of T-cell antitumor effects. A) AML proliferation during the experiment was measured as primary leukemia blast count / μL. The left panel shows the leukemia burden in untreated or HLA-A*24:02+AML-carrying mice injected with CTSG TCR+CD8 T cells (graphed data are represented as the mean ± SEM of 6 biological replicates for the treatment group and 4 for the control group). The right panel shows the leukemia burden in untreated or HLA-C*07:02+AML-carrying mice injected with CTSG TCR+CD8 T cells and CTSG TCR T cells (data are represented as the mean ± SEM of 7 biological replicates for the CTSG subgroup, 11 biological replicates for the CTSG+CD8 subgroup, and 9 biological replicates for the control group). B) Graphs showing the percentage of blast cells present in bone marrow and spleen samples at sacrifice. The results demonstrated the efficacy of CTSG-TCR T cells and CTSG-TCR+CD8-modified T cells in reducing tumor growth. Conversely, tumor enlargement was observed in untreated mice for both leukemias. Data are presented as mean ± SEM from six biological replicates for the CTSG+CD8 subgroup, four biological replicates for the HLA-A*24:02+pAML control group (left graph), seven biological replicates for the CTSG subgroup, eleven biological replicates for the CTSG+CD8 subgroup, and five biological replicates for the HLA-C*07:02+pAML control group (right graph). C) The graph shows the time course of CD3+ dynamics in HLA-C*07:02+-carrying mice treated with CTSG+CD8-modified T cells. These results highlight poor T cell engraftment over time in mice 17 and 18 compared to the rest of the mice (CTSG+CD8 dataset), and therefore likely explain the reduced control of leukemia burden in mouse bone marrow. Indeed, mice 17 and 18 represent two mice in which CTSG+CD8 edited T cell treatment appears not to have efficiently controlled leukemia proliferation (as indicated by triangles and squares in the blast % in bone marrow in the central graph B). Data for the CTSG+CD8 subgroup are presented as mean ± SEM of nine biological replicates.D) The graphs show the weight changes of each mouse at various time points during the in vivo study, calculated as [(body weight at each time point / baseline body weight before T cell injection) × 100] (each line represents a mouse). The left panel shows the results for mice injected with untreated HLA-A*24:02+ leukemic blasts (top graph) and n=6 mice treated with CTSG TCR+CD8 T cells (bottom graph). The right panel shows the weight trends of mice with HLA-C*07:02+: n=9 untreated mice (top graph), n=11 mice injected with CTSG TCR+CD8 T cells (center graph), and n=7 mice injected with CTSG TCR T cells (bottom graph). Statistical significance was evaluated by a two-way ANOVA multiple comparison test, and the resulting p-values ​​are recorded as ****p<0.0001. CTSG: Cathepsin G, HLA: Human Leukocyte Antigen. [Figure 31] Maintenance of specificity and functionality in ex vivo recovered TCR T cells. Lymphocytes collected from the spleens of selected mice treated with engineered T cell lines or CTSG-TCR+CD8 T cell lines, infused with HLA-A*24:02+ or HLA-C*07:02+ pAMLs, were tested in vitro for their function by measuring their degranulation ability (left panel) and their activation status (right panel). The number of biological replicates is reported in the figure. CTSG: Cathepsin G, HLA: Human leukocyte antigen, EBV-BLCL: Epstein-Barr virus transformed B lymphoblastoid cell line, pAML: Primary acute myeloid leukemia cells. [Figure 32]Phenotypes of manipulated cyclin A1-specific TCR T cells. A) Bar graph showing CD3 surface expression (%) in a population of living T cells used as a substitute for TCR complex presentation. The results showed near-complete knockout efficiency, as measured in edited LV non-transduced (CTR) T cells, and high transduction efficiency, as measured in LV transduced T cells. T cells edited for each cyclin A1 epitope are named TCRpYAE, TCRSP18, and TCRp102. Data are expressed as mean ± SEM of six biological replicates. B) Bar graph showing memory subsets in CD3+ edited T cells: TSCM (CD45RA+CD62L+CD95+), TCM (CD45RA-CD62L+), TEM (CD45RA-CD62L-), TEMRA (CD45+CD62L-). Data are expressed as mean ± SEM of six biological replicates. [Figure 33]Recognition of SP18-specific TCR-engineered T cells for the P75 epitope presented on the HLA-C*04:01 allele. A) Bar graph showing CD107a expression in CD3+ T cells after co-culture of T cells from a donor from which tumor-specific T cell clones were isolated with SP18-pulsed (associated) EBV-BLCL (represented by different fill patterns) sharing at least one HLA constraint. EBV-BLCL pulsed with an unrelated peptide was included as a negative control. Data show the mean ± SEM of three biological replicate experiments. p-values ​​were determined using a two-way ANOVA multiple comparison test: *p<0.05. B) Bar graph shows intracellular accumulation of cytokines associated with CD8+ T cell activation: interleukin-2 (IL-2), tumor necrosis factor α (TNF-α), and interferon-γ (IFN-γ). T cells were exposed to HLA-A*33:01 and HLA-C*04:01 EBV-BLCL pulsed with either subpool 18 (SP18) or an unrelated peptide. Data represent the mean ± SEM of three biological replicates. C-D) Bar graphs show the expression of the degranulation marker CD107a (C) and the secretion of pro-inflammatory cytokines IL-2, TNF-α, and IFN-γ (D) by CD8+ T lymphocytes after 6 hours of co-culture with HLA-C*04:01+EBV pulsed with each peptide in SP18 or an unrelated control peptide. Data represent the mean ± SEM of three biological replicates. E) Bar graphs show the killing efficiency of manipulated T cells, measured by determining an exclusion index calculated as follows: [1-(target cells in co-culture / target cells in single culture)]. T cells co-cultured with HLA-C*04:01+ EBV cells pulsed with p75 showed increased killing efficacy compared to those encountered with EBV cells pulsed with unrelated peptides. Graphed data represent the mean ± SEM of three biological replicate experiments. EBV-BLCL, Epstein-Barr virus-transformed B lymphoblastoid cell line. [Figure 34-1]Recognition of HLA class I and HLA class II restricted epitopes by TCRp53 and TCRp102-engineered T cells, respectively. A) Bar graphs showing CD107a expression (degranulation marker) in T cells engineered with two different TCRs redirected to cyclin A1 pYAE or p102 epitopes, pulsed with immunogenic peptides (related peptides) and stimulated with HLA-B*35:02+ and HLA-DRB1*07:04+HLA-DQB1*02:02+EBV-BLCL, respectively. As a negative control, EBV-BLCL was also pulsed with an unrelated peptide. The left and right panels represent the percentage of degranulation marker CD107a evaluated in CD3+CD8+ or CD3+CD4+ viable T cells, respectively. Data are presented as mean ± SEM of six biological replicates. B-C): (B) Bar graphs representing cytokines released intracellularly in TCR pYAE-edited T cells co-cultured with HLA-B*35:02+EBV-BLCL pulsed with pYAE, and (C) TCR p102 T cells co-cultured with EBV-BLCL pulsed with HLA-DRB1*07:04+HLA-DQB1*02:02+p102. Each EBV cell line was pulsed with an unrelated peptide as a negative control. Data are expressed as the mean ± SEM of six biological replicates. D) TCR-T cells targeting the cyclin A1 tumor antigen-derived p102 epitope (right graph) and pYAE epitope (left graph) were analyzed for their antitumor effects. T cell killing activity was determined by measuring the exclusion index after 24 hours of co-culture with HLA-restricted EBV-BLCL pulsed with either the related peptide or the unrelated peptide (negative control). The presented graph displays data as the mean ± SEM of six biological replicates. E) In the context of the cell death assay, T cells redirected to the tumor expressed the activation marker CD137, thereby confirming TCR activation upon recognition of the corresponding peptide. Statistical significance was assessed by a two-way ANOVA multiple comparison test, and the resulting p-values ​​are recorded as *p<0.05, **p<0.01, ****p<0.0001.EBV-BLCLs: Epstein-Barr virus transformed B lymphoblastoid cell lines, interleukin-2 (IL-2): IL-2, tumor necrosis factor α (TNF-α): TNF-α, and interferon-γ (IFN-γ): IFN-γ. [Figure 34-2]Recognition of HLA class I and HLA class II restricted epitopes by TCRp53 and TCRp102-engineered T cells, respectively. A) Bar graphs showing CD107a expression (degranulation marker) in T cells engineered with two different TCRs redirected to cyclin A1 pYAE or p102 epitopes, pulsed with immunogenic peptides (related peptides) and stimulated with HLA-B*35:02+ and HLA-DRB1*07:04+HLA-DQB1*02:02+EBV-BLCL, respectively. As a negative control, EBV-BLCL was also pulsed with an unrelated peptide. The left and right panels represent the percentage of degranulation marker CD107a evaluated in CD3+CD8+ or CD3+CD4+ viable T cells, respectively. Data are presented as mean ± SEM of six biological replicates. B-C): (B) Bar graphs representing cytokines released intracellularly in TCR pYAE-edited T cells co-cultured with HLA-B*35:02+EBV-BLCL pulsed with pYAE, and (C) TCR p102 T cells co-cultured with EBV-BLCL pulsed with HLA-DRB1*07:04+HLA-DQB1*02:02+p102. Each EBV cell line was pulsed with an unrelated peptide as a negative control. Data are expressed as the mean ± SEM of six biological replicates. D) TCR-T cells targeting the cyclin A1 tumor antigen-derived p102 epitope (right graph) and pYAE epitope (left graph) were analyzed for their antitumor effects. T cell killing activity was determined by measuring the exclusion index after 24 hours of co-culture with HLA-restricted EBV-BLCL pulsed with either the related peptide or the unrelated peptide (negative control). The presented graph displays data as the mean ± SEM of six biological replicates. E) In the context of the cell death assay, T cells redirected to the tumor expressed the activation marker CD137, thereby confirming TCR activation upon recognition of the corresponding peptide. Statistical significance was assessed by a two-way ANOVA multiple comparison test, and the resulting p-values ​​are recorded as *p<0.05, **p<0.01, ****p<0.0001.EBV-BLCLs: Epstein-Barr virus transformed B lymphoblastoid cell lines, interleukin-2 (IL-2): IL-2, tumor necrosis factor α (TNF-α): TNF-α, and interferon-γ (IFN-γ): IFN-γ. [Figure 35] sgRNA scheme. A scheme of sgRNAs designed to target exon 3 of TRAC, exon 1 of TRBC1 and TRBC2, and exon 3 of TIGIT. White boxes represent untranslated regions, yellow boxes represent coding regions, and black boxes represent stop codons. [Figure 36] Evaluation of editing efficiency induced by sgRNAs used with CRISPR / Cas9 or a base editing system. Indel frequencies (A, left) and cytosine base conversions (A, right) produced by sgRNAs targeting TRAC, TRBC1 / 2, and TIGIT. Frequency of bystander cytokine modifications at target loci (B-C). CD3 and TIGIT protein expression after simultaneous disruption of TCR and TIGIT by sgRNA coupled with BE4max (D). Data are expressed as mean ± SEM. n=3. [Figure 37-1] Evaluation of off-target events at the genome level. (A) A scheme for two possible off-target events and strategies for investigating them. [Figure 37-2] Evaluation of off-target events at the genome level. (B) Experimental workflow. [Figure 38] Evaluation of sgRNA-independent off-target editing. (A) Percentage of nucleotide conversions in the predicted target gene and off-target sites. Data are expressed as mean ± SEM. n=3. (BC) (B) Variant type frequency and (C) Specific nucleotide conversion, measured by ultra-deep whole-exome sequencing. n=4. [Figure 39]Efficiency of ENTPD1 disruption. (A) Scheme of guide RNA designed to target ENTPD1. White boxes represent untranslated regions, yellow boxes represent coding regions, and black boxes represent stop codons. (B) Target cytosine base conversions resulting from C1, C2, and C3 coupled with BE4max. n=3. [Figure 40] Elimination of tumor cell lines. Elimination of tumor cell lines by TCR-edited T cells redirected to a specific tumor antigen (Ag). Data are expressed as mean ± SEM. n=3. [Figure 41-1] Elimination of primary CRC patient-derived organoids (PDOs). Elimination of primary CRC patient-derived organoids (PDOs) by TCR-edited T cells redirected to specific tumor antigens. Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 41-2] Elimination of primary CRC patient-derived organoids (PDOs). Elimination of primary CRC patient-derived organoids (PDOs) by TCR-edited T cells redirected to specific tumor antigens. Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 42-1] Elimination of organoids (PDOs) derived from CRC liver metastases by TCR-edited T cells redirected to specific tumor antigens. Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 42-2] Elimination of organoids (PDOs) derived from CRC liver metastases by TCR-edited T cells redirected to specific tumor antigens. Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 43]Elimination of primary PDAC patient-derived organoids (PDOs). Elimination of primary PDAC patient-derived organoids (PDOs) by TCR-edited T cells redirected to specific tumor antigens. Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 44] The effect of TIGIT disruption on the ability of MET-specific T cells to eliminate primary CRC patient-derived organoids (PDOs). TIGIT disruption improves the ability of MET-specific T cells to eliminate primary CRC patient-derived organoids (PDOs). Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 45] The effect of TIGIT disruption on the ability of DKK1-specific T cells to eliminate patient-derived organoids (PDOs) from clinical recombination cancer (CRC). TIGIT disruption improves the ability of DKK1-specific T cells to eliminate (A) primary and (B) metastatic CRC patient-derived organoids (PDOs). Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 46] The effect of TIGIT disruption on the ability of DKK1-specific T cells to eliminate (A) CRC liver metastases and (B) patient-derived organoids (PDOs) from primary PDAC. TIGIT disruption improves the ability of DKK1-specific T cells to eliminate (A) CRC liver metastases and (B) patient-derived organoids (PDOs) from primary PDAC. Significance is calculated by a linear regression curve fitted to the observed results. Data are expressed as mean ± SEM. n=3. [Figure 47-1] TIGIT disruption enabling tumor growth control by MSLN-specific T cells in vivo. (AC)(A) Experimental workflow for CRC and (C) PDAC orthotopic mouse models. (BD)(B) Tumor growth dynamics measured by in vivo bioluminescence in the CRC model and (D) ultrasound imaging in the PDAC model. The PDO increase factor is calculated as follows: BLI on day X / BLI on day 0. Significance is calculated by a linear regression curve fitted to the observed results. [Figure 47-2]TIGIT disruption enabling tumor growth control by MSLN-specific T cells in vivo. (A-C) Experimental workflows for (A) CRC and (C) PDAC syngeneic mouse models. (B-D) Tumor growth kinetics measured by (B) in vivo bioluminescence in the CRC model and (D) ultrasound imaging in the PDAC model. The doubling rate of PDOs was calculated as follows: BLI on day X / BLI on day 0. Significance was calculated by a linear regression curve fitting the observed results. [Figure 47-3] TIGIT disruption enabling tumor growth control by MSLN-specific T cells in vivo. (A-C) Experimental workflows for (A) CRC and (C) PDAC syngeneic mouse models. (B-D) Tumor growth kinetics measured by (B) in vivo bioluminescence in the CRC model and (D) ultrasound imaging in the PDAC model. The doubling rate of PDOs was calculated as follows: BLI on day X / BLI on day 0. Significance was calculated by a linear regression curve fitting the observed results. [Figure 48] Non-crossreactivity of CTSG TCR against other peptides from the human proteome. (A and C) Lists of the sequences of wild-type (wt) and synthetic peptides used in the alanine scanning assay. (B and D) CTSG TCR-transduced T cells were co-cultured with (B) HLA-A*24:02+ or (D) HLA-C*07:02+ EBV-BLCLs pulsed with the cognate wild-type epitope (P7 or P56) or modified ligand peptides showing alanine substitution of each residue (at a concentration of 100 μM, left graph, or 1 μM, right graph). Degranulation activity in CD8+ T cells was used as the readout. The co-culture assay was set up at an effector-to-target ratio of 1:1. The data represent experiments performed using engineered T cells from two different healthy donors. CTSG: cathepsin G, EBV-BLCLs: Epstein-Barr virus-transformed B lymphoblastoid cell lines.

Modes for Carrying Out the Invention

[0077] Various preferred features and embodiments of the present invention are described herein as non-limiting examples. This disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar to or equivalent to those described herein may be used in the implementation or testing of embodiments of this disclosure. Those skilled in the art will understand that all features of the present invention disclosed herein can be combined without departing from the scope of the disclosed invention.

[0078] As used herein and in the appended claims, the singular forms "a," "an," and "the" refer to multiple subjects unless the context clearly indicates otherwise.

[0079] As used herein, the terms “comprising,” “comprises,” and “comprised of” are synonymous with “including” or “includes,” or “containing” or “contains,” and are comprehensive or non-exclusive, not excluding additional, unlisted members, elements, or processes. The terms “comprising,” “comprises,” and “comprised of” also include the term “consisting of.”

[0080] Numerical ranges include the number defining the range. As used herein, the term “about” means approximately, within that range, roughly, or around that range. Unless otherwise specified, any nucleic acid sequence is written from left to right in the 5' to 3' direction; amino acid sequences are written from left to right in the amino to carboxyl direction.

[0081] The publications discussed herein are provided solely for their disclosure prior to the filing date of this application. Nothing herein should be construed as acknowledging that such publications constitute prior art to the claims attached herein. All publications referenced herein are incorporated herein by reference.

[0082] T cell receptor (TCR) In one embodiment, the present invention provides a T cell receptor (TCR) that binds to an immunogenic peptide when presented by a major histocompatibility complex (MHC). The present invention also provides an α-chain or β-chain of such a T cell receptor.

[0083] As used herein, the term “T-cell receptor” (TCR) may refer to a molecule capable of recognizing peptides when presented by MHC molecules. TCRs may be found on the surface of T cells and are responsible for recognizing antigen peptides bound to MHC molecules. During antigen processing, the antigen is degraded within the cell, and the antigen peptide is then transported to the cell surface by MHC molecules. The binding of the TCR to the peptide:MHC complex results in the activation of T lymphocytes on which the TCR is expressed, through a series of biochemical events mediated by associated enzymes, co-receptors, and specialized accessory molecules.

[0084] TCR affinity for peptides is determined by the association rate (k on ) and dissociation rate (k offThe TCR affinity can be determined by ( ) and expressed as the equilibrium dissociation constant (KD). The TCR affinity can be determined by any preferred method, e.g., titration calorimetry, surface plasmon resonance (SPR), or fluorescence microscopy (see, e.g., Piepenbrink, KH et al., 2009. Methods Enzymol., 466, pp:359-381). When presented by MHC, the TCR can bind to the peptide with affinities of about ≤100 μM, ≤50 μM, ≤40 μM, ≤30 μM, ≤20 μM, or ≤10 μM. When presented by MHC, the TCR can bind to the peptide with affinities of about 0.1 μM to ≤100 μM, ≤1 μM to ≤100 μM, or ≤5 μM to ≤100 μM. When presented by MHC, TCRs can bind to peptides with affinities of approximately 0.1 μM to 10 μM, 1 μM to 10 μM, or 5 μM to 10 μM.

[0085] Naturally occurring TCR heterodimers consist of alpha (α) and beta (β) chains in approximately 95% of T cells, while about 5% of T cells have TCRs consisting of gamma (γ) and delta (δ) chains. Each chain of the natural TCR is a member of the immunoglobulin superfamily and has an N-terminal immunoglobulin (Ig) variable (V) region, an Ig constant (C) region, a transmembrane / transmembrane region, and a short cytoplasmic tail at the C-terminus.

[0086] The TCR of the present invention may be a heterodimer of two chains, an α chain and a β chain (or optionally a γ chain and a δ chain), or it may be a single-chain TCR construct. Preferably, when the TCR of the present invention is an α chain and a β chain heterodimer, the α chain and the β chain bind to the same immunogenic peptide. The α chain and β chain (or their variable domain or CDR) provided herein may be combined in any preferred combination. For example, if three α chains and three β chains (or their variable domain or CDR) that bind to the same immunogenic peptide are provided, the TCR of the present invention may include combinations of α1 / β1, α1 / β2, α1 / β3, α2 / β1, α2 / β2, α2 / β3, α3 / β1, α3 / β2, or α3 / β3. In some embodiments, the TCR of the present invention includes a combination of an α1 chain (or its variable domain or CDR) and a β1 chain (or its variable domain or CDR). In some embodiments, the TCR of the present invention includes a combination of an α1 chain (or its variable domain or CDR) and a β2 chain (or its variable domain or CDR). In some embodiments, the TCR of the present invention includes a combination of an α1 chain (or its variable domain or CDR) and a β3 chain (or its variable domain or CDR). In some embodiments, the TCR of the present invention includes a combination of an α2 chain (or its variable domain or CDR) and a β1 chain (or its variable domain or CDR). In some embodiments, the TCR of the present invention includes a combination of an α2 chain (or its variable domain or CDR) and a β2 chain (or its variable domain or CDR). In some embodiments, the TCR of the present invention includes a combination of an α2 chain (or its variable domain or CDR) and a β3 chain (or its variable domain or CDR). In some embodiments, the TCR of the present invention includes a combination of an α3 chain (or its variable domain or CDR) and a β1 chain (or its variable domain or CDR). In some embodiments, the TCR of the present invention includes a combination of an α3 chain (or its variable domain or CDR) and a β2 chain (or its variable domain or CDR).In some embodiments, the TCR of the present invention includes a combination of an α3 chain (or its variable domain or CDR) and a β3 chain (or its variable domain or CDR).

[0087] Variable domains can determine the specificity of the TCR. Both the α and β chains of the TCR have three hypervariable regions or complementarity-determining regions (CDRs). The α or β chain, for example, contains CDR1, CDR2, and CDR3 in the order from the amino terminus to the carboxyl terminus. Generally, CDR3 is the primary CDR responsible for the recognition of the processed antigen, while CDR1 of the α chain has also been shown to interact with the N-terminal portion of the antigenic peptide, while CDR1 of the β chain interacts with the C-terminal portion of the peptide. CDR2 is thought to recognize MHC molecules. Preferred variable domains are described herein.

[0088] The TCR of the present invention may include a constant domain. The constant domain may allow the TCR to associate with other molecules such as CD3 and ζ chains, which have three distinct chains (γ, δ, and ε) in mammals. These accessory molecules have negatively charged transmembrane regions and are essential for signal propagation from the TCR to the cell. The CD3 and ζ chains, together with the TCR, form what is known as the T cell receptor complex.

[0089] The constant domain may consist of short linking sequences in which cysteine ​​residues form disulfide bonds, creating a link between the two chains. The constant domain may include an Ig constant (C) region, a transmembrane / transcellular domain, and a cytoplasmic tail.

[0090] The α-chain of the TCR of the present invention may include a constant domain encoded by the TRAC gene. The α-chain constant domain may have the amino acid sequence described in UniProt entry P01848. An example of an α-chain constant domain has the amino acid sequence shown in Sequence ID No. 1 below.

[0091] The TCR of the present invention may include or consist of an α-chain constant domain comprising the amino acid sequence of SEQ ID NO: 1 or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0092]

Table 1

[0093] The β-chain of the TCR of the present invention may include a constant domain encoded by the TRBC1 or TRBC2 gene. The β-chain constant domain may have the amino acid sequence described in UniProt entry P01850 or A0A5B9. An exemplary β-chain constant domain encoded by the TRBC1 gene has the amino acid sequence shown in SEQ ID NO: 2 below. An exemplary β-chain constant domain encoded by the TRBC2 gene has the amino acid sequence shown in SEQ ID NO: 3 below.

[0094] The TCR of the present invention may include a β-chain constant domain comprising the amino acid sequence of SEQ ID NO: 2 or 3 or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0095]

Table 2

[0096] The TCR of the present invention may have one or more additional cysteine residues in each of the α-chain and β-chain such that the TCR may include two or more disulfide bonds in the constant domain.

[0097] Signaling from T cell complexes can be enhanced by the co-binding of MHC molecules by specific co-receptors. For helper T cells, this co-receptor may be CD4 (specific to class II MHC), while for cytotoxic T cells, it may be CD8 (specific to class I MHC). The co-receptor enables long-term binding between antigen-presenting cells and T cells and can recruit essential intracellular molecules (e.g., LCK) involved in signal transduction by activated T lymphocytes.

[0098] The TCR of the present invention may be a hybrid TCR containing sequences derived from more than one species. For example, mouse TCRs have been found to be expressed more efficiently in human T cells than human TCRs. Therefore, the TCR may include a human variable domain and a mouse sequence within a constant domain. A drawback of this approach is that the mouse constant sequence may induce an immune response, leading to rejection of the transplanted T cells. However, the pre-treatment regimens used to prepare patients for adoptive T cell therapy may provide sufficient immunosuppression to allow engraftment of T cells expressing the mouse sequence.

[0099] The TCR of the present invention may be, for example, a soluble TCR in which one or more constant domains are omitted or modified. Other suitable methods for manipulating soluble TCRs are known in the art (see, for example, Robinson, RA et al., 2021. The FEBS Journal, 288(21), pp. 6159-6173).

[0100] The term "soluble TCR" refers to TCRs that are not fixed to the cell surface. Soluble TCRs typically contain at least a portion of the variable domain and constant domain, but lack the transmembrane domain and intracellular cytoplasmic domain. Soluble TCRs may contain stabilizing mutations and / or modifications (e.g., point mutations, interchain disulfide bonds and / or peptide linkers). Such modifications are known in the art (e.g., Robinson, RA et al., 2021. The FEBS Journal, 288(21), pp. 6159-6173).

[0101] In some embodiments, the TCR comprises one or more mutations for the removal of one or more N-glycosylation sites. Preferably, the N-glycosylation sites are located in the TCR constant domain. Deletion of an N-glycosylation site in the TCR constant domain is described in Kuball, J et al., (2009) J Exp Med 206:463-75. In some embodiments, one or more mutations are substitutions of amino acid N to amino acid Q in the NXS / T motif. For example, the substitution may be located at one or more of the following positions in the TCRα constant gene: position 36, 90, or 109, and / or position 85 or 6 in the TCRβ constant gene. Preferably, the substitution is located at position 36 in the TCRα constant gene. Mutations of the TCR constant domain disclosed herein may be described based on the numbering convention in which the first amino acid of each of SEQ ID NOs. 1-3 is assigned to position 2.

[0102] Complementarity Determination Area (CDR) The diversity of T cell receptors is concentrated in the CDR3 region, which is primarily involved in antigen recognition. The sequence of the CDR3 region of the TCR of the present invention may be selected from those described herein. The TCR of the present invention may include a CDR comprising the CDR3α and CDR3β pairs described herein or a CDR comprising them.

[0103] The portion of the TCR that establishes the majority of contact with antigen peptides bound to the major histocompatibility complex (MHC) is the complementarity-determining region 3 (CDR3), which is unique to each T cell clone. The CDR3 region is generated during somatic cell rearrangement events that occur in the thymus and contains discontinuous genes belonging to variable (V), diverse (for D, β, and δ chains) and linked (J) genes. Furthermore, random nucleotides inserted / deleted at the rearrangement locus of each TCR chain gene greatly increase the diversity of the highly variable CDR3 sequence. Therefore, the frequency of a particular CDR3 sequence in a biological sample indicates the abundance of a particular T cell population. The large diversity of the TCR repertoire in healthy humans provides broad protection against various foreign antigens presented by MHC molecules on the surface of antigen-presenting cells. In this regard, theoretically, 10 15 It should be noted that different TCRs can be generated in the thymus.

[0104] The sequences of the CDR1 and CDR2 regions of the TCR of the present invention may also be selected from those described herein. The TCR of the present invention may include or comprise a CDR comprising the CDR1α, CDR2α, CDR3α, CDR1β, CDR2β, and CDR3β sets described herein.

[0105] A CDR may include one, two, or three substitutions, additions, or deletions from a given sequence, provided that, for example, the TCR retains its ability to bind to the corresponding immunogenic peptide when presented by an MHC molecule. In some embodiments, each CDR3 includes up to three substitutions, additions, or deletions, up to two substitutions, additions, or deletions, up to one substitution, addition, or deletion, or any combination thereof. In some embodiments, each CDR3 includes up to three substitutions, up to two substitutions, up to one substitution, or any combination thereof. In some embodiments, each CDR includes up to three substitutions, additions, or deletions, up to two substitutions, additions, or deletions, up to one substitution, addition, or deletion, or any combination thereof. In some embodiments, each CDR includes up to three substitutions, up to two substitutions, up to one substitution, or any combination thereof.

[0106] major histocompatibility complex (MHC) molecules Typically, TCRs bind to peptides as part of a peptide:MHC complex. The TCRs of the present invention can bind to MHC I and / or MHC II peptide complexes. The MHC peptide complexes may be present on the surface of antigen-presenting cells such as dendritic cells or B cells, or any other cells including cancer cells, or they may be immobilized, for example, by coating them onto beads or plates.

[0107] The human leukocyte antigen system (HLA) is the name given to the gene complex that codes for MHC in humans, and includes HLA class I antigens (A, B, and C) and HLA class II antigens (DP, DQ, and DR). HLA alleles A, B, and C mainly present peptides derived from intracellular proteins, such as proteins expressed within cells. The TCR is limited to human leukocyte antigen (HLA) alleles.

[0108] During T cell development in vivo, T cells undergo a positive selection process to ensure recognition of self-MHC, followed by a negative process to eliminate T cells that bind too strongly to MHC presenting self-antigens. As a result, certain T cells and the TCRs they express recognize only peptides presented by certain MHC molecules, i.e., those encoded by specific HLA alleles. This is known as HLA constraint. Preferred MHC binding motifs for different HLA alleles are listed in The Immune Epitope Database (IEDB) (Vita, R. et al., 2019. Nucleic acids research, 47(D1), pp. D339-D343).

[0109] Immunogenic peptides The TCR of the present invention binds to immunogenic peptides when presented by MHC.

[0110] As used herein, the term "protein" may include single-chain polypeptide molecules, as well as multiple polypeptide complexes in which individual constituent polypeptides are linked by covalent or non-covalent means.

[0111] As used herein, the term "polypeptide" may refer to a polymer in which monomers are amino acids and are linked to one another via peptide bonds or disulfide bonds.

[0112] As used herein, the term "peptide" may refer to a group of amino acid residues linked by peptide bonds. A peptide may consist of fewer than 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 amino acid residues. A peptide may consist of 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more amino acid residues. Preferably, a peptide is 5-20 amino acid long, 8-15 amino acid residue long, 9-14 amino acid residue long, 9-13 amino acid residue long, 9-12 amino acid residue long, 9-11 amino acid residue long, or 9-10 amino acid residue long. In some embodiments, the peptide of the present invention is 9 amino acid residue long.

[0113] As used herein, the terms “immunogenic peptide” or “immunogenic fragment” may refer to a peptide that can be presented by an MHC molecule and subsequently recognized by a TCR. Immunogenic peptides or fragments may be derived, for example, from AML-related antigens, CRC-related antigens, or PDAC-related antigens. Immunogenic peptides or fragments may consist of fewer than 30, fewer than 25, fewer than 20, fewer than 19, fewer than 18, fewer than 17, fewer than 16, fewer than 15, fewer than 14, fewer than 13, fewer than 12, fewer than 11, or fewer than 10 amino acid residues. Immunogenic peptides or fragments may consist of 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more amino acid residues. Preferably, the immunogenic peptide or fragment has a length of 5-20 amino acids, 8-15 amino acid residues, 9-14 amino acid residues, 9-13 amino acid residues, 9-12 amino acid residues, 9-11 amino acid residues, or 9-10 amino acid residues. In some embodiments, the immunogenic peptide or immunogenic fragment of the present invention has a length of 9 amino acid residues.

[0114] The immunogenic peptides of the present invention may be derived from any suitable human protein, for example, a tumor-associated antigen. As used herein, “tumor-associated antigen” may refer to an antigen protein produced in tumor cells. In some embodiments, the immunogenic peptides of the present invention are derived from AML-associated antigens, CRC-associated antigens, and / or PDAC-associated antigens.

[0115] In some embodiments, the immunogenic peptides of the present invention are derived from AML-related antigens. As used herein, “AML-related antigen” may refer to antigenic proteins produced in acute myeloid leukemia cells. AML-related antigens may act as useful tumor markers and as potential candidates for use in immunotherapy. AML-related antigens may include PR3 / NE, EZH2, PRAME, CTSG, and CCNA1.

[0116] In some embodiments, the immunogenic peptides of the present invention are derived from CRC-related antigens and / or PDAC-related antigens. As used herein, “CRC-related antigen” may refer to an antigen protein produced in colorectal cancer cells. CRC-related antigens may act as useful tumor markers and as potential candidates for use in immunotherapy. As used herein, “PDAC-related antigen” may refer to an antigen protein produced in pancreatic ductal adenocarcinoma cells. PDAC-related antigens may act as useful tumor markers and as potential candidates for use in immunotherapy. CRC-related antigens and / or PDAC-related antigens may include HER2, DKK1, MUC5AC, AGR2, c-MET, MSLN, CTDP1, TACC2, or PTGFRN.

[0117] Various uses of the peptides described herein are envisioned by the present invention. For example, the peptides described herein may be administered to a subject, e.g., a human subject. Administration of the peptides of the present invention may induce an immune response against cells expressing or overexpressing the corresponding protein; that is, the peptides may be immunogenic peptides. The peptides described herein may be used to screen and / or identify novel TCR sequences. For example, T2 cells may be pulsed with the peptides described herein and incubated with a population of T cells isolated from a donor. In this approach, the expression of cytokines, e.g., CD107a and IFNγ, may serve as indicators of T cells that recognize the peptides.

[0118] HER2-specific TCR In one embodiment, the present invention provides a TCR that binds to the HER2 peptide when presented by MHC.

[0119] HER2 HER2 is a protein encoded by the ERBB2 gene and is also known as human epidermal growth factor receptor 2, receptor tyrosine protein kinase erbB-2, or CD340.

[0120] HER2 is a member of the epidermal growth factor receptor family with tyrosine kinase activity. Receptor dimerization leads to autophosphorylation of tyrosine residues within the receptor's cytoplasmic domain, initiating various signaling pathways that result in cell proliferation and tumorigenesis. Amplification or overexpression of HER2 occurs in approximately 15–30% of breast cancers and 10–30% of gastric / esophageal cancers. HER2 overexpression has also been observed in other cancers such as ovarian, endometrial, bladder, lung, colon, and head and neck cancers (see, e.g., Iqbal, N. and Iqbal, N., 2014. Molecular Biology International, p. 852748).

[0121] The HER2 protein may have the amino acid sequence described in UniProt entry P04626. An exemplary HER2 protein has the amino acid sequence shown in Sequence ID No. 4 below.

[0122] [Table 3]

[0123] HER2 peptide As used herein, the term "HER2 peptide" may refer to a peptide comprising an amino acid sequence derived from the HER2 protein. Preferably, the HER2 peptide may comprise at least five, at least six, at least seven, at least eight, or at least nine consecutive amino acid residues of the HER2 protein amino acid sequence (or a variant thereof). In some embodiments, the HER2 peptide comprises nine consecutive amino acid residues of the HER2 protein amino acid sequence (or a variant thereof).

[0124] The HER2 peptide may be an immunogenic peptide. The HER2 peptide may bind to any suitable MHC. Preferably, the HER2 peptide binds to an MHC encoded by an HLA class I allele or an HLA class II allele. Preferably, the HER2 peptide binds to an MHC encoded by an HLA-A allele. Preferably, the HER2 peptide binds to an MHC encoded by an HLA-A allele. * It binds to the MHC encoded by the 02 allele. Preferably, the HER2 peptide is HLA-A * It binds to the MHC encoded by 0201.

[0125] An exemplary HER2 peptide has the amino acid sequence shown in Sequence ID No. 5 below. KIFGSLAFL Exemplary HER2 peptide (SEQ ID NO: 5)

[0126] The HER2 peptide may include, or consist of, the amino acid sequence of SEQ ID NO: 5, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the HER2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 5, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the HER2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 5, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the HER2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 5, or a variant thereof having up to three amino acid substitutions. In some embodiments, the HER2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 5, or a variant thereof having up to two amino acid substitutions. In some embodiments, the HER2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 5, or a variant thereof having up to one amino acid substitution. In some embodiments, the HER2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 5.

[0127] HER2-specific TCR sequence The inventors determined the amino acid sequence of the HER2-specific TCR, which includes the CDR region involved in binding specificity to the HER2 peptide.

[0128] A HER2-specific TCR may be constrained to any suitable MHC (e.g., one or more HLA alleles). Preferably, the HER2-specific TCR is constrained to an HLA class I or HLA class II allele. Preferably, the HER2-specific TCR is constrained to an HLA-A allele. Preferably, the HER2-specific TCR is constrained to an HLA-A allele. * It is bound to the O2 allele. Preferably, the HER2-specific TCR is HLA-A * He was detained at 0201.

[0129] Examples of HER2-specific TCR amino acid sequences are provided in the table below.

[0130] [Table 4]

[0131] In one embodiment, the present invention provides a TCR comprising CDR3α, which comprises or consists of the amino acid sequence of SEQ ID NO: 8, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or CDR3β, which comprises the amino acid sequence of SEQ ID NO: 13, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0132] In some embodiments, the CDR3 variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR3 variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR3 variant has up to three amino acid substitutions. In some embodiments, the CDR3 variant has up to two amino acid substitutions. In some embodiments, the CDR3 variant has up to one amino acid substitution.

[0133] In some embodiments, the TCR comprises CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 8, and / or CDR3β containing or consisting of the amino acid sequence of SEQ ID NO: 13.

[0134] In some embodiments, the TCR further comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 6, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 7, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 11, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 12, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0135] In some embodiments, the TCR includes an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 9, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 14, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0136] In some embodiments, the TCR includes an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 10, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 15 or 16, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0137] In one embodiment, the present invention provides a TCR comprising (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 6, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 7, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 8, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 11, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 12, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 13, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0138] In some embodiments, the CDR variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR variant has up to three amino acid substitutions. In some embodiments, the CDR variant has up to two amino acid substitutions. In some embodiments, the CDR variant has up to one amino acid substitution.

[0139] In some embodiments, the TCR comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 6, (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 7, (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 8, and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 11, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 12, and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 13.

[0140] In one embodiment, the present invention provides a TCR comprising an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 9, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 14, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0141] In one embodiment, the present invention relates to an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 9, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises an α-chain variable domain including CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 8, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or The present invention provides a TCR comprising a β-chain variable domain comprising or having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises a β-chain variable domain containing CDR3β comprising or having at least 3 amino acid substitutions, additions, or deletions thereto, the amino acid sequence of SEQ ID NO: 13, or a variant thereof.

[0142] In one embodiment, the present invention relates to an α-chain variable domain comprising or comprising the amino acid sequence of SEQ ID NO: 9, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises (a) CDR1α comprising or comprising the amino acid sequence of SEQ ID NO: 6, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising the amino acid sequence of SEQ ID NO: 7, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising the amino acid sequence of SEQ ID NO: 8, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or The present invention provides a β-chain variable domain comprising, or comprising, the amino acid sequence of SEQ ID NO: 14, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises: (a) CDR1β comprising or comprising the amino acid sequence of SEQ ID NO: 11, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or comprising the amino acid sequence of SEQ ID NO: 12, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising the amino acid sequence of SEQ ID NO: 13, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0143] In one embodiment, the present invention provides a TCR comprising an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 10, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain comprising the amino acid sequence of SEQ ID NO: 15 or 16, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0144] In one embodiment, the present invention relates to an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 10, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 8, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or The present invention provides a TCR comprising a β-chain containing or comprising the amino acid sequence of sequence number 15 or 16, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain contains CDR3β, which contains or comprises the amino acid sequence of sequence number 13, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0145] In one embodiment, the present invention relates to an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 10, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 6, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 7, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3α comprising the amino acid sequence of SEQ ID NO: 8, or a variant thereof having up to three amino acid substitutions, additions, or deletions. The present invention provides a TCR comprising a β-chain containing or comprising the amino acid sequence of sequence 15 or 16, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises (a) CDR1β containing or comprising the amino acid sequence of sequence number 11, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β containing or comprising the amino acid sequence of sequence number 12, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β containing or comprising the amino acid sequence of sequence number 13, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0146] PR3 / NE-specific TCR In one embodiment, the present invention provides a TCR that binds to a PR3 / NE peptide when presented by an MHC.

[0147] PR3 and NE PR3 is a protein encoded by the PRTN3 gene and is also known as proteinase 3 or myeloblast. NE is a protein encoded by the ELANE gene and is also known as neutrophil elastase or leukocyte elastase.

[0148] PR3 and NE are both serine proteases localized in the primary azurophilic granules of neutrophils, and their primary function is to combat bacterial infection by processing pathogenic proteins into smaller peptides both intracellularly and extracellularly. Their overexpression has been observed in the cytoplasm of leukemic blasts and leukemic stem cells in primary specimens, making them attractive antigens to be selected for adoptive cell therapy (ACT) for acute myeloid leukemia (AML) (Greiner, J. et al., 2008. Clinical Cancer Research, 14(22), pp.7161-7166). Furthermore, tumor cell lines negative for PR3 and NE expression have been shown to internalize these antigens from exogenous sources and cross-present them (Alatrash, G., et al., 2018. Oncotarget, 9(4), p.4280). This internalization mechanism is interesting in ACT because it expands the range of cancer types that are sensitive to immunotherapy against PR3 and NE.

[0149] The PR3 protein may have the amino acid sequence described in UniProt entry P24158. An exemplary PR3 protein has the amino acid sequence shown in SEQ ID NO: 17 below.

[0150] [Table 5]

[0151] NE proteins may have the amino acid sequence described in UniProt entry P08246. An exemplary NE protein has the amino acid sequence shown in SEQ ID NO: 18 below.

[0152] [Table 6]

[0153] PR3 / NE peptide As used herein, the term "PR3 / NE peptide" can refer to a peptide comprising an amino acid sequence derived from the PR3 protein and / or the NE protein. Preferably, the PR3 / NE peptide can comprise at least 5, at least 6, at least 7, at least 8, or at least 9 consecutive amino acid residues of the PR3 protein amino acid sequence (or a variant thereof) and / or the NE protein amino acid sequence (or a variant thereof). In some embodiments, the PR3 / NE peptide comprises 9 consecutive amino acid residues of the PR3 protein amino acid sequence (or a variant thereof) and / or the NE protein amino acid sequence (or a variant thereof).

[0154] In some embodiments, the PR3 / NE peptide comprises at least 5, at least 6, at least 7, at least 8, or at least 9 consecutive amino acid residues of the PR3 protein amino acid sequence (or a variant thereof) and the NE protein amino acid sequence (or a variant thereof). In some embodiments, the PR3 / NE peptide comprises at least 9 consecutive amino acid residues of the PR3 protein amino acid sequence (or a variant thereof) and the NE protein amino acid sequence (or a variant thereof).

[0155] The PR3 / NE peptide can be an immunogenic peptide. The PR3 / NE peptide can bind to any suitable MHC. Preferably, the PR3 / NE peptide binds to an MHC encoded by an HLA class I allele or an HLA class II allele. Preferably, the PR3 / NE peptide binds to an MHC encoded by an HLA-A allele. Preferably, the PR3 / NE peptide binds to an MHC encoded by HLA-A * 02 allele. Preferably, the PR3 / NE peptide binds to an MHC encoded by HLA-A * 0201.

[0156] An exemplary PR3 / NE peptide has the amino acid sequence shown in SEQ ID NO: 19 below. VLQELNVTV Example PR3 / NE peptide (SEQ ID NO: 19)

[0157] The PR3 / NE peptide may include, or consist of, the amino acid sequence of SEQ ID NO: 19, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the PR3 / NE peptide includes or consists of the amino acid sequence of SEQ ID NO: 19, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the PR3 / NE peptide includes or consists of the amino acid sequence of SEQ ID NO: 19, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the PR3 / NE peptide includes or consists of the amino acid sequence of SEQ ID NO: 19, or a variant thereof having up to three amino acid substitutions. In some embodiments, the PR3 / NE peptide includes or consists of the amino acid sequence of SEQ ID NO: 19, or a variant thereof having up to two amino acid substitutions. In some embodiments, the PR3 / NE peptide includes or consists of the amino acid sequence of SEQ ID NO: 19, or a variant thereof having up to one amino acid substitution. In some embodiments, the PR3 / NE peptide includes or consists of the amino acid sequence of SEQ ID NO: 19.

[0158] PR3 / NE-specific TCR sequence The inventors have determined the amino acid sequence of the PR3 / NE-specific TCR, which includes the CDR region and is involved in binding specificity to the PR3 / NE peptide.

[0159] The PR3 / NE-specific TCR can be constrained to any suitable MHC (e.g., one or more HLA alleles). Preferably, the PR3 / NE-specific TCR is constrained to an HLA class I or HLA class II allele. Preferably, the PR3 / NE-specific TCR is constrained to an HLA-A allele. Preferably, the PR3 / NE-specific TCR is constrained to an HLA-A allele. * It is constrained to the 02 allele. Preferably, the PR3 / NE specific TCR is HLA-A * He was detained at 0201.

[0160] Examples of PR3 / NE-specific TCR amino acid sequences are provided in the table below.

[0161] [Table 7]

[0162] In one embodiment, the present invention provides a TCR comprising CDR3α, which comprises or consists of the amino acid sequence of SEQ ID NO: 22, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or CDR3β, which comprises or consists of the amino acid sequence of SEQ ID NO: 27, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0163] In some embodiments, the CDR3 variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR3 variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR3 variant has up to three amino acid substitutions. In some embodiments, the CDR3 variant has up to two amino acid substitutions. In some embodiments, the CDR3 variant has up to one amino acid substitution.

[0164] In some embodiments, the TCR comprises CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 22, and / or CDR3β containing or consisting of the amino acid sequence of SEQ ID NO: 27.

[0165] In some embodiments, the TCR further comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 20, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 21, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 25, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 26, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0166] In some embodiments, the TCR includes an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 23, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 28, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0167] In some embodiments, the TCR includes an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 24, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 29 or 30, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0168] In one embodiment, the present invention provides a TCR comprising (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 20, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 21, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 22, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 25, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 26, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 27, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0169] In some embodiments, the CDR variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR variant has up to three amino acid substitutions. In some embodiments, the CDR variant has up to two amino acid substitutions. In some embodiments, the CDR variant has up to one amino acid substitution.

[0170] In some embodiments, the TCR comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 20, (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 21, (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 22, and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 25, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 26, and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 27.

[0171] In one embodiment, the present invention provides a TCR comprising an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 23, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 28, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0172] In one embodiment, the present invention relates to an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 23, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises an α-chain variable domain including CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 22, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or The present invention provides a TCR comprising a β-chain variable domain that includes or comprises the amino acid sequence of SEQ ID NO: 28, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises CDR3β, which includes the amino acid sequence of SEQ ID NO: 27, or a variant thereof having up to three amino acid substitutions, additions, or deletions thereto.

[0173] In one embodiment, the present invention relates to an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 23, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 20, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising the amino acid sequence of SEQ ID NO: 21, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising the amino acid sequence of SEQ ID NO: 22, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or The present invention provides a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 28, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises: (a) CDR1β comprising the amino acid sequence of SEQ ID NO: 25, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising the amino acid sequence of SEQ ID NO: 26, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising the amino acid sequence of SEQ ID NO: 27, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0174] In one embodiment, the present invention provides a TCR comprising an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 24, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain comprising the amino acid sequence of SEQ ID NO: 29 or 30, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0175] In one embodiment, the present invention relates to an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 24, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 22, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or The present invention provides a TCR comprising a β-chain comprising the amino acid sequence of sequence number 29 or 30, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises a CDR3β comprising the amino acid sequence of sequence number 27, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0176] In one embodiment, the present invention relates to an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 24, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 20, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 21, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 22, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or The present invention provides a TCR comprising a β-chain containing or comprising the amino acid sequence of sequence number 29 or 30, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises (a) CDR1β containing or comprising the amino acid sequence of SEQ ID NO: 25, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β containing or comprising the amino acid sequence of SEQ ID NO: 26, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β containing or comprising the amino acid sequence of SEQ ID NO: 27, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0177] EZH2 / PRAME-specific TCR In one embodiment, the present invention provides a TCR that, when presented by an MHC, binds to an EZH2 peptide or a PRAME peptide.

[0178] EZH2 EZH2 is a protein encoded by the EZH2 gene and is also known as the histone-lysine N-methyltransferase EZH2 or as an enhancer of zeste homolog 2.

[0179] EZH2 is a lysine methyltransferase of Polycomb repression complex 2 (PRC2) that catalyzes the addition of three methyl groups to lysine 27 (K27) of histone 3 (H3), and thus establishes and maintains the H3K27 trimethylation repression mark (Li, B. and Chng, WJ, 2019. Journal of Hematology & Oncology, 12, pp. 1-13). Under physiological conditions, it acts as a tumor suppressor gene, but upon oncogenic hits such as gain-of-function mutations in the EZH2 gene or its genomic regulators (mRNA or lncRNA), the EZH2 protein can be overexpressed, as shown in various hematological malignancies. However, loss-of-function mutations can also occur in the EZH2 gene, which result in loss of the trimethylation repression mark and expression of target genes, driving the development of acute myeloid leukemia (AML) and associated with a poor prognosis.

[0180] The EZH2 protein may have the amino acid sequence described in UniProt entry Q15910. An exemplary EZH2 protein has the amino acid sequence shown in SEQ ID NO: 31 below.

[0181] [Table 8]

[0182] EZH2 peptide As used herein, the term "EZH2 peptide" may refer to a peptide comprising an amino acid sequence derived from the EZH2 protein. Preferably, the EZH2 peptide may comprise at least five, at least six, at least seven, at least eight, or at least nine consecutive amino acid residues of the EZH2 protein amino acid sequence (or a variant thereof). In some embodiments, the EZH2 peptide comprises nine consecutive amino acid residues of the EZH2 protein amino acid sequence (or a variant thereof).

[0183] The EZH2 peptide may be an immunogenic peptide. The EZH2 peptide can bind to any suitable MHC. Preferably, the EZH2 peptide binds to an MHC encoded by an HLA class I allele or an HLA class II allele. Preferably, the EZH2 peptide binds to an MHC encoded by an HLA-A allele. Preferably, the EZH2 peptide binds to an MHC encoded by an HLA-A allele. * It binds to the MHC encoded by the O2 allele. Preferably, the EZH2 peptide binds to HLA-A * It binds to the MHC encoded by 0201.

[0184] An exemplary EZH2 peptide has the amino acid sequence shown in Sequence ID No. 32 below. KESRPPRKF Exemplary EZH2 peptide (SEQ ID NO: 32)

[0185] The EZH2 peptide may include, or consist of, the amino acid sequence of SEQ ID NO: 32, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the EZH2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 32, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the EZH2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 32, or a variant thereof having one amino acid substitution, addition, or deletion. In some embodiments, the EZH2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 32, or a variant thereof having up to three amino acid substitutions. In some embodiments, the EZH2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 32, or a variant thereof having up to two amino acid substitutions. In some embodiments, the EZH2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 32, or a variant thereof having one amino acid substitution. In some embodiments, the EZH2 peptide includes or consists of the amino acid sequence of SEQ ID NO: 32.

[0186] PRAME (Melanoma Priority Antigen) PRAME is a protein encoded by the PRAME gene and is known as the melanoma antigen, the antigen preferentially expressed in tumors, or the Opa-interacting protein 4.

[0187] PRAME belongs to the cancer-testis antigen (CTA) family of genes, which are highly expressed on testicular germ cells but absent in other healthy tissues. High levels of PRAME expression are detected on malignant cells, including most primary and metastatic melanomas. In its nuclear localization, PRAME interacts with EZH2 to suppress the retinoic acid receptor (RAR) signaling cascade, which is known to mediate tumor suppressor function. In fact, binding of retinoic acid (RA) to RAR in the nucleus induces RARβ2 gene expression and transcription of RA target genes that regulate cell cycle arrest and caspase-dependent apoptosis pathways in responsive cells. Therefore, inhibition of RAR signaling by PRAME may represent a significant contributing factor in the progression of acute myeloid leukemia (AML) disease (see, e.g., Wadelin, F. et al., 2010. Molecular Cancer, 9, pp. 1-10).

[0188] PRAME proteins may have the amino acid sequence shown in UniProt entry P78395. An exemplary PRAME protein has the amino acid sequence shown in SEQ ID NO: 33 below.

[0189] [Table 9]

[0190] PRAME peptide As used herein, the term "PRAME peptide" may refer to a peptide comprising an amino acid sequence derived from the PRAME protein. Preferably, the PRAME peptide may comprise at least five, at least six, at least seven, at least eight, or at least nine consecutive amino acid residues of the PRAME protein amino acid sequence (or a variant thereof). In some embodiments, the PRAME peptide comprises nine consecutive amino acid residues of the PRAME protein amino acid sequence (or a variant thereof).

[0191] PRAME peptides may be immunogenic peptides. PRAME peptides can bind to any suitable MHC. Preferably, PRAME peptides bind to MHC encoded by an HLA class I allele or an HLA class II allele. Preferably, PRAME peptides bind to MHC encoded by an HLA-A allele. Preferably, PRAME peptides bind to MHC encoded by an HLA-A allele. * It binds to the MHC encoded by the O2 allele. Preferably, the PRAME peptide is HLA-A * It binds to the MHC encoded by 0201.

[0192] An exemplary PRAME peptide has the amino acid sequence shown in SEQ ID NO: 34 below. ALYVDSLFFL Exemplary PRAME peptide (SEQ ID NO: 34)

[0193] The PRAME peptide may contain or consist of the amino acid sequence of SEQ ID NO: 34, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the PRAME peptide contains or consists of the amino acid sequence of SEQ ID NO: 34, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the PRAME peptide contains or consists of the amino acid sequence of SEQ ID NO: 34, or a variant thereof having one amino acid substitution, addition, or deletion. In some embodiments, the PRAME peptide contains or consists of the amino acid sequence of SEQ ID NO: 34, or a variant thereof having up to three amino acid substitutions. In some embodiments, the PRAME peptide contains or consists of the amino acid sequence of SEQ ID NO: 34, or a variant thereof having up to two amino acid substitutions. In some embodiments, the PRAME peptide contains or consists of the amino acid sequence of SEQ ID NO: 34, or a variant thereof having one amino acid substitution. In some embodiments, the PRAME peptide contains or consists of the amino acid sequence of SEQ ID NO: 34.

[0194] EZH2 / PRAME-specific TCR sequence The inventors have determined the amino acid sequence of an EZH2 / PRAME-specific TCR that includes a CDR region involved in binding specificity to EZH2 or PRAME peptides.

[0195] The EZH2 / PRAME-specific TCR can be constrained to any suitable MHC (e.g., one or more HLA alleles). Preferably, the EZH2 / PRAME-specific TCR is constrained to an HLA class I or HLA class II allele. Preferably, the EZH2 / PRAME-specific TCR is constrained to an HLA-A allele. Preferably, the EZH2 / PRAME-specific TCR is constrained to an HLA-A allele. * It is constrained to the O2 allele. Preferably, the EZH2 / PRAME specific TCR is HLA-A * He was detained at 0201.

[0196] Examples of EZH2 / PRAME-specific TCR amino acid sequences are shown in the table below.

[0197] [Table 10-1]

[0198] [Table 10-2]

[0199] In one embodiment, the present invention provides a TCR comprising (i) the amino acid sequence of SEQ ID NO: 37, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of SEQ ID NO: 42, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR3α comprising (i) the amino acid sequence of SEQ ID NO: 47, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of SEQ ID NO: 53, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR3β comprising

[0200] In some embodiments, the CDR3 variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR3 variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR3 variant has up to three amino acid substitutions. In some embodiments, the CDR3 variant has up to two amino acid substitutions. In some embodiments, the CDR3 variant has up to one amino acid substitution.

[0201] In some embodiments, the TCR comprises CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 37 or 42, and / or CDR3β containing or consisting of the amino acid sequence of SEQ ID NO: 47 or 53.

[0202] In some embodiments, the TCR comprises CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 37, and / or CDR3β containing or consisting of the amino acid sequence of SEQ ID NO: 47.

[0203] In some embodiments, the TCR comprises CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 42, and / or CDR3β containing or consisting of the amino acid sequence of SEQ ID NO: 53.

[0204] In some embodiments, the TCR is (i) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 35, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 36, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 40, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 41, or a variant thereof having up to three amino acid substitutions, additions, or deletions. The present invention further comprises α, and / or (i)(a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 45, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 46, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii)(a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 51, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 52, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0205] In some embodiments, the TCR includes (i) the amino acid sequence of SEQ ID NO: 38, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 43, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or an α-chain variable domestically modified The β-chain variable domain comprises (i) the amino acid sequence of SEQ ID NO: 48, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 54, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0206] In some embodiments, the TCR includes or comprises an α-chain comprising (i) the amino acid sequence of SEQ ID NO: 39, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 44, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or (i) the amino acid sequence of SEQ ID NO: 49 or 50, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 55 or 56, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or a β-chain comprising the same.

[0207] In one embodiment, the present invention comprises (i) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 35, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 36, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 37, or a variant thereof having up to three amino acid substitutions, additions, or deletions; or (ii) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 40, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 41, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 42, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or The present invention provides a TCR comprising (i) (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 45, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 46, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 47, or a variant thereof having up to three amino acid substitutions, additions, or deletions; or (ii) (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 51, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 52, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 53, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0208] In one embodiment, the present invention provides a TCR comprising (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 35, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 36, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 37, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or CDR1β comprising (a) CDR1β comprising the amino acid sequence of SEQ ID NO: 45, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising the amino acid sequence of SEQ ID NO: 46, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising the amino acid sequence of SEQ ID NO: 47, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0209] In one embodiment, the present invention provides a TCR comprising (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 40, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 41, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 42, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 51, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 52, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 53, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0210] In some embodiments, the CDR variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR variant has up to three amino acid substitutions. In some embodiments, the CDR variant has up to two amino acid substitutions. In some embodiments, the CDR variant has up to one amino acid substitution.

[0211] In some embodiments, the TCR is (i) (a) CDR1α containing or consisting of the amino acid sequence of SEQ ID NO: 35, (b) CDR2α containing or consisting of the amino acid sequence of SEQ ID NO: 36, (c) CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 37, or (ii) (a) CDR1α containing or consisting of the amino acid sequence of SEQ ID NO: 40, (b) CDR2α containing or consisting of the amino acid sequence of SEQ ID NO: 41, (c) CDR containing or consisting of the amino acid sequence of SEQ ID NO: 42 3α, and / or (i)(a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 45, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 46, (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 47, or (ii)(a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 51, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 52, (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 53.

[0212] In some embodiments, the TCR comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 35, (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 36, (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 37, and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 45, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 46, and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 47.

[0213] In some embodiments, the TCR comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 40, (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 41, (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 42, and / or (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 51, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 52, and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 53.

[0214] In one embodiment, the present invention relates to an α-chain variable domain comprising (i) the amino acid sequence of SEQ ID NO: 38, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 43, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or The present invention provides a TCR comprising a β-chain variable domain, which comprises (i) the amino acid sequence of SEQ ID NO: 48, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 54, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0215] In one embodiment, the present invention relates to an α-chain variable domain comprising or comprising the amino acid sequence of SEQ ID NO: 38, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereof, wherein the α-chain variable domain comprises or comprises the amino acid sequence of SEQ ID NO: 37, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and comprises CDR3α. (ii) an α-chain variable domain, or an α-chain variable domain comprising or consisting thereof, which comprises the amino acid sequence of SEQ ID NO: 43, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises or consists thereof, which comprises the amino acid sequence of SEQ ID NO: 42, or a variant thereof having up to three amino acid substitutions, additions, or deletions. An α-chain variable domain containing DR3α, and / or (i) a β-chain variable domain comprising or consisting thereof, which comprises the amino acid sequence of SEQ ID NO: 48, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises the amino acid sequence of SEQ ID NO: 47, or a variant thereof having up to three amino acid substitutions, additions, or deletions. (ii) a β-chain variable domain comprising CDR3β, or (ii) the amino acid sequence of SEQ ID NO: 54, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises the amino acid sequence of SEQ ID NO: 53, or a variant thereof having up to three amino acid substitutions, additions, or deletions.The present invention provides a TCR comprising a β-chain variable domain containing CDR3β, which is either contained therein or composed of CDR3β.

[0216] In one embodiment, the present invention relates to an α-chain variable domain comprising or comprising (i) the amino acid sequence of SEQ ID NO: 38, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises or comprising (a) the amino acid sequence of SEQ ID NO: 35, or a variant thereof having up to three amino acid substitutions, additions, or deletions. (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 1α, (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 37, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of SEQ ID NO: 43, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, less An α-chain variable domain comprising or consisting of a variant having at least 97%, at least 98%, or at least 99% sequence identity, wherein the α-chain variable domain comprises (a) CDR1α comprising or consisting of the amino acid sequence of SEQ ID NO: 40, or a variant having up to 3 amino acid substitutions, additions, or deletions thereof; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 41, or a variant having up to 3 amino acid substitutions, additions, or deletions thereof; (c) the amino acid sequence of SEQ ID NO: 42, or up to 3 amino acid substitutions, additions, or deletions thereof. CDR3α comprising an α-chain variable domain comprising (i) the amino acid sequence of SEQ ID NO: 48, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises (a) the amino acid sequence of SEQ ID NO: 45,(b) CDR1β containing or comprising the amino acid sequence of SEQ ID NO: 46, or the amino acid sequence of SEQ ID NO: 47, or the amino acid sequence of SEQ ID NO: 47, or the amino acid sequence of SEQ ID NO: 47, or the amino acid sequence of SEQ ID NO: 3β containing or comprising the amino acid sequence of SEQ ID NO: 54, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, A TCR is provided that includes a β-chain variable domain comprising or consisting of a variant having at least 98% or at least 99% sequence identity, wherein the β-chain variable domain comprises: (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 51 or a variant having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 52 or a variant having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 53 or a variant having up to three amino acid substitutions, additions, or deletions.

[0217] In some embodiments, the TCR includes an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 38, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 48, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0218] In some embodiments, the TCR includes an α-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 54, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0219] In one embodiment, the present invention relates to an α-chain comprising (i) the amino acid sequence of SEQ ID NO: 39, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 44, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or (i) sequence (ii) A TCR is provided which includes or comprises a β-chain comprising the amino acid sequence of number 49 or 50, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of number 55 or 56, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0220] In one embodiment, the present invention relates to an α-chain comprising (i) the amino acid sequence of SEQ ID NO: 39, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises CDR3α comprising the amino acid sequence of SEQ ID NO: 35, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) an α chain comprising or consisting of the amino acid sequence of SEQ ID NO: 44, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α chain comprises CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 42, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or (i) sequence A β-chain comprising or consisting of the amino acid sequence of number 49 or 50, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 47, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) sequence number The present invention provides a TCR comprising a β-chain containing or comprising the amino acid sequence of sequence number 55 or 56, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain contains CDR3β, which contains or comprises the amino acid sequence of sequence number 53, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0221] In one embodiment, the present invention relates to an α-chain comprising (i) the amino acid sequence of SEQ ID NO: 39, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises (a) the amino acid sequence of SEQ ID NO: 35, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (b) SEQ ID NO: 3 (c) CDR2α containing or comprising the amino acid sequence of 6, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions; (c) an α-chain containing the amino acid sequence of SEQ ID NO: 37, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions, or CDR3α comprising or comprising the amino acid sequence of SEQ ID NO: 44, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% thereof , or an α-chain comprising or consisting thereof, wherein the α-chain comprises (a) CDR1α comprising or consisting thereof, the amino acid sequence of SEQ ID NO: 40, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting thereof, the amino acid sequence of SEQ ID NO: 41, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions; and (c) CDR2α comprising the amino acid sequence of SEQ ID NO: 42, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions. An α-chain comprising (i) the amino acid sequence of SEQ ID NO: 49 or 50, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises (a) the amino acid sequence of SEQ ID NO: 45, or a variant thereof having up to three amino acid substitutions, additions, or deletions.(b) CDR1β containing or consisting of (a) the amino acid sequence of SEQ ID NO: 46, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR2β containing or consisting of (c) the amino acid sequence of SEQ ID NO: 47, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR3β containing or consisting of (ii) the amino acid sequence of SEQ ID NO: 55 or 56, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, Alternatively, the present invention provides a TCR comprising a β-chain containing or comprising such variants having at least 99% sequence identity, wherein the β-chain comprises (a) CDR1β containing or comprising the amino acid sequence of SEQ ID NO: 51 or a variant thereof having up to three amino acid substitutions, additions, or deletions, (b) CDR2β containing or comprising the amino acid sequence of SEQ ID NO: 52 or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (c) CDR3β containing or comprising the amino acid sequence of SEQ ID NO: 53 or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0222] In some embodiments, the TCR provides an α-chain comprising the amino acid sequence of SEQ ID NO: 39, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain comprising the amino acid sequence of SEQ ID NO: 49 or 50, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0223] In some embodiments, the TCR includes an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 44, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or a β-chain comprising the amino acid sequence of SEQ ID NO: 55 or 56, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0224] Cathepsin G (CTSG) specific TCR In one embodiment, the present invention provides a TCR that binds to a CTSG peptide when presented by an MHC.

[0225] CTSG Cathepsin G (CTSG) is a protein encoded by the CTSG gene.

[0226] Cathepsin G is expressed in the early stages of myeloid differentiation and is involved in the degradation of extracellular matrix components, cleavage of inflammatory mediators, immune responses, and antigen presentation. Today, CTSG is considered a potential leukemia-associated antigen because it is abnormally overexpressed in leukemic blasts and is characterized by an immature phenotype. In fact, CTSG peptides are processed naturally and presented on the surface of leukemia cells, demonstrating that this antigen is an ideal target for adoptive immunotherapy (see Papadopoulos, KP et al., 1997, The Journal of the American Society of Hematology, 90(12), pp.4938-4946).

[0227] The CTSG protein may have the amino acid sequence described in UniProt entry P08311. An exemplary CTSG protein has the amino acid sequence shown in SEQ ID NO: 57 below.

[0228] [Table 11]

[0229] CTSG peptide As used herein, the term "CTSG peptide" may refer to a peptide comprising an amino acid sequence derived from the CTSG protein. Preferably, a CTSG peptide may comprise at least five, at least six, at least seven, at least eight, or at least nine consecutive amino acid residues of the CTSG protein amino acid sequence (or a variant thereof). In some embodiments, the CTSG peptide comprises nine consecutive amino acid residues of the CTSG protein amino acid sequence (or a variant thereof).

[0230] In one embodiment, the present invention provides a CTSG peptide comprising or consisting of an amino acid sequence selected from SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, and SEQ ID NO: 95, and variants thereof having up to three amino acid substitutions, additions, or deletions, respectively. Preferably, the CTSG peptide is an isolated peptide.

[0231] In some embodiments, the CTSG peptide comprises or consists of the amino acid sequences of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or variants thereof (or fragments thereof) having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequences of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or variants thereof (or fragments thereof) having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequences of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or variants thereof (or fragments thereof) having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequences of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or variants thereof (or fragments thereof) having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or variants thereof (or fragments thereof) having up to one amino acid substitution.

[0232] In some embodiments, the CTSG peptide consists of fewer than 14, 13, 12, 11, or 10 amino acid residues. In some embodiments, the CTSG peptide consists of about 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more amino acid residues. In some embodiments, the CTSG peptide has a length of about 9-14 amino acid residues, about 9-13 amino acid residues, about 9-12 amino acid residues, about 9-11 amino acid residues, or about 9-10 amino acid residues. In some embodiments, the CTSG peptide is a necaper fragment.

[0233] In some embodiments, the CTSG peptide consists of a 9-mer fragment of the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or a variant (or fragment thereof) having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide consists of a 9-mer fragment of the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or a variant (or fragment thereof) having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide consists of a 9-mer fragment of the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or a variant (or fragment thereof) having up to three amino acid substitutions. In some embodiments, the CTSG peptide consists of a 9-mer fragment of the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or a variant (or fragment thereof) having up to two amino acid substitutions. In some embodiments, the CTSG peptide consists of a nnomer fragment of the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 535, SEQ ID NO: 81, SEQ ID NO: 95, or a variant thereof (or a fragment thereof) having up to one amino acid substitution.

[0234] Example 1 of CTSG peptide and CTSG-specific TCR sequence An exemplary CTSG peptide has the amino acid sequence shown in SEQ ID NO: 58 below. GIVSYGKSSGVPPEV Exemplary CTSG peptide 1 (SEQ ID NO: 58)

[0235] In one embodiment, the present invention provides a CTSG peptide comprising or comprising the amino acid sequence of SEQ ID NO: 58, or a variant thereof (or a fragment thereof) having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 58, or a variant thereof (or a fragment thereof) having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 58, or a variant thereof (or a fragment thereof) having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 58, or a variant thereof (or a fragment thereof) having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 58, or a variant thereof (or a fragment thereof) having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 58, or a variant thereof (or a fragment thereof) having up to one amino acid substitution. In some embodiments, the CTSG peptide comprises or consists of the amino acid sequence (or a fragment thereof) of SEQ ID NO: 58.

[0236] In one embodiment, the present invention provides a CTSG peptide comprising or comprising a neptamer fragment of the amino acid sequence of SEQ ID NO: 58, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 58, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 58, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 58, or a variant thereof having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 58, or a variant thereof having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 58, or a variant thereof having up to one amino acid substitution. In some embodiments, the CTSG peptide comprises or consists of a 9-mer fragment of the amino acid sequence of SEQ ID NO: 58.

[0237] CTSG peptides may be immunogenic peptides. CTSG peptides can bind to any suitable MHC. Preferably, CTSG peptides bind to MHC encoded by an HLA class I allele or an HLA class II allele. Preferably, CTSG peptides bind to MHC encoded by an HLA-A allele. Preferably, CTSG peptides bind to MHC encoded by an HLA-A allele. * It binds to the MHC encoded by the O2 allele. Preferably, the CTSG peptide is HLA-A * It binds to the MHC encoded by 0201.

[0238] The inventors have determined the amino acid sequence of the CTSG-specific TCR, which includes the CDR region and is involved in the binding specificity to the CTSG peptide.

[0239] The CTSG-specific TCR can be constrained to any suitable MHC (e.g., one or more HLA alleles). Preferably, the CTSG-specific TCR is constrained to an HLA class I or HLA class II allele. Preferably, the CTSG-specific TCR is constrained to an HLA-A allele. Preferably, the CTSG-specific TCR is constrained to an HLA-A allele. * It is constrained to the 02 allele. Preferably, the CTSG-specific TCR is HLA-A * He was detained at 0201.

[0240] Exemplary CTSG-specific TCR amino acid sequences are shown in the table below.

[0241] [Table 12-1]

[0242] [Table 12-2]

[0243] In one embodiment, the present invention provides a TCR comprising (i) the amino acid sequence of SEQ ID NO: 61, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of SEQ ID NO: 66, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or a TCR comprising (i) the amino acid sequence of SEQ ID NO: 71, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of SEQ ID NO: 77, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a TCR comprising (i) the amino acid sequence of SEQ ID NO: 71, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0244] In some embodiments, the CDR3 variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR3 variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR3 variant has up to three amino acid substitutions. In some embodiments, the CDR3 variant has up to two amino acid substitutions. In some embodiments, the CDR3 variant has up to one amino acid substitution.

[0245] In some embodiments, the TCR comprises CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 61 or 66, and / or CDR3β containing or consisting of the amino acid sequence of SEQ ID NO: 71 or 77.

[0246] In some embodiments, the TCR is (i) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 59, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 60, or a variant thereof having up to three amino acid substitutions, additions, or deletions; or (ii) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 64, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 65, or a variant thereof having up to three amino acid substitutions, additions, or deletions. , and / or (i) CDR1β comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 69, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 70, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR1β comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 75, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 76, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0247] In some embodiments, the TCR includes (i) the amino acid sequence of SEQ ID NO: 62, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 67, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or an α-chain variable domedium comprising the same. The β-chain variable domain comprises (i) the amino acid sequence of SEQ ID NO: 72, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 78, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0248] In some embodiments, the TCR includes or comprises an α-chain comprising (i) the amino acid sequence of SEQ ID NO: 63, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 68, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or (i) the amino acid sequence of SEQ ID NO: 73 or 74, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 79 or 80, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or a β-chain comprising the same.

[0249] In one embodiment, the present invention comprises (i) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 59, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 60, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 61, or a variant thereof having up to three amino acid substitutions, additions, or deletions; or (ii) CDR1α comprising or consisting of (a) the amino acid sequence of SEQ ID NO: 64, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 65, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 66, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and / or The present invention provides a TCR comprising (i) (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 69, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 70, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 71, or a variant thereof having up to three amino acid substitutions, additions, or deletions; or (ii) (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 75, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 76, or a variant thereof having up to three amino acid substitutions, additions, or deletions; or (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 77, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0250] In some embodiments, the CDR variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR variant has up to one amino acid substitution, addition, or deletion. In some embodiments, the CDR variant has up to three amino acid substitutions. In some embodiments, the CDR variant has up to two amino acid substitutions. In some embodiments, the CDR variant has up to one amino acid substitution.

[0251] In some embodiments, the TCR is (i) (a) CDR1α containing or consisting of the amino acid sequence of SEQ ID NO: 59, (b) CDR2α containing or consisting of the amino acid sequence of SEQ ID NO: 60, (c) CDR3α containing or consisting of the amino acid sequence of SEQ ID NO: 61, or (ii) (a) CDR1α containing or consisting of the amino acid sequence of SEQ ID NO: 64, (b) CDR2α containing or consisting of the amino acid sequence of SEQ ID NO: 65, or (c) CDR2α containing or consisting of the amino acid sequence of SEQ ID NO: 66 R3α and / or (i)(a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 69, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 70, (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 71, or (ii)(a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 75, (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 76, or (c) CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 77.

[0252] In one embodiment, the present invention comprises or comprises an α-chain variable domain including (i) the amino acid sequence of SEQ ID NO: 62, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 67, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / Alternatively, the present invention provides a TCR comprising (i) the amino acid sequence of SEQ ID NO: 72, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 78, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or a β-chain variable domain comprising such a domain.

[0253] In one embodiment, the present invention relates to an α-chain variable domain comprising or comprising the amino acid sequence of SEQ ID NO: 62, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises or comprises the amino acid sequence of SEQ ID NO: 61, or a variant thereof having up to three amino acid substitutions, additions, or deletions. (ii) an α-chain variable domain, or an α-chain variable domain comprising or consisting thereof, which comprises the amino acid sequence of SEQ ID NO: 67, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises or consists thereof, which comprises the amino acid sequence of SEQ ID NO: 66, or a variant thereof having up to three amino acid substitutions, additions, or deletions. An α-chain variable domain containing DR3α, and / or (i) a β-chain variable domain comprising or consisting thereof, which comprises the amino acid sequence of SEQ ID NO: 72, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises the amino acid sequence of SEQ ID NO: 71, or a variant thereof having up to three amino acid substitutions, additions, or deletions. (ii) a β-chain variable domain comprising CDR3β, or (ii) the amino acid sequence of SEQ ID NO: 78, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises the amino acid sequence of SEQ ID NO: 77, or a variant thereof having up to three amino acid substitutions, additions, or deletions.The present invention provides a TCR comprising a β-chain variable domain containing CDR3β, which is either contained therein or composed of CDR3β.

[0254] In one embodiment, the present invention relates to an α-chain variable domain comprising or comprising (i) the amino acid sequence of SEQ ID NO: 62, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain variable domain comprises or comprises (a) the amino acid sequence of SEQ ID NO: 59, or a variant thereof having up to three amino acid substitutions, additions, or deletions. (b) CDR2α comprising or consisting of the amino acid sequence of SEQ ID NO: 1α, (c) CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 61, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of SEQ ID NO: 67, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, less An α-chain variable domain comprising or consisting of a variant having at least 97%, at least 98%, or at least 99% sequence identity, wherein the α-chain variable domain comprises (a) the amino acid sequence of SEQ ID NO: 64, or a variant having up to 3 amino acid substitutions, additions, or deletions, or CDR1α; (b) the amino acid sequence of SEQ ID NO: 65, or a variant having up to 3 amino acid substitutions, additions, or deletions, or CDR2α; (c) the amino acid sequence of SEQ ID NO: 66, or up to 3 amino acid substitutions, additions, or deletions. CDR3α comprising an α-chain variable domain comprising (i) the amino acid sequence of SEQ ID NO: 72, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain variable domain comprises (a) the amino acid sequence of SEQ ID NO: 69,(b) CDR1β containing or comprising the amino acid sequence of SEQ ID NO: 70 or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR2β containing or comprising the amino acid sequence of SEQ ID NO: 71 or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR3β containing or comprising the amino acid sequence of SEQ ID NO: 78, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, less A TCR is provided that includes a β-chain variable domain comprising or consisting of a variant having at least 98% or 99% sequence identity, wherein the β-chain variable domain comprises: (a) CDR1β comprising or consisting of the amino acid sequence of SEQ ID NO: 75, or a variant having up to three amino acid substitutions, additions, or deletions; (b) CDR2β comprising or consisting of the amino acid sequence of SEQ ID NO: 76, or a variant having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β comprising the amino acid sequence of SEQ ID NO: 77, or a variant having up to three amino acid substitutions, additions, or deletions.

[0255] In one embodiment, the present invention relates to an α-chain comprising or consisting of (i) the amino acid sequence of SEQ ID NO: 63, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of SEQ ID NO: 68, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, and / or (i) sequence (ii) A TCR is provided which includes or comprises a β-chain comprising the amino acid sequence of number 73 or 74, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, or (ii) the amino acid sequence of sequence number 79 or 80, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto.

[0256] In one embodiment, the present invention relates to an α-chain comprising or consisting of the amino acid sequence of SEQ ID NO: 63, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 61, or a variant thereof having up to three amino acid substitutions, additions, or deletions, (ii) an α chain comprising or consisting of the amino acid sequence of SEQ ID NO: 68, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α chain comprises CDR3α comprising or consisting of the amino acid sequence of SEQ ID NO: 66, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or (i) sequence (ii) sequence, a β-chain comprising or consisting of the amino acid sequence of number 73 or 74, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises CDR3β comprising or consisting of the amino acid sequence of SEQ ID NO: 71, or a variant thereof having up to three amino acid substitutions, additions, or deletions. The present invention provides a TCR comprising a β-chain containing or comprising the amino acid sequence of sequence number 79 or 80, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises a β-chain containing CDR3β, which contains or comprises the amino acid sequence of sequence number 77, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0257] In one embodiment, the present invention relates to an α-chain comprising (i) the amino acid sequence of SEQ ID NO: 63, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the α-chain comprises (a) the amino acid sequence of SEQ ID NO: 59, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (b) SEQ ID NO: 6 (c) CDR2α containing or consisting of the amino acid sequence of 0, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions, (c) an α chain containing the amino acid sequence of SEQ ID NO: 61, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions, or consisting of or consisting of CDR3α, or (ii) the amino acid sequence of SEQ ID NO: 68, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% thereof , or an α chain comprising or consisting thereof, wherein the α chain comprises (a) CDR1α comprising or consisting thereof, the amino acid sequence of SEQ ID NO: 64, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions; (b) CDR2α comprising or consisting thereof, the amino acid sequence of SEQ ID NO: 65, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions; (c) CDR2α comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof having up to 3 amino acid substitutions, additions, or deletions. CDR3α comprising or therein, an α-chain and / or (i) the amino acid sequence of SEQ ID NO: 73 or 74, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto, wherein the β-chain comprises (a) the amino acid sequence of SEQ ID NO: 69, or a variant thereof having up to three amino acid substitutions, additions, or deletions.(b) CDR1β containing or consisting of (c) the amino acid sequence of SEQ ID NO: 70, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR2β containing or consisting of (c) the amino acid sequence of SEQ ID NO: 71, or a variant thereof having up to three amino acid substitutions, additions, or deletions, or CDR3β containing or consisting of (ii) the amino acid sequence of SEQ ID NO: 79 or 80, or at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, Alternatively, the present invention provides a TCR comprising a β-chain containing or comprising variants thereof having at least 99% sequence identity, wherein the β-chain comprises (a) CDR1β containing or comprising the amino acid sequence of SEQ ID NO: 75, or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) CDR2β containing or comprising the amino acid sequence of SEQ ID NO: 76, or a variant thereof having up to three amino acid substitutions, additions, or deletions; and (c) CDR3β containing or comprising the amino acid sequence of SEQ ID NO: 77, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0258] Example 2 of CTSG peptides and CTSG-specific TCR sequences Another exemplary CTSG peptide has the amino acid sequence shown in Sequence ID No. 81 below. SGVPPEVFTRVSSFL Example: CTSG Peptide 2 (SEQ ID NO: 81)

[0259] The inventors have determined that residues 2, 3, 4, and 6 of SEQ ID NO: 81 are not essential for MHC binding and / or TCR recognition. Accordingly, in one embodiment, the present invention provides a CTSG peptide comprising a conservative substitution at one or more of positions 2, 3, 4, and / or 6 of SEQ ID NO: 81.

[0260] The CTSG peptide may have the amino acid sequence shown in SEQ ID NO: 535 below. SXXXPXVFTRVSSFL, where X is any amino acid (SEQ ID NO: 535)

[0261] In one embodiment, the present invention provides a CTSG peptide comprising or comprising the amino acid sequence of SEQ ID NO: 535, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 535, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 535, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 535, or a variant thereof having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 535, or a variant thereof having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 535, or a variant thereof having up to one amino acid substitution. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 535. Preferably, the CTSG peptide is HLA-C * It binds to the MHC encoded by the 07 allele. Preferably, the CTSG peptide binds to HLA-C * It binds to MHC encoded by 0702. Preferably, the CTSG peptide binds to HLA-A * It binds to MHC encoded by 24 alleles. Preferably, the CTSG peptide is HLA-A * It binds to MHC encoded by 2402. Preferably, the CTSG peptide binds to HLA-C * MHC and HLA-A encoded by allele 07 * It binds to MHC encoded by 24 alleles. Preferably, the CTSG peptide binds to HLA-C * MHC and HLA-A coded by 0702* It binds to the MHC encoded by 2402.

[0262] In one embodiment, the present invention provides a CTSG peptide comprising or comprising the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to one amino acid substitution. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 81.

[0263] In one embodiment, the present invention provides a CTSG peptide comprising or comprising a neptamer fragment of the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises a neptamer fragment of the amino acid sequence of SEQ ID NO: 81, or a variant thereof having up to one amino acid substitution. In some embodiments, the CTSG peptide comprises or consists of a 9-mer fragment of the amino acid sequence of SEQ ID NO: 81. Preferably, the CTSG peptide is HLA-C * It binds to the MHC encoded by the 07 allele. Preferably, the CTSG peptide binds to HLA-C * It binds to MHC encoded by 0702. Preferably, the CTSG peptide binds to HLA-A * It binds to MHC encoded by 24 alleles. Preferably, the CTSG peptide is HLA-A * It binds to MHC encoded by 2402. Preferably, the CTSG peptide binds to HLA-C * MHC and HLA-A encoded by allele 07 * It binds to MHC encoded by 24 alleles. Preferably, the CTSG peptide binds to HLA-C * MHC and HLA-A coded by 0702 * It binds to the MHC encoded by 2402.

[0264] An exemplary fragment has the amino acid sequence shown below in Sequence ID No. 82. VFTRVSSFL Example: CTSG peptide 2-fragment 1 (SEQ ID NO: 82)

[0265] In one embodiment, the present invention provides a CTSG peptide comprising or comprising the amino acid sequence of SEQ ID NO: 82, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 82, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 82, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 82, or a variant thereof having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 82, or a variant thereof having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 82, or a variant thereof having up to one amino acid substitution. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 82. Preferably, the CTSG peptide is HLA-A * It binds to MHC encoded by 24 alleles. Preferably, the CTSG peptide is HLA-A * It binds to the MHC encoded by 2402.

[0266] Another exemplary fragment has the amino acid sequence shown below in Sequence ID No. 83. VPPEVFTRVSSFL Example: CTSG peptide 2-fragment 2 (SEQ ID NO: 83)

[0267] In one embodiment, the present invention provides a CTSG peptide comprising or comprising the amino acid sequence of SEQ ID NO: 83, or a variant thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 83, or a variant thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 83, or a variant thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 83, or a variant thereof having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 83, or a variant thereof having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 83, or a variant thereof having up to one amino acid substitution. In some embodiments, the CTSG peptide comprises or comprises the amino acid sequence of SEQ ID NO: 83. Preferably, the CTSG peptide is HLA-C * It binds to the MHC encoded by the 07 allele. Preferably, the CTSG peptide binds to HLA-C * It binds to the MHC encoded by 0702.

[0268] Other exemplary fragments have the amino acid sequences of SEQ ID NO: 420, SEQ ID NO: 421, and SEQ ID NO: 422 shown below. EVFTRVSSF Example: CTSG Peptide 2-Fragment 3 (SEQ ID NO: 420) EVFTRVSSFL Example: CTSG peptide 2-fragment 4 (SEQ ID NO: 421) VPPEVFTRVSSF Example: CTSG peptide 2-fragment 5 (SEQ ID NO: 422)

[0269] In one embodiment, the present invention provides a CTSG peptide comprising or comprising any of the amino acid sequences of SEQ ID NOs. 420 to 422, or variants thereof having up to three amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises any of the amino acid sequences of SEQ ID NOs. 420 to 422, or variants thereof having up to two amino acid substitutions, additions, or deletions. In some embodiments, the CTSG peptide comprises or comprises any of the amino acid sequences of SEQ ID NOs. 420 to 422, or variants thereof having up to one amino acid substitution, addition, or deletion. In some embodiments, the CTSG peptide comprises or comprises any of the amino acid sequences of SEQ ID NOs. 420 to 422, or variants thereof having up to three amino acid substitutions. In some embodiments, the CTSG peptide comprises or comprises any of the amino acid sequences of SEQ ID NOs. 420 to 422, or variants thereof having up to two amino acid substitutions. In some embodiments, the CTSG peptide comprises or consists of any of the amino acid sequences of SEQ ID NOs. 420 to 422, or variants thereof having up to one amino acid substitution.

[0270] CTSG peptides may be immunogenic peptides. CTSG peptides can bind to any suitable MHC. Preferably, CTSG peptides bind to MHC encoded by an HLA class I allele or an HLA class II allele. Preferably, CTSG peptides bind to MHC encoded by an HLA-A allele or an HLA-C allele. Preferably, CTSG peptides bind to MHC encoded by an HLA-A allele or an HLA-C allele. * 24 alleles or HLA-C * It binds to the MHC encoded by the 07 allele. Preferably, the CTSG peptide is HLA-A * 2402 or HLA-C *It binds to the MHC encoded by 0702. Preferably, the CTSG peptide binds to the MHC encoded by the HLA-A allele and the HLA-C allele. Preferably, the CTSG peptide binds to the MHC encoded by the HLA-A allele. * MHC and HLA-C encoded by 24 alleles * It binds to the MHC encoded by the 07 allele. Preferably, the CTSG peptide is HLA-A * MHC and HLA-C coded by 2402 * It binds to the MHC encoded by 0702.

[0271] The inventors have determined the amino acid sequence of the CTSG-specific TCR, which includes the CDR region and is involved in the binding specificity to the CTSG peptide.

[0272] The CTSG-specific TCR can be constrained to any suitable MHC (e.g., one or more HLA alleles). Preferably, the CTSG-specific TCR is constrained to an HLA class I allele or an HLA class II allele. Preferably, the CTSG-specific TCR is constrained to an HLA-A allele or an HLA-C allele. Preferably, the CTSG-specific TCR is constrained to an HLA-A allele. * 24 alleles or HLA-C * It is bound to the 07 allele. Preferably, the CTSG-specific TCR is HLA-A * 2402 or HLA-C * It is bound to 0702. Preferably, the CTSG-specific TCR is bound to the HLA-A allele and the HLA-C allele. Preferably, the CTSG-specific TCR is bound to the HLA-A allele * 24 alleles and HLA-C * It is bound to the 07 allele. Preferably, the CTSG-specific TCR is HLA-A * 2402 and HLA-C * He was detained at 0702.

[0273] Exemplary CTSG-specific TCR amino acid sequences are shown in the table below.

[0274] [Table 13]

[0275] As used herein, the terms “TCR P56,” “P56 TCR,” “P56αβ,” “P56 TCRαβ,” and “CTSG TCR” may be used interchangeably and, unless otherwise specified, refer to TCRs comprising an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 87 or a variant thereof, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 92 or a variant thereof.

[0276] In one embodiment, the present invention provides a TCR comprising CDR3α comprising or having the amino acid sequence of SEQ ID NO: 86, or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or CDR3β comprising the amino acid sequence of SEQ ID NO: 91, or a variant thereof having up to three amino acid substitutions, additions, or deletions.

[0277] In some embodiments, the CDR3 variant has up to two amino acid substitutions, additions, or deletions. In some embodiments, the CDR3 variant ha...

Claims

1. A T cell receptor (TCR) that binds to an immunogenic peptide when presented by the major histocompatibility complex (MHC), (1) The immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVQASNDYKLSF (SEQ ID NO: 86), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CassvGrateaff (SEQ ID NO: 91), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (2) The immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CALPSARQLTF (SEQ ID NO: 61) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α containing the amino acid sequence of CAFMSPEGGSEKLVF (SEQ ID NO: 66) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CSVPPAGLGAPEAFF (SEQ ID NO: 71) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSPGTGAYNSPLHF (SEQ ID NO: 77) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (3) The immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CVVNWDNARLMF (SEQ ID NO: 98) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α containing the amino acid sequence of CATRRAKDRDDKIIF (SEQ ID NO: 459) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CASSEEGGSTDQYF (SEQ ID NO: 103) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSLGLAGDYEQYF (SEQ ID NO: 109) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (4) The immunogenic peptide is a mesothelin (MSLN) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAAYNDYKLSF (SEQ ID NO: 293) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSLQGVNTEAFF (SEQ ID NO: 298) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (5) The immunogenic peptide is Dickkopf-related protein 1 (DKK1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAVGAWYNQGGKLIF (SEQ ID NO: 241) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSFGVTGELF (SEQ ID NO: 246) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (6) The immunogenic peptide is a human epidermal growth factor receptor 2 (HER2) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CAEGSRYGGATNKLIF (SEQ ID NO: 8) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence of CASSTFPVETQYF (SEQ ID NO: 13) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (7) The immunogenic peptide is a proteinase 3 (PR3) peptide or a neutrophil elastase (NE) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CADYYGQNFVF (SEQ ID NO: 22), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSFQGYTEAFF (SEQ ID NO: 27), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (8) The immunogenic peptide is a histone-lysine N-methyltransferase EZH2 (EZH2) peptide or a melanoma preferential expression antigen (PRAME) peptide, and the TCR is (i) CDR3α containing the amino acid sequence of CAVSESPTGFQKLVF (SEQ ID NO: 37), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of CAVPHSYNTDKLIF (SEQ ID NO: 42), CDR3α containing a variant of the same having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CASSPRGSNTGELF (SEQ ID NO: 47) or a variant of the same having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CSARPSECQYF (SEQ ID NO: 53) or a variant of the same having up to three amino acid substitutions, additions, or deletions, (9) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAFMKRLTQGGSEKLVF (SEQ ID NO: 119), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSEVYRGHEKLFF (SEQ ID NO: 124), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (10) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAASIRSSGDKLTF (SEQ ID NO: 133), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSKRTGELF (SEQ ID NO: 138), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (11) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CALSETLYNQGGKLIF (SEQ ID NO: 144), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLGTSRSYTDQYF (SEQ ID NO: 149), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (12) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α containing the amino acid sequence of CALTSDYKLSF (SEQ ID NO: 157) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CATSDLFGELF (SEQ ID NO: 162) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASTTGIYEQYF (SEQ ID NO: 168) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (13) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVNSWGKLQF (SEQ ID NO: 176), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASHSGLVGTGELF (SEQ ID NO: 181), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (14) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CASTNFGNEKLTF (SEQ ID NO: 192), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLSYEQYF (SEQ ID NO: 197), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (15) The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR is (i) CDR3α comprising the amino acid sequence of CAVRSHSGNTPLVF (SEQ ID NO: 206) or a variant thereof having up to three amino acid substitutions, additions or deletions, (ii) CDR3α comprising the amino acid sequence of CAENAGGTSYGKLTF (SEQ ID NO: 211) or a variant thereof having up to three amino acid substitutions, additions or deletions, or (iii) the amino acid sequence of CAMRLPIPNNNAGNMLTF (SEQ ID NO: 216) or up to three amino acid substitutions, additions or deletions CDR3α containing a variant having (i) the amino acid sequence of CASSSSPRVGPLYEQYF (SEQ ID NO: 221), or a variant having up to three amino acid substitutions, additions, or deletions, CDR3β containing (ii) the amino acid sequence of CASSDHDIYNEQFF (SEQ ID NO: 227), or a variant having up to three amino acid substitutions, additions, or deletions, CDR3β containing (iii) the amino acid sequence of CASWAEEEEETQYF (SEQ ID NO: 233), or a variant having up to three amino acid substitutions, additions, or deletions, (16) The immunogenic peptide is a mucin-5AC (MUC5AC) peptide, and the TCR comprises CDR3α containing the amino acid sequence CAGGGSGAGSYQLTF (SEQ ID NO: 254) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β containing the amino acid sequence CAISDPTGDNQPQHF (SEQ ID NO: 259) or a variant thereof having up to three amino acid substitutions, additions, or deletions, (17) The immunogenic peptide is a forward gradient 2 (AGR2) peptide, and the TCR comprises CDR3α, which has the amino acid sequence CAVQDYGQNFVF (SEQ ID NO: 267) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which has the amino acid sequence CasspTGSEQYF (SEQ ID NO: 272) or a variant thereof having up to three amino acid substitutions, additions, or deletions. (18) The immunogenic peptide is a c-MET peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAAAAPNNNNDDMRF (SEQ ID NO: 280), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLLAGGSSYEQYF (SEQ ID NO: 285), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (19) The immunogenic peptide is a CTD phosphatase subunit 1 (CTDP1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAMREGTSGTYKYIF (SEQ ID NO: 306), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSNSASRPEQYV (SEQ ID NO: 311), or a variant thereof having up to three amino acid substitutions, additions, or deletions. (20) The immunogenic peptide is a transformed acid-coiled-coil-containing protein 2 (TACC2) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVSGLGGSNYKLTF (SEQ ID NO: 319), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASRSGTGTMDEQFF (SEQ ID NO: 324), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (21) A T cell receptor (TCR) wherein the immunogenic peptide is a prostaglandin F2 receptor negative regulator (PTGFRN) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVNRDDKIIF (SEQ ID NO: 332), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CSARDYKQVLRGSYNSPLHF (SEQ ID NO: 337), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

2. The TCR according to claim 1, wherein the immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVQASNDYKLSF (SEQ ID NO: 86), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSVGRATEAFF (SEQ ID NO: 91), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

3. The TCR according to claim 2, wherein the TCR comprises CDR3α containing the amino acid sequence of CAVQASNDYKLSF (SEQ ID NO: 86) and CDR3β containing the amino acid sequence of CASSVGRATEAFF (SEQ ID NO: 91).

4. The TCR according to claim 2 or 3, wherein the TCR comprises the following CDR sequences: CDR1α-VSGLRG (SEQ ID NO: 84), CDR2α-LYSAGEE (SEQ ID NO: 85), CDR3α-CAVQASNDYKLSF (SEQ ID NO: 86), CDR1β-MNHEY (SEQ ID NO: 89), CDR2β-SMNVEV (SEQ ID NO: 90), and CDR3β-CASSVGRATEAFF (SEQ ID NO: 91), or variants thereof having up to three amino acid substitutions, additions, or deletions.

5. The TCR according to any one of claims 2 to 4, wherein the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 87, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 92, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

6. The TCR according to any one of claims 2 to 5, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 88, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 93 or 94, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

7. The CTSG peptide comprises or consists of the amino acid sequence SXXXXPXVFTRVSSFL (SEQ ID NO: 535), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof, preferably the CTSG peptide comprises or consists of the amino acid sequence SGVPPEVFTRVSSFL (SEQ ID NO: 81), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or a fragment thereof, preferably the CTSG peptide comprises (a) SGV A TCR according to any one of claims 2 to 6, comprising or consisting of: (b) the amino acid sequence of PPEVFTRVSSFL (SEQ ID NO: 81), or a variant thereof having up to three amino acid substitutions, additions, or deletions; (b) the amino acid sequence of VFTRVSSFL (SEQ ID NO: 82), or a variant thereof having up to three amino acid substitutions, additions, or deletions; or (c) the amino acid sequence of VPPEVFTRVSSFL (SEQ ID NO: 83), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

8. The TCR according to claim 1, wherein the immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CALPSARQLTF (SEQ ID NO: 61) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α containing the amino acid sequence of CAFMSPEGGSEKLVF (SEQ ID NO: 66) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CSVPPAGLGAPEAFF (SEQ ID NO: 71) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSPGTGAYNSPLHF (SEQ ID NO: 77) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

9. The TCR according to claim 8, wherein the TCR comprises (i) CDR3α containing the amino acid sequence of CALPSARQLTF (SEQ ID NO: 61), or (ii) CDR3α containing the amino acid sequence of CAFMSPEGGSEKLVF (SEQ ID NO: 66), and (i) CDR3β containing the amino acid sequence of CSVPPAGLGAPEAFF (SEQ ID NO: 71), or (ii) CDR3β containing the amino acid sequence of CASSPGTGAYNSPLHF (SEQ ID NO: 77).

10. The TCR is one of the following CDR sequences: (i) CDR1α-ATGYPS (SEQ ID NO: 59), CDR2α-ATKADDK (SEQ ID NO: 60), and CDR3α-CALPSARQLTF (SEQ ID NO: 61), or (ii) CDR1α-TSENNYY (SEQ ID NO: 64), CDR2α-QEAYKQQN (SEQ ID NO: 65), and CDR3α-CAFMSPEGGSEKLVF (SEQ ID NO: 66), and (i) CDR1β-SQVTM (SEQ ID NO: 6 9) The TCR according to claim 8 or 9, comprising CDR2β-ANQGSEA (SEQ ID NO: 70), CDR3β-CSVPPAGLGAPEAFF (SEQ ID NO: 71), or (ii) CDR1β-SEHNR (SEQ ID NO: 75), CDR2β-FQNEAQ (SEQ ID NO: 76), and CDR3β-CASSPGTGAYNSPLHF (SEQ ID NO: 77), or variants thereof having up to three amino acid substitutions, additions, or deletions.

11. The TCR comprises (i) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 62, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 67, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and (i) SEQ ID NO: 7 A TCR according to any one of claims 8 to 10, comprising: (ii) a β-chain variable domain comprising: (ii) the amino acid sequence of SEQ ID NO: 78, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (ii) a β-chain variable domain comprising: (ii) the amino acid sequence of SEQ ID NO: 78, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

12. The TCR comprises (i) an α chain containing the amino acid sequence of SEQ ID NO: 63, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) an α chain containing the amino acid sequence of SEQ ID NO: 68, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and (i) an α chain containing the amino acid sequence of SEQ ID NO: 73 or 74 A TCR according to any one of claims 8 to 11, comprising (ii) a β-chain comprising an amino acid sequence, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) a β-chain comprising the amino acid sequence of SEQ ID NO: 79 or 80, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

13. The TCR according to any one of claims 8 to 12, wherein the CTSG peptide comprises or consists of a necaper fragment of the amino acid sequence GIVSYGKSSGVPEV (SEQ ID NO: 58), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

14. The TCR according to claim 1, wherein the immunogenic peptide is a cathepsin G (CTSG) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CVVNWDNARLMF (SEQ ID NO: 98) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3α containing the amino acid sequence of CATRRAKDRDDKIIF (SEQ ID NO: 459) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (i) CDR3β containing the amino acid sequence of CASSEEGGSTDQYF (SEQ ID NO: 103) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSLGLAGDYEQYF (SEQ ID NO: 109) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

15. The TCR according to claim 14, wherein the TCR comprises (i) CDR3α containing the amino acid sequence of CVVNWDNARLMF (SEQ ID NO: 98), or (ii) CDR3α containing the amino acid sequence of CATRRAKDRDDKIIF (SEQ ID NO: 459), and (i) CDR3β containing the amino acid sequence of CASSEEGGSDTQYF (SEQ ID NO: 103), or (ii) CDR3β containing the amino acid sequence of CASSLGLAGDYEQYF (SEQ ID NO: 109).

16. The TCR is one of the following CDR sequences: (i) CDR1α-NSASQS (SEQ ID NO: 96), CDR2α-VYSSGN (SEQ ID NO: 97), and CDR3α-CVVNWDNARLMF (SEQ ID NO: 98), or (ii) CDR1α-NSAFQY (SEQ ID NO: 457), CDR2α-TYSSGN (SEQ ID NO: 458), and CDR3α-CATRRAKDRDDKIIF (SEQ ID NO: 459), and (i) CDR1β-SNHLY (SEQ ID NO: 10 1) The TCR according to claim 14 or 15, comprising CDR2β-FYNNEI (SEQ ID NO: 102) and CDR3β-CASSEEGGSTDTQYF (SEQ ID NO: 103), or (ii) CDR1β-SGHNS (SEQ ID NO: 107), CDR2β-FNNNVP (SEQ ID NO: 108) and CDR3β-CASSLGLAGDYEQYF (SEQ ID NO: 109), or variants thereof having up to three amino acid substitutions, additions, or deletions.

17. The TCR comprises (i) an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 99, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 460, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and (i) SEQ ID NO: 10 A TCR according to any one of claims 14 to 16, comprising: (ii) a β-chain variable domain comprising: (ii) the amino acid sequence of SEQ ID NO: 110, or

18. The TCR comprises (i) an α chain containing the amino acid sequence of SEQ ID NO: 100, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) an α chain containing the amino acid sequence of SEQ ID NO: 461, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and (i) an α chain containing the amino acid sequence of SEQ ID NO: 105 or 106 A TCR according to any one of claims 14 to 17, comprising (ii) a β chain comprising an amino acid sequence, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) a β chain comprising the amino acid sequence of SEQ ID NO: 111 or 112, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

19. The TCR according to any one of claims 14 to 18, wherein the CTSG peptide comprises or consists of a necaper fragment of the amino acid sequence TMRSFKLLDQMETPL (SEQ ID NO: 95), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

20. The TCR according to claim 1, wherein the immunogenic peptide is a human epidermal growth factor receptor 2 (HER2) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAEGSRYGGATNKLIF (SEQ ID NO: 8), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSTFPVETQYF (SEQ ID NO: 13), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

21. The TCR according to claim 20, wherein the TCR comprises CDR3α containing the amino acid sequence of CAEGSRYGGATNKLIF (SEQ ID NO: 8) and CDR3β containing the amino acid sequence of CASSTFPVETQYF (SEQ ID NO: 13).

22. The TCR according to claim 20 or 21, wherein the TCR comprises the following CDR sequences: CDR1α-NSASDY (SEQ ID NO: 6), CDR2α-IRSNMDK (SEQ ID NO: 7), CDR3α-CAEGSRYGGATNKLIF (SEQ ID NO: 8), CDR1β-MNHEY (SEQ ID NO: 11), CDR2β-SVGAGI (SEQ ID NO: 12), and CDR3β-CASSTFPVETQYF (SEQ ID NO: 13), or variants thereof having up to three amino acid substitutions, additions, or deletions.

23. The TCR according to any one of claims 20 to 22, wherein the TCR comprises an α-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

24. The TCR according to any one of claims 20 to 23, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 10, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 15 or 16, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

25. The TCR according to any one of claims 20 to 24, wherein the HER2 peptide comprises or consists of the amino acid sequence of KIFGSLAFL (SEQ ID NO: 5), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

26. The TCR according to claim 1, wherein the immunogenic peptide is proteinase 3 (PR3) peptide or neutrophil elastase (NE) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CADYYGQNFVF (SEQ ID NO: 22), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSFQGYTEAFF (SEQ ID NO: 27), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

27. The TCR according to claim 26, wherein the TCR comprises CDR3α containing the amino acid sequence CADYYGQNFVF (SEQ ID NO: 22) and CDR3β containing the amino acid sequence CASSFQGYTEAFF (SEQ ID NO: 27).

28. The TCR according to claim 26 or 27, wherein the TCR comprises the following CDR sequences: CDR1α-VSGNPY (SEQ ID NO: 20), CDR2α-YITGDNLV (SEQ ID NO: 21), CDR3α-CADYYGQNFVF (SEQ ID NO: 22), CDR1β-MDHEN (SEQ ID NO: 25), CDR2β-SYDVKM (SEQ ID NO: 26), and CDR3β-CASSFQGYTEAFF (SEQ ID NO: 27), or variants thereof having up to three amino acid substitutions, additions, or deletions.

29. The TCR according to any one of claims 26 to 28, wherein the TCR comprises an α-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

30. The TCR according to any one of claims 26 to 29, wherein the TCR comprises an α chain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

31. The TCR according to any one of claims 26 to 30, wherein the peptide comprises or consists of the amino acid sequence of VLQELNVTV (SEQ ID NO: 19), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

32. The immunogenic peptide is a histone-lysine N-methyltransferase EZH2 (EZH2) peptide or a melanoma preferential expression antigen (PRAME) peptide, and the TCR is (i) CDR3α containing the amino acid sequence of CAVSESPTGFQKLVF (SEQ ID NO: 37), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of CAVPHSYNTDKLIF (SEQ ID NO: 42), or up to three amino acid substitutions, additions, or deletions. The TCR according to claim 1, comprising (i) CDR3α having a variant thereof having amino acid substitutions, additions, or deletions, and (ii) CDR3β having the amino acid sequence of CASSPRGSNTGELFF (SEQ ID NO: 47) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β having the amino acid sequence of CSARPSECQYF (SEQ ID NO: 53) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

33. The TCR according to claim 32, wherein the TCR comprises (i) CDR3α containing the amino acid sequence of CAVSESPTGFQKLVF (SEQ ID NO: 37), or (ii) CDR3α containing the amino acid sequence of CAVPHSYNTDKLIF (SEQ ID NO: 42), and (i) CDR3β containing the amino acid sequence of CASSPRGSNTGELFF (SEQ ID NO: 47), or (ii) CDR3β containing the amino acid sequence of CSARPSECQYF (SEQ ID NO: 53).

34. The TCR is one of the following CDR sequences: (i) CDR1α-SSVPPPY (SEQ ID NO: 35), CDR2α-YTSAATLV (SEQ ID NO: 36), and CDR3α-CAVSESPTGFQKLVF (SEQ ID NO: 37), or (ii) CDR1α-VGISA (SEQ ID NO: 40), CDR2α-LSSGK (SEQ ID NO: 41), and CDR3α-CAVPHSYNTDKLIF (SEQ ID NO: 42), and (i) CDR1β-SNHLY (Sequence No. The TCR according to claim 32 or 33, comprising (i) CDR2β-FYNNEI (SEQ ID NO: 45), CDR3β-CASSPRGSNTGELFF (SEQ ID NO: 47), or (ii) CDR1β-DFQATT (SEQ ID NO: 51), CDR2β-SNEGSKA (SEQ ID NO: 52), and CDR3β-CSARPSECQYF (SEQ ID NO: 53), or variants thereof having up to three amino acid substitutions, additions, or deletions.

35. The TCR comprises (i) an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 38, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 43, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and (i) SEQ ID NO: 48 A TCR according to any one of claims 32 to 34, comprising: (ii) a β-chain variable domain comprising: (ii) the amino acid sequence of SEQ ID NO: 54, or

36. The TCR comprises (i) an α chain containing the amino acid sequence of SEQ ID NO: 39, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) an α chain containing the amino acid sequence of SEQ ID NO: 44, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and (i) the amino acid sequence of SEQ ID NO: 49 or 50 A TCR according to any one of claims 32 to 35, comprising (ii) a β-chain comprising an amino acid sequence, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, or (ii) a β-chain comprising the amino acid sequence of SEQ ID NO 55 or 56, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

37. The TCR according to any one of claims 32 to 36, wherein the EZH2 peptide comprises or consists of the amino acid sequence of KESRPPRKF (SEQ ID NO: 32) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) a preferred expression antigen (PRAME) peptide for melanoma as presented by MHC, wherein the PRAME peptide comprises or consists of the amino acid sequence of ALYVDSLFFL (SEQ ID NO: 34) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

38. The TCR according to claim 1, wherein the immunogenic peptide is cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence CAFMKRLTQGGSEKLVF (SEQ ID NO: 119), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CASSEVYRGHEKLFF (SEQ ID NO: 124), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

39. The TCR according to claim 38, wherein the TCR comprises CDR3α containing the amino acid sequence CAFMKRLTQGGSEKLVF (SEQ ID NO: 119) and CDR3β containing the amino acid sequence CASSEVYRGHEKLFF (SEQ ID NO: 124).

40. The TCR according to claim 38 or 39, wherein the TCR comprises the following CDR sequences: CDR1α-TSENNYY (SEQ ID NO: 117), CDR2α-QEAYKQQN (SEQ ID NO: 118), CDR3α-CAFMKRLTQGGSEKLVF (SEQ ID NO: 119), CDR1β-SNHLY (SEQ ID NO: 122), CDR2β-FYNNEI (SEQ ID NO: 123), and CDR3β-CASSEVYRGHEKLFF (SEQ ID NO: 124), or variants thereof having up to three amino acid substitutions, additions, or deletions.

41. The TCR according to any one of claims 38 to 40, wherein the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 120, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 125, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

42. The TCR according to any one of claims 38 to 41, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 121, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 126 or 127, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

43. The TCR according to any one of claims 38 to 42, wherein the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of TEYAEEIYQYL (SEQ ID NO: 114) or a variant thereof having up to three amino acid substitutions, additions, or deletions, preferably the CCNA1 peptide comprises or consists of the amino acid sequence of YAEEIYQYL (SEQ ID NO: 115) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

44. The TCR according to claim 1, wherein the immunogenic peptide is cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAASIRSSGDKLTF (SEQ ID NO: 133), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSKRTGELF (SEQ ID NO: 138), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

45. The TCR according to claim 44, wherein the TCR comprises CDR3α containing the amino acid sequence of CAASIRSSGDKLTF (SEQ ID NO: 133) and CDR3β containing the amino acid sequence of CASSKRTGELF (SEQ ID NO: 138).

46. The TCR according to claim 44 or 45, wherein the TCR comprises the following CDR sequences: CDR1α-DSASNY (SEQ ID NO: 131), CDR2α-IRSNVGE (SEQ ID NO: 132), CDR3α-CAASIRSSGDKLTF (SEQ ID NO: 133), CDR1β-MDHEN (SEQ ID NO: 136), CDR2β-SYDVKM (SEQ ID NO: 137), and CDR3β-CASSKRTGELF (SEQ ID NO: 138), or variants thereof having up to three amino acid substitutions, additions, or deletions.

47. The TCR according to any one of claims 44 to 46, wherein the TCR comprises an α-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

48. The TCR according to any one of claims 44 to 47, comprising an α chain containing the amino acid sequence of SEQ ID NO: 135, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 140 or 141, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

49. The TCR according to any one of claims 44 to 48, wherein the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of PETLAAFTGYSLSEI (SEQ ID NO: 128), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

50. The TCR according to claim 1, wherein the immunogenic peptide is cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CALSETLYNQGGKLIF (SEQ ID NO: 144), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLGTSRSYTDQYF (SEQ ID NO: 149), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

51. The TCR according to claim 50, wherein the TCR comprises CDR3α containing the amino acid sequence of CALSETLYNQGGKLIF (SEQ ID NO: 144) and CDR3β containing the amino acid sequence of CASSLGTSRSYTDQYF (SEQ ID NO: 149).

52. The TCR according to claim 50 or 51, wherein the TCR comprises the following CDR sequences: CDR1α-TRDTTYY (SEQ ID NO: 142), CDR2α-RNSFDEQN (SEQ ID NO: 143), CDR3α-CALSETLYNQGGKLIF (SEQ ID NO: 144), CDR1β-SGHTA (SEQ ID NO: 147), CDR2β-FQGNSA (SEQ ID NO: 148), and CDR3β-CASSLGTSRSYTDTQYF (SEQ ID NO: 149), or variants thereof having up to three amino acid substitutions, additions, or deletions.

53. The TCR according to any one of claims 50 to 52, wherein the TCR comprises an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 145, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 150, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

54. The TCR according to any one of claims 50 to 53, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 146, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 151 or 152, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

55. The TCR according to claim 1, wherein the immunogenic peptide is cyclin A1 (CCNA1) peptide, and the TCR comprises (i) CDR3α containing the amino acid sequence of CALTSDYKLSF (SEQ ID NO: 157) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and (ii) CDR3β containing the amino acid sequence of CATSDLFGELF (SEQ ID NO: 162) or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASTTGIYEQYF (SEQ ID NO: 168) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

56. The TCR according to claim 55, wherein the TCR comprises (i) CDR3α containing the amino acid sequence of CALTSDYKLSF (SEQ ID NO: 157), and (ii) CDR3β containing the amino acid sequence of CATSDLFGELF (SEQ ID NO: 162), or (ii) CASTTGIYEQYF (SEQ ID NO: 168).

57. The TCR according to claim 55 or 56, wherein the TCR comprises the following CDR sequences: CDR1α-SSNFYA (SEQ ID NO: 155), CDR2α-MTLNGDE (SEQ ID NO: 156), and CDR3α-CALTSDYKLSF (SEQ ID NO: 157), and (i) CDR1β-KGHDR (SEQ ID NO: 160), CDR2β-SFDVKD (SEQ ID NO: 161), and CDR3β-CATSDLFGELFF (SEQ ID NO: 162), or (ii) CDR1β-SNHLY (SEQ ID NO: 166), CDR2β-FYNNEI (SEQ ID NO: 167), and CDR3β-CASTTGIYEQYF (SEQ ID NO: 168), or variants thereof having up to three amino acid substitutions, additions, or deletions.

58. The TCR according to any one of claims 55 to 57, wherein the TCR comprises (i) an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 158, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (ii) a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 163, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (ii) a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 169, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

59. The TCR according to any one of claims 55 to 58, comprising: (i) an α chain comprising the amino acid sequence of SEQ ID NO: 159, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (ii) a β chain comprising the amino acid sequence of SEQ ID NO: 164 or 165, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (ii) a β chain comprising the amino acid sequence of SEQ ID NO: 170 or 171, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

60. The TCR according to any one of claims 55 to 59, wherein the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of AELSLLEADPFLKYL (SEQ ID NO: 153) or a variant thereof having up to three amino acid substitutions, additions, or deletions, and / or the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of LLEADPFLKYLPSLI (SEQ ID NO: 527) or a variant thereof having up to three amino acid substitutions, additions, or deletions.

61. The TCR according to claim 1, wherein the immunogenic peptide is cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVNSWGKLQF (SEQ ID NO: 176), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASHSGLVGTGELF (SEQ ID NO: 181), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

62. The TCR according to claim 61, wherein the TCR comprises CDR3α containing the amino acid sequence of CAVNSWGKLQF (SEQ ID NO: 176) and CDR3β containing the amino acid sequence of CASHSGLVGTGELF (SEQ ID NO: 181).

63. The TCR according to claim 61 or 62, wherein the TCR comprises the following CDR sequences: CDR1α-TSGFNG (SEQ ID NO: 174), CDR2α-NVLDGL (SEQ ID NO: 175), CDR3α-CAVNSWGKLQF (SEQ ID NO: 176), CDR1β-MNHEY (SEQ ID NO: 179), CDR2β-SMNVEV (SEQ ID NO: 180), and CDR3β-CASHSGLVGTGELF (SEQ ID NO: 181), or variants thereof having up to three amino acid substitutions, additions, or deletions.

64. The TCR according to any one of claims 61 to 63, wherein the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 177, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 182, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

65. The TCR according to any one of claims 61 to 64, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 178, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 183 or 184, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

66. The TCR according to any one of claims 61 to 65, wherein the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of WEGPGLPDFVF (SEQ ID NO: 172), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

67. The TCR according to claim 1, wherein the immunogenic peptide is cyclin A1 (CCNA1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CASTNFGNEKLTF (SEQ ID NO: 192), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLSYEQYF (SEQ ID NO: 197), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

68. The TCR according to claim 67, wherein the TCR comprises CDR3α containing the amino acid sequence of CASTNFGNEKLTF (SEQ ID NO: 192) and CDR3β containing the amino acid sequence of CASSLSYEQYF (SEQ ID NO: 197).

69. The TCR according to claim 67 or 68, wherein the TCR comprises the following CDR sequences: CDR1α-TSESDYY (SEQ ID NO: 190), CDR2α-QEAYKQQN (SEQ ID NO: 191), CDR3α-CASTNFGNEKLTF (SEQ ID NO: 192), CDR1β-PRHDT (SEQ ID NO: 195), CDR2β-FYEKMQ (SEQ ID NO: 196), and CDR3β-CASSLSYEQYF (SEQ ID NO: 197), or variants thereof having up to three amino acid substitutions, additions, or deletions.

70. The TCR according to any one of claims 67 to 69, wherein the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 193, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 198, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

71. The TCR according to any one of claims 67 to 70, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 194, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 199 or 200, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

72. The TCR according to any one of claims 67 to 71, wherein the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of KRQLLKMEHLLLLKVL (SEQ ID NO: 185), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

73. The immunogenic peptide is a cyclin A1 (CCNA1) peptide, and the TCR is a CDR3α comprising (i) the amino acid sequence of CAVRSHSGNTPLVF (SEQ ID NO: 206), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) the amino acid sequence of CAENAGGTSYGKLTF (SEQ ID NO: 211), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (iii) the amino acid sequence of CAMRLPIPNNNAGNMLTF (SEQ ID NO: 216), or a variant thereof having up to three amino acid substitutions, additions, or deletions The TCR according to claim 1, comprising CDR3α containing ant, and CDR3β containing (i) the amino acid sequence of CASSSSPRVGPLYEQYF (SEQ ID NO: 221), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (ii) CDR3β containing the amino acid sequence of CASSDHDIYNEQFF (SEQ ID NO: 227), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or (iii) CDR3β containing the amino acid sequence of CASWAEEAEETQYF (SEQ ID NO: 233), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

74. The TCR according to claim 73, wherein the TCR comprises (i) CDR3α containing the amino acid sequence of CAVRSHSGNTPLVF (SEQ ID NO: 206), (ii) CDR3α containing the amino acid sequence of CAENAGGTSYGKLTF (SEQ ID NO: 211), or (iii) CDR3α containing the amino acid sequence of CAMRLPIPNNNAGNMLTF (SEQ ID NO: 216), and (i) CDR3β containing the amino acid sequence of CASSSSPRVGPLYEQYF (SEQ ID NO: 221), (ii) CDR3β containing the amino acid sequence of CASSDHDIYNEQFF (SEQ ID NO: 227), or (iii) CDR3β containing the amino acid sequence of CASWAEEEAEETQYF (SEQ ID NO: 233).

75. The TCR is one of the following CDR sequences: (i) CDR1α-TSGFYG (SEQ ID NO: 204), CDR2α-NALDGL (SEQ ID NO: 205), and CDR3α-CAVRSHSGNTPLVF (SEQ ID NO: 206), or (ii) CDR1α-NSASDY (SEQ ID NO: 209), CDR2α-IRSNMDK (SEQ ID NO: 210), and CDR3α-CAENAGGTSYGKLTF (SEQ ID NO: 211), or (iii) CDR1α-TSDQSYG (SEQ ID NO: 214), CDR2α-QGSYDEQN (SEQ ID NO: 215), and CDR3α-CAMRRLPIPNNNAGNMLTF (SEQ ID NO: 216), and (i) CDR1β-SEHNR ( The TCR according to claim 73 or 74, comprising (SEQ ID NO: 219), CDR2β-FQNEAQ (SEQ ID NO: 220), and CDR3β-CASSSSPRVGPLYEQYF (SEQ ID NO: 221), or (ii) CDR1β-SEHNR (SEQ ID NO: 225), CDR2β-FQNEAQ (SEQ ID NO: 226), and CDR3β-CASSDHDIYNEQFF (SEQ ID NO: 227), or (iii) CDR1β-SNHLY (SEQ ID NO: 231), CDR2β-FYNNEI (SEQ ID NO: 232), and CDR3β-CASWAEEEEETQYF (SEQ ID NO: 233), or variants thereof having up to three amino acid substitutions, additions, or deletions.

76. The TCR comprises (i) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 207, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (ii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 212, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (iii) an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 217, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and (i) SEQ ID NO: 22 A TCR according to any one of claims 73 to 75, comprising: (ii) a β-chain variable domain comprising: (ii) the amino acid sequence of SEQ ID NO: 228, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (iii) a β-chain variable domain comprising: (ii) the amino acid sequence of SEQ ID NO: 228, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (iii) a β-chain variable domain comprising: (ii) the amino acid sequence of SEQ ID NO: 234, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

77. The TCR comprises (i) an α chain containing the amino acid sequence of SEQ ID NO: 208, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (ii) an α chain containing the amino acid sequence of SEQ ID NO: 213, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; or (iii) an α chain containing the amino acid sequence of SEQ ID NO: 218, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and (i) the amino acid sequence of SEQ ID NO: 223 or 224 A TCR according to any one of claims 73 to 76, comprising (ii) the amino acid sequence of SEQ ID NO: 229 or 230, or a β-chain comprising

78. The TCR according to any one of claims 73 to 77, wherein the CCNA1 peptide comprises or consists of a necaper fragment of the amino acid sequence of PETLAAFTGYSLSEI (SEQ ID NO: 201), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

79. The TCR according to claim 1, wherein the immunogenic peptide is Dickkopf-related protein 1 (DKK1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVGAWYNQGGKLIF (SEQ ID NO: 241), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSFGVTGELF (SEQ ID NO: 246), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

80. The TCR according to claim 79, wherein the TCR comprises CDR3α containing the amino acid sequence CAVGAWYNQGGKLIF (SEQ ID NO: 241) and CDR3β containing the amino acid sequence CASSFGVTGELF (SEQ ID NO: 246).

81. The TCR according to claim 79 or 80, wherein the TCR comprises the following CDR sequences: CDR1α-YGATPY (SEQ ID NO: 239), CDR2α-YFSGDTLV (SEQ ID NO: 240), CDR3α-YFSGDTLV (SEQ ID NO: 241), CDR1β-SGHDY (SEQ ID NO: 244), CDR2β-FNNNNVP (SEQ ID NO: 245), and CDR3β-CASSFGVTGELF (SEQ ID NO: 246), or variants thereof having up to three amino acid substitutions, additions, or deletions.

82. The TCR according to any one of claims 79 to 81, wherein the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 242, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 247, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

83. The TCR according to any one of claims 79 to 82, wherein the TCR comprises an α chain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

84. The TCR according to any one of claims 79 to 83, wherein the DKK1 peptide comprises or consists of the amino acid sequence of ILYPGGNKV (SEQ ID NO: 238), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

85. The TCR according to claim 1, wherein the immunogenic peptide is a mucin-5AC (MUC5AC) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence CAGGGSGAGSYQLTF (SEQ ID NO: 254), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CAISDPTGDNQPQHF (SEQ ID NO: 259), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

86. The TCR according to claim 85, wherein the TCR comprises CDR3α containing the amino acid sequence CAGGGSGAGSYQLTF (SEQ ID NO: 254) and CDR3β containing the amino acid sequence CAISDPTGDNQPQHF (SEQ ID NO: 259).

87. The TCR according to claim 85 or 86, wherein the TCR comprises the following CDR sequences: CDR1α-DRGSQS (SEQ ID NO: 252), CDR2α-IYSNGD (SEQ ID NO: 253), CDR3α-CAGGGSGAGSYQLTF (SEQ ID NO: 254), CDR1β-ENHRY (SEQ ID NO: 257), CDR2β-SYGVKD (SEQ ID NO: 258), and CDR3β-CAISDPTGDNQPQHF (SEQ ID NO: 259), or variants thereof having up to three amino acid substitutions, additions, or deletions.

88. The TCR according to any one of claims 85 to 87, wherein the TCR comprises an α-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

89. The TCR according to any one of claims 85 to 88, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 256, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 261 or 262, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

90. The TCR according to any one of claims 85 to 89, wherein the MUC5AC peptide comprises or consists of the amino acid sequence of FLNDAGACV (SEQ ID NO: 251), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

91. The TCR according to claim 1, wherein the immunogenic peptide is a forward gradient 2 (AGR2) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence CAVQDYGQNFVF (SEQ ID NO: 267), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence CasspTGSEQYF (SEQ ID NO: 272), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

92. The TCR according to claim 91, wherein the TCR comprises CDR3α containing the amino acid sequence CAVQDYGQNFVF (SEQ ID NO: 267) and CDR3β containing the amino acid sequence CASSPTGSEQYF (SEQ ID NO: 272).

93. The TCR according to claim 91 or 92, wherein the TCR comprises the following CDR sequences: CDR1α-VSGLRG (SEQ ID NO: 265), CDR2α-LYSAGEE (SEQ ID NO: 266), CDR3α-CAVQDYGQNFVF (SEQ ID NO: 267), CDR1β-MNHEY (SEQ ID NO: 270), CDR2β-SMNVEV (SEQ ID NO: 271), and CDR3β-CASSPTGSEQYF (SEQ ID NO: 272), or variants thereof having up to three amino acid substitutions, additions, or deletions.

94. The TCR according to any one of claims 91 to 93, wherein the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 268, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 273, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

95. The TCR according to any one of claims 91 to 94, wherein the TCR comprises an α chain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

96. The TCR according to any one of claims 91 to 95, wherein the AGR2 peptide comprises or consists of the amino acid sequence of LLVALSYTL (SEQ ID NO: 264), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

97. The TCR according to claim 1, wherein the immunogenic peptide is a c-MET peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAAAAPNNNNDDMRF (SEQ ID NO: 280), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLLAGGSSYEQYF (SEQ ID NO: 285), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

98. The TCR according to claim 97, wherein the TCR comprises CDR3α containing the amino acid sequence CAAAAPNNNDMRF (SEQ ID NO: 280) and CDR3β containing the amino acid sequence CASSLLAGGSSYEQYF (SEQ ID NO: 285).

99. The TCR according to claim 97 or 98, wherein the TCR comprises the following CDR sequences: CDR1α-DSASNY (SEQ ID NO: 278), CDR2α-IRSNVGE (SEQ ID NO: 279), CDR3α-CAAAAPNNNNDDMRF (SEQ ID NO: 280), CDR1β-MNHEY (SEQ ID NO: 283), CDR2β-SVGAGI (SEQ ID NO: 284), and CDR3β-CASSLLAGGSSYEQYF (SEQ ID NO: 285), or variants thereof having up to three amino acid substitutions, additions, or deletions.

100. The TCR according to any one of claims 97 to 99, wherein the TCR comprises an α-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

101. The TCR according to any one of claims 97 to 100, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 282, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 287 or 288, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

102. The TCR according to any one of claims 97 to 101, wherein the c-MET peptide comprises or consists of the amino acid sequence of YVDPITSI (SEQ ID NO: 277), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

103. The TCR according to claim 1, wherein the immunogenic peptide is a mesothelin (MSLN) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAAYNDYKLSF (SEQ ID NO: 293), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSLQGVNTEAFF (SEQ ID NO: 298), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

104. The TCR according to claim 103, wherein the TCR comprises CDR3α containing the amino acid sequence of CAAYNDYKLSF (SEQ ID NO: 293) and CDR3β containing the amino acid sequence of CASSLQGVNTEAFF (SEQ ID NO: 298).

105. The TCR according to claim 103 or 104, wherein the TCR comprises the following CDR sequences: CDR1α-SSNFYA (SEQ ID NO: 291), CDR2α-MTLNGDE (SEQ ID NO: 292), CDR3α-CAAYNDYKLSF (SEQ ID NO: 293), CDR1β-MNHEY (SEQ ID NO: 296), CDR2β-SMNVEV (SEQ ID NO: 297), and CDR3β-CASSLQGVNTEAFF (SEQ ID NO: 298), or variants thereof having up to three amino acid substitutions, additions, or deletions.

106. The TCR according to any one of claims 103 to 105, wherein the TCR comprises an α-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

107. The TCR according to any one of claims 103 to 106, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 295, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 300 or 301, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

108. The TCR according to any one of claims 103 to 107, wherein the MSLN peptide comprises or consists of the amino acid sequence of KLLGPHVLGV (SEQ ID NO: 290), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

109. The TCR according to claim 1, wherein the immunogenic peptide is a CTD phosphatase subunit 1 (CTDP1) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAMREGTSGTYKYIF (SEQ ID NO: 306), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASSNSASRPEQYV (SEQ ID NO: 311), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

110. The TCR according to claim 109, wherein the TCR comprises CDR3α containing the amino acid sequence of CAMREGTSGTYKYIF (SEQ ID NO: 306) and CDR3β containing the amino acid sequence of CASSNSASRPEQYV (SEQ ID NO: 311).

111. The TCR according to claim 109 or 110, wherein the TCR comprises the following CDR sequences: CDR1α-TSDPSYG (SEQ ID NO: 304), CDR2α-QGSYDQQN (SEQ ID NO: 305), CDR3α-CAMREGTSGTYKYIF (SEQ ID NO: 306), CDR1β-SEHNR (SEQ ID NO: 309), CDR2β-FQNEAQ (SEQ ID NO: 310), and CDR3β-CASSNSASRRPEQYV (SEQ ID NO: 311), or variants thereof having up to three amino acid substitutions, additions, or deletions.

112. The TCR according to any one of claims 109 to 111, wherein the TCR comprises an α-chain variable domain containing the amino acid sequence of SEQ ID NO: 307, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain containing the amino acid sequence of SEQ ID NO: 312, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

113. The TCR according to any one of claims 109 to 112, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 308, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 313 or 314, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

114. The TCR according to any one of claims 109 to 113, wherein the CTDP1 peptide comprises or consists of the amino acid sequence of YLNKEIEEA (SEQ ID NO: 303), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

115. The TCR according to claim 1, wherein the immunogenic peptide is a transformed acid-coiled-coil-containing protein 2 (TACC2) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVSGLGGSNYKLTF (SEQ ID NO: 319), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CASRSGTGTMDEQFF (SEQ ID NO: 324), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

116. The TCR according to claim 115, wherein the TCR comprises CDR3α containing the amino acid sequence CAVSGLGGGSNYKLTF (SEQ ID NO: 319) and CDR3β containing the amino acid sequence CASRSGTGTMDEQFF (SEQ ID NO: 324).

117. The TCR according to claim 115 or 116, wherein the TCR comprises the following CDR sequences: CDR1α-SSVPPY (SEQ ID NO: 317), CDR2α-YTSAATLV (SEQ ID NO: 318), CDR3α-CAVSGLGGGSNYKLTF (SEQ ID NO: 319), CDR1β-MNHNY (SEQ ID NO: 322), CDR2β-SVGAGI (SEQ ID NO: 323), and CDR3β-CASRSGTGTMDEQFF (SEQ ID NO: 324), or variants thereof having up to three amino acid substitutions, additions, or deletions.

118. The TCR according to any one of claims 115 to 117, wherein the TCR comprises an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 320, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 325, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

119. The TCR according to any one of claims 115 to 118, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 321, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 326 or 327, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

120. The TCR according to any one of claims 115 to 119, wherein the TACC2 peptide comprises or consists of the amino acid sequence of YRNSYEIEY (SEQ ID NO: 316), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

121. The TCR according to claim 1, wherein the immunogenic peptide is a prostaglandin F2 receptor negative regulator (PTGFRN) peptide, and the TCR comprises CDR3α, which includes the amino acid sequence of CAVNRDDKIIF (SEQ ID NO: 332), or a variant thereof having up to three amino acid substitutions, additions, or deletions, and CDR3β, which includes the amino acid sequence of CSARDYKQVLRGSYNSPLHF (SEQ ID NO: 337), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

122. The TCR according to claim 121, wherein the TCR comprises CDR3α containing the amino acid sequence of CAVNRDDKIIF (SEQ ID NO: 332) and CDR3β containing the amino acid sequence of CSARDYKQVLRGSYNSPLHF (SEQ ID NO: 337).

123. The TCR according to claim 121 or 122, wherein the TCR comprises the following CDR sequences: CDR1α-DSAIYN (SEQ ID NO: 330), CDR2α-IQSSQRE (SEQ ID NO: 331), CDR3α-CAVNRDDKIIF (SEQ ID NO: 332), CDR1β-DFQATT (SEQ ID NO: 335), CDR2β-SNEGSKA (SEQ ID NO: 336), and CDR3β-CSARDYKQVLRGSYNSPLHF (SEQ ID NO: 337), or variants thereof having up to three amino acid substitutions, additions, or deletions.

124. The TCR according to any one of claims 121 to 123, wherein the TCR comprises an α-chain variable domain comprising the amino acid sequence of SEQ ID NO: 333, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β-chain variable domain comprising the amino acid sequence of SEQ ID NO: 338, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

125. The TCR according to any one of claims 121 to 124, wherein the TCR comprises an α chain containing the amino acid sequence of SEQ ID NO: 334, or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a β chain containing the amino acid sequence of SEQ ID NO: 339 or 340, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

126. The TCR according to any one of claims 121 to 125, wherein the PTGFRN peptide comprises or consists of the amino acid sequence of RLASRPLLL (SEQ ID NO: 329), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

127. A TCR according to any one of claims 1 to 126, comprising one or more mutations at the α-chain / β-chain interface, wherein the frequency of mispairing between the chain and the α-chain and β-chain of the endogenous TCR is reduced when the α-chain and β-chain are expressed in T cells.

128. The TCR according to claim 127, wherein one or more mutations introduce cysteine ​​residues into the constant region domains of the α chain and the β chain, and the cysteine ​​residues can form disulfide bonds between the α chain and the β chain.

129. The TCR according to any one of claims 1 to 128, wherein the TCR includes a mouse-like constant region.

130. The TCR according to any one of claims 1 to 129, wherein the TCR is a soluble TCR.

131. A polynucleotide encoding the α-chain and / or β-chain of a TCR according to any one of claims 1 to 130.

132. The polynucleotide according to claim 131, wherein the polynucleotide encodes the α chain linked to the β chain.

133. The polynucleotide according to claim 131 or 132, wherein the polynucleotide further encodes one or more short interfering RNAs (siRNAs) or other agents that can reduce or prevent the expression of one or more endogenous TCR genes.

134. A vector comprising a polynucleotide according to any one of claims 131 to 133.

135. The vector according to claim 134, wherein the vector is a plasmid or a viral vector.

136. The vector according to claim 134 or 135, wherein the vector comprises one or more polynucleotides encoding CD3 chains, CD8, suicide genes and / or selection markers.

137. A cell comprising a TCR according to any one of claims 1 to 130, a polynucleotide according to any one of claims 131 to 133, or a vector according to any one of claims 134 to 136.

138. The cell according to claim 137, wherein the cell further comprises a vector encoding one or more CD3 chains, CD8, suicide genes and / or selection markers.

139. The cell according to claim 137 or 138, wherein the cell is a T cell, a lymphocyte, or a stem cell.

140. The cell according to claim 139, wherein the T cell, lymphocyte, or stem cell is selected from the group consisting of CD4+ cells, CD8+ cells, naive T cells, memory stem T cells, central memory T cells, double-negative T cells, effector memory T cells, effector T cells, Th0 cells, Tc0 cells, Th1 cells, Tc1 cells, Th2 cells, Tc2 cells, Th17 cells, Th22 cells, gamma / delta T cells, natural killer (NK) cells, natural killer T (NKT) cells, cytokine-induced killer (CIK) cells, hematopoietic stem cells, and pluripotent stem cells.

141. The cell according to any one of claims 137 to 140, wherein the cell is a T cell, and optionally, the cell is a T cell isolated from a subject.

142. A cell according to any one of claims 137 to 141, wherein an endogenous gene encoding a TCRα chain and / or an endogenous gene encoding a TCRβ chain is disrupted, preferably the endogenous gene encoding a TCRα chain and / or the endogenous gene encoding a TCRβ chain is not expressed, and optionally the endogenous gene encoding a TCRα chain and / or the endogenous gene encoding a TCRβ chain is disrupted by insertion of an expression cassette containing a polynucleotide sequence encoding a TCR as described in any one of claims 1 to 130.

143. A cell according to any one of claims 137 to 142, wherein one or more endogenous genes encoding MHC are disrupted.

144. Cells according to any one of claims 137 to 143, wherein endogenous genes involved in persistence, proliferation, activity, resistance to exhaustion / senescence / inhibitory signals, homing ability, or other T cell functions are disrupted, and the endogenous genes involved in persistence, proliferation, activity, resistance to exhaustion / senescence / inhibitory signals, homing ability, or other T cell functions are optionally selected from the group consisting of PD1, TIM3, LAG3, 2B4, KLRG1, TGFbR, CD160, TIGIT, CTLA4, and CD39.

145. A method for preparing cells, comprising the step of introducing a polynucleotide according to any one of claims 131 to 133 or a vector according to any one of claims 134 to 136 into cells in vitro, ex vivo, or in vivo, for example by transfection or transduction.

146. The method according to claim 145, wherein the method comprises a step of T cell editing, which includes disrupting an endogenous gene encoding a TCRα chain and / or an endogenous gene encoding a TCRβ chain with an artificial nuclease, and preferably the artificial nuclease is selected from the group consisting of zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), and the CRISPR / Cas system.

147. The method according to claim 146, wherein the method comprises the step of targeting and incorporating an expression cassette into the endogenous gene encoding the TCRα chain and / or the endogenous gene encoding the TCRβ chain, which has been disrupted by the artificial nuclease, the expression cassette comprising a polynucleotide sequence encoding the TCR according to any one of claims 1 to 130.

148. The method according to any one of claims 145 to 147, wherein the method comprises the step of disrupting one or more endogenous genes encoding MHC, and optionally the cells prepared by the method are non-alloreactive universal T cells.

149. The method according to any one of claims 145 to 148, wherein the method comprises the step of disrupting one or more endogenous genes to modify persistence, proliferation, activity, resistance to exhaustion / aging / inhibitory signals, homing ability, or other T cell function, and optionally the method comprises the step of targeting and incorporating an expression cassette to an endogenous gene involved in persistence, proliferation, activity, resistance to exhaustion / aging / inhibitory signals, homing ability, or other T cell function disrupted by an artificial nuclease, wherein the expression cassette comprises a polynucleotide sequence encoding a TCR according to any one of claims 1 to 130, and optionally the endogenous gene is selected from the group consisting of PD1, TIM3, LAG3, 2B4, KLRG1, TGFbR, CD160, TIGIT, CTLA4, and CD39.

150. A chimeric molecule comprising a TCR or a portion thereof according to any one of claims 1 to 130, conjugated to a noncellular substrate, a toxin and / or an antibody, wherein the noncellular substrate is optionally selected from the group consisting of nanoparticles, exosomes and other noncellular substrates.

151. A pharmaceutical composition comprising a TCR according to any one of claims 1 to 130, a polynucleotide according to any one of claims 131 to 133, a vector according to any one of claims 134 to 136, a cell according to any one of claims 137 to 144, a cell prepared by the method according to any one of claims 145 to 149, or a chimeric molecule according to claim 150.

152. Cells prepared by any one of claims 137 to 144 or by the method described in any one of claims 145 to 149 for use in adoptive cell transfer, preferably adoptive T cell transfer, wherein the adoptive T cell transfer is optionally allogeneic adoptive T cell transfer, autologous adoptive T cell transfer, or universal non-alloreactive adoptive T cell transfer.

153. A TCR according to any one of claims 1 to 130, a polynucleotide according to any one of claims 131 to 133, a vector according to any one of claims 134 to 136, a cell according to any one of claims 137 to 144, a cell prepared by the method according to any one of claims 145 to 149, a chimeric molecule according to claim 150, or a pharmaceutical composition according to claim 151, for use in therapeutic purposes.

154. A TCR according to any one of claims 1 to 130, a polynucleotide according to any one of claims 131 to 133, a vector according to any one of claims 134 to 136, a cell according to any one of claims 137 to 144, a cell prepared by the method according to any one of claims 145 to 149, a chimeric molecule according to claim 150, or a pharmaceutical composition according to claim 151, for use in the treatment and / or prevention of proliferative disorders.

155. The proliferative disorder is a hematological malignancy or a solid tumor, and the hematological malignancy is optionally selected from the group consisting of acute myeloid leukemia (AML), chronic myeloid leukemia (CML), lymphoblastic leukemia, acute lymphoblastic leukemia (ALL), myelodysplastic syndrome, lymphoma, multiple myeloma, non-Hodgkin lymphoma, and Hodgkin lymphoma, and the solid tumor is optionally selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, colon cancer, bile duct cancer, pancreatic cancer A TCR, polynucleotide, vector, cell, chimeric molecule, or pharmaceutical composition for use according to claim 154, selected from the group consisting of visceral cancer, ovarian cancer, head and neck cancer, synovial sarcoma, angiosarcoma, osteosarcoma, thyroid cancer, oral cancer, hepatocellular carcinoma, bladder cancer, endometrial cancer, neuroblastoma, rhabdomyosarcoma, liver cancer, melanoma, prostate cancer, kidney cancer, soft tissue sarcoma, urothelial carcinoma, cholangiocarcinoma, glioblastoma, mesothelioma, cervical cancer, and colorectal cancer.

156. A TCR, polynucleotide, vector, cell, chimeric molecule, or pharmaceutical composition for use according to claim 154, wherein the proliferative disorder is acute myeloid leukemia (AML), or the proliferative disorder is colorectal cancer and / or pancreatic cancer.

157. A method for treating and / or preventing a proliferative disorder, comprising the step of administering a therapeutically effective amount of a TCR according to any one of claims 1 to 130, a polynucleotide according to any one of claims 131 to 133, a vector according to any one of claims 134 to 136, a cell according to any one of claims 137 to 144, a cell prepared by the method according to any one of claims 145 to 149, a chimeric molecule according to claim 150, or a pharmaceutical composition according to claim 151 to a subject in need thereof.

158. The method according to claim 157, wherein the proliferative disorder is a hematological malignancy or a solid tumor, and the hematological malignancy is optionally selected from the group consisting of acute myeloid leukemia (AML), chronic myeloid leukemia (CML), lymphoblastic leukemia, acute lymphoblastic leukemia (ALL), myelodysplastic syndrome, lymphoma, multiple myeloma, non-Hodgkin lymphoma, and Hodgkin lymphoma, and the solid tumor is optionally selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, colon cancer, bile duct cancer, pancreatic cancer, ovarian cancer, head and neck cancer, synovial sarcoma, angiosarcoma, osteosarcoma, thyroid cancer, oral cancer, hepatocellular carcinoma, bladder cancer, endometrial cancer, neuroblastoma, rhabdomyosarcoma, liver cancer, melanoma, prostate cancer, kidney cancer, soft tissue sarcoma, urothelial carcinoma, bile duct cancer, glioblastoma, mesothelioma, cervical cancer, and colorectal cancer.

159. The method according to claim 157, wherein the proliferative disorder is acute myeloid leukemia (AML), or the proliferative disorder is colorectal cancer and / or pancreatic cancer.

160. Use of a TCR according to any one of claims 1 to 130, a polynucleotide according to any one of claims 131 to 133, a vector according to any one of claims 134 to 136, a cell according to any one of claims 137 to 144, a cell prepared by the method according to any one of claims 145 to 149, a chimeric molecule according to claim 150, or a pharmaceutical composition according to claim 151 in the manufacture of a pharmacopoeia for the treatment and / or prevention of proliferative disorders.

161. The use according to claim 160, wherein the proliferative disorder is a hematological malignancy or a solid tumor, and the hematological malignancy is optionally selected from the group consisting of acute myeloid leukemia (AML), chronic myeloid leukemia (CML), lymphoblastic leukemia, acute lymphoblastic leukemia (ALL), myelodysplastic syndrome, lymphoma, multiple myeloma, non-Hodgkin lymphoma, and Hodgkin lymphoma, and the solid tumor is optionally selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, colon cancer, bile duct cancer, pancreatic cancer, ovarian cancer, head and neck cancer, synovial sarcoma, angiosarcoma, osteosarcoma, thyroid cancer, oral cancer, hepatocellular carcinoma, bladder cancer, endometrial cancer, neuroblastoma, rhabdomyosarcoma, liver cancer, melanoma, prostate cancer, kidney cancer, soft tissue sarcoma, urothelial carcinoma, bile duct cancer, glioblastoma, mesothelioma, cervical cancer, and colorectal cancer.

162. The use according to claim 160, wherein the proliferative disorder is acute myeloid leukemia (AML), or the proliferative disorder is colorectal cancer and / or pancreatic cancer.

163. Isolated immunogenic cathepsin G (CTSG) peptides comprising or consisting of amino acid sequences selected from the following: GIVSYGKSSGVPEV (SEQ ID NO: 58), SXXXXPXVFTRVSSFL (SEQ ID NO: 535), SGVPPEVFTRVSSFL (SEQ ID NO: 81), TMRSFKLLLDQMETPL (SEQ ID NO: 95), and their variants each having up to three amino acid substitutions, additions, or deletions, or immunogenic fragments thereof.

164. The isolated immunogenic CTSG peptide according to claim 163, wherein the CTSG peptide comprises or consists of GIVSYGKSSGVPEPEV (SEQ ID NO: 58), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof.

165. The isolated immunogenic CTSG peptide according to claim 163, wherein the CTSG peptide comprises or consists of XXXPXVFTRVSSFL (SEQ ID NO: 535), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, preferably the CTSG peptide comprises or consists of the amino acid sequence of SGVPPEVFTRVSSFL (SEQ ID NO: 81), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, preferably the CTSG peptide comprises or consists of the amino acid sequence of VFTRVSSFL (SEQ ID NO: 82), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or the CTSG peptide comprises or consists of the amino acid sequence of VPPEVFTRVSSFL (SEQ ID NO: 83), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

166. The isolated immunogenic CTSG peptide according to claim 163, wherein the CTSG peptide comprises or consists of TMRSFKLLDQMETPL (SEQ ID NO: 95), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof.

167. The isolated immunogenic CTSG peptide according to any one of claims 163 to 166, wherein the immunogenic fragment has a length of 9 amino acids or more, and preferably the immunogenic fragment is a 9-mer fragment.

168. Isolated immunogenic cyclin A1 (CCNA1) peptides comprising or consisting of the following: TEYAEEIYQYL (SEQ ID NO: 114), PETLAAFTGYSLSEI (SEQ ID NO: 128), AELSLLEADPFLKYL (SEQ ID NO: 153), LLEADPFLKYLPSLI (SEQ ID NO: 527), WEGPGLPDFVF (SEQ ID NO: 172), KRQLLKMEHLLLLKVL (SEQ ID NO: 185), PETLAAFTGYSLSEI (SEQ ID NO: 201), and amino acid sequences selected from their variants each having up to three amino acid substitutions, additions, or deletions, or immunogenic fragments thereof.

169. The isolated immunogenic CCNA1 peptide according to claim 168, wherein the CCNA1 peptide comprises or consists of TEYAEEIYQYL (SEQ ID NO: 114), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, preferably the CCNA1 peptide comprises or consists of the amino acid sequence of YAEEIYQYL (SEQ ID NO: 115), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or the CCNA1 peptide comprises or consists of the amino acid sequence of EYAEEIYQY (SEQ ID NO: 116), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

170. The isolated immunogenic CCNA1 peptide according to claim 168, wherein the CCNA1 peptide comprises or consists of PETLAAFTGYSLSEI (SEQ ID NO: 128), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, preferably the CCNA1 peptide comprises or consists of the amino acid sequence of ETLAAFTGY (SEQ ID NO: 129), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or the CCNA1 peptide comprises or consists of the amino acid sequence of LAAFTGYSL (SEQ ID NO: 130), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

171. The isolated immunogenic CCNA1 peptide according to claim 168, wherein the CCNA1 peptide comprises or consists of AELSLLEADPFLKYL (SEQ ID NO: 153), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, or the CCNA1 peptide comprises or consists of LLEADPFLKYLPSLI (SEQ ID NO: 527), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, preferably the CCNA1 peptide comprises or consists of the amino acid sequence of EADPFLKYL (SEQ ID NO: 154), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

172. The isolated immunogenic CCNA1 peptide according to claim 168, wherein the CCNA1 peptide comprises or consists of WEGPGLPFVF (SEQ ID NO: 172), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, preferably the CCNA1 peptide comprises or consists of the amino acid sequence of GPGLPFVF (SEQ ID NO: 173), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

173. The CCNA1 peptide comprises or consists of KRQLLKMEHLLLKVL (SEQ ID NO: 185), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof. Preferably, the CCNA1 peptide comprises or consists of the amino acid sequence of RQLLKMEHL (SEQ ID NO: 186), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or the CCNA1 peptide comprises the amino acid sequence of MEHLLLKVL (SEQ ID NO: 187), or up to three amino acids The isolated immunogenic CCNA1 peptide according to claim 168, comprising or consisting of a variant having substitutions, additions, or deletions thereof, or the CCNA1 peptide comprising or consisting of the amino acid sequence of MEHLLLKV (SEQ ID NO: 188) or a variant having up to three amino acid substitutions, additions, or deletions thereof, or the CCNA1 peptide comprising or consisting of the amino acid sequence of KMEHLLLKV (SEQ ID NO: 189) or a variant having up to three amino acid substitutions, additions, or deletions thereof.

174. The isolated immunogenic CCNA1 peptide according to claim 168, wherein the CCNA1 peptide comprises or consists of PETLAAFTGYSLSEI (SEQ ID NO: 201), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or an immunogenic fragment thereof, preferably the CCNA1 peptide comprises or consists of the amino acid sequence of ETLAAFTGY (SEQ ID NO: 202), or a variant thereof having up to three amino acid substitutions, additions, or deletions, or the CCNA1 peptide comprises or consists of the amino acid sequence of LAAFTGYSL (SEQ ID NO: 203), or a variant thereof having up to three amino acid substitutions, additions, or deletions.

175. The isolated immunogenic CCNA1 peptide according to any one of claims 168 to 174, wherein the immunogenic fragment has a length of 9 amino acids or more, and preferably the immunogenic fragment is a 9-mer fragment.

176. A T cell receptor (TCR) that binds to an immunogenic CTSG peptide when presented by a major histocompatibility complex (MHC), wherein the immunogenic CTSG peptide is as described in claim 165.

Images