Specific T cell receptors for mutant MYD88 in adoptive T cell therapy L265P Protein epitopes

Abstract

The present invention relates to the technical field of immunotherapy, and in particular to adoptive T cell therapy or T cell receptor (TCR) gene therapy. The present invention provides a nucleic acid encoding at least one TCRα or β chain construct of a TCR construct, the TCR construct being capable of specifically binding to the MYD88L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02 having a high affinity for the peptide / HLA complex. The present invention also provides the corresponding protein and host cell of the above nucleic acid, preferably CD8+ T cells, as well as the medical uses of the nucleic acid, protein or host cell, particularly in the diagnosis, prevention and / or treatment of cancers expressing MYD88L265P, such as non-Hodgkin B cell lymphomas selected from diffuse large B cell lymphoma (DLBCL) (such as activated B cell type DLBCL (ABC-DLBCL) or primary CNS DLBCL, cutaneous DLBCL, leg type DLBCL or testicular DLBCL), lymphoplasmacytic lymphoma (LPL) (such as Waldenström macroglobulinemia (WM)) and IgM monoclonal gammopathy of undetermined significance (IgM MGUS).

Description

Technical Field

[0001] The present invention relates to the technical field of immunotherapy, and in particular to adoptive T cell therapy or T cell receptor (TCR) gene therapy. The present invention provides a nucleic acid encoding at least one TCRα or β chain construct of a TCR construct, which TCR construct is capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02 having a high affinity for the peptide / HLA complex. The present invention also provides the corresponding protein and host cell of the above nucleic acid, preferably CD8+ T cells, and the medical uses of the nucleic acid, protein or host cell, especially in the diagnosis, prevention and / or treatment of cancers carrying the MYD88 L265P mutation. The cancers carrying the MYD88 L265P mutation are, for example, selected from non-Hodgkin B cell lymphomas such as diffuse large B cell lymphoma (DLBCL) (such as activated B cell type DLBCL (ABC-DLBCL) or primary CNS DLBCL, cutaneous DLBCL, leg type DLBCL or testicular DLBCL), lymphoplasmacytic lymphoma (LPL) (such as Waldenström macroglobulinemia (WM)) and IgM monoclonal gammopathy (IgM MGUS). Background Art

[0002] B cell-derived tumors remain one of the leading causes of death in the Western world. It is estimated that there are approximately 1,500 to 2,000 new cases of high-grade B cell lymphoma in Germany each year. Up to 40% of these patients will relapse after initial standard treatment or have no response at all, indicating an urgent need for alternative treatment options. The incidence of lymphoma increases sharply with age, and the prognosis is even worse for many patients aged 75 or older.

[0003] Although chemotherapy has limitations in terms of toxicity and the development of drug resistance, it remains the main treatment option for most cancer types. A combination of multiple chemotherapy regimens and the monoclonal antibody rituximab targeting the CD20 B cell antigen is widely used as the first-line treatment for diffuse large B cell lymphoma, with a cure rate of approximately 60%. Even with high-dose chemotherapy with stem cell rescue, less than one-third of patients with relapsed / refractory disease can be rescued after first-line treatment. The prognosis of primary CNS lymphoma is even worse: only high-dose chemotherapy seems to be effective, but due to age limitations and comorbidities, it is only feasible in a small number of patients because it occurs more frequently in patients over 70 years old.

[0004] A chimeric antigen receptor (CAR) is a chimera of the antigen-binding domain of an antibody that can recognize a cell surface antigen that binds to the TCR domain. T cells engineered to express a CAR thus target cells expressing the antigen to which the CAR binds, regardless of any HLA restriction.

[0005] CART cells targeting CD19 have been successfully demonstrated to be applicable in the treatment of approximately 50% of patients with refractory and relapsed DLBCL, also demonstrating the efficacy of adoptive T cell therapy. In recent years, clinical studies of adoptive T cell therapy (ATT) using chimeric antigen receptor gene transfer against the B cell antigen CD19 have achieved remarkable success and are known as "breakthrough cancer therapies". Most researchers are developing studies on this same strategy, mainly by targeting B cell lineage antigens such as CD19, CD20, and CD22. However, tumor escape through modulation of the surface expression of the target antigen is a major limitation of this strategy, resulting in recurrence in at least 50% of treated patients and its high cost. In addition, CAR-based adoptive T cell therapy can only target cell surface proteins and not intracellular proteins. Although more specific than chemotherapy, CAR-based ATT is not truly tumor-specific because after B cell-directed CAR-ATT, the entire B cell compartment, including malignant and normal B lymphocytes, is cleared, which often leads to severe B cell depletion and may require long-term immunoglobulin replacement.

[0006] The TCR is a heterodimeric cell surface protein of the immunoglobulin superfamily that is associated with invariant proteins of the CD3 complex involved in mediating signal transduction. The TCR exists in the forms of αβ and γδ, which are similar in structure but have very different structural locations and possible functions. The α and β chains of the native heterodimeric αβ TCR are transmembrane proteins, each containing two extracellular domains, namely the membrane-proximal constant domain and the membrane-distal variable domain. Each constant and variable region contains an intrachain disulfide bond. The variable region contains highly polymorphic loops similar to the complementarity-determining regions (CDRs) of antibodies.

[0007] The variable region of each TCR chain includes a variable segment and a joining segment, and in the case of the β chain, also a diversity segment. Each variable region consists of three CDRs (complementary determining regions) embedded in a framework sequence, one of which is a hypervariable region called CDR3. Multiple types of α-chain variable regions (Vα) and multiple types of β-chain variable regions (Vβ) are distinguished by their framework, CDR1 and CDR2 sequences, and partially defined CDR3 sequences. Specific TRAV or TRBV numbers are assigned to Vα or Vβs by the IMGT nomenclature. The specificity of the T cell receptor for the recognized epitope is mainly determined by the CDR3 region (Danska et al., 1990. J. Exp. Med. 172:27 - 33; Garcia et al., 2005. Cell 122(3):333 - 336).

[0008] Adoptive TCR gene therapy can endow a patient's own T cells with the required specificity and generate a sufficient number of T cells in a short time, thus avoiding their exhaustion. TCRs can be transduced into all T cells or T cell subsets, such as CD8, central memory T cells, or T cells with stem cell characteristics, which can ensure better persistence and function upon transfer. TCR-engineered T cells can be infused into cancer patients who have lymphopenia caused, for example, by chemotherapy or radiotherapy, inducing homeostatic expansion, thereby greatly enhancing the engraftment and long-term persistence of the transplanted T cells with a high cure rate.

[0009] In contrast to CAR-based strategies, TCR-based adoptive T cell therapy relies on the recognition of TCRs of processed epitopes of antigens that conventionally exist in the context of MHC molecules, rather than antibody recognition as in CARs. The advantage is that surface expression is not required for TCR recognition, and thus, modulating surface antigen expression upon CAR binding does not pose a limitation. In addition, cancer mutations mainly occur in cytoplasmic proteins that regulate cell proliferation and survival or are sensitive to drugs and other regulatory signals, rather than in surface molecules: T cell receptors can target any protein independent of cell localization, thus greatly broadening the range of targetable antigens, which include lineage-specific surface antigens (such as CARs) and true tumor-specific intracellular antigens.

[0010] Ideally, cancer-specific mutant antigens derived from somatic mutations acquired during tumor development, so-called "neoantigens", are the best possible targets for immune system recognition because they are strictly expressed by cancer cells, which implies higher specificity and lower off-target toxicity. Cancers carrying oncogenic driver mutations are still very attractive for TCR gene therapy if the underlying mutations result in abnormal peptides on high-affinity MHC molecules (Blankenstein et al., 2015. Curr Opin Immunol. 33:112-119).

[0011] MYD88 is an intracellular adaptor protein. A missense mutation that changes leucine 265 to proline (L265P) in MYD88 is one of the most common driver mutations and can be found in approximately one-fifth of all lymphoid malignancies, and is more common in aggressive and treatment-resistant cases. The mutation occurs at a high frequency in B-cell lymphomas, such as in diffuse large B-cell lymphoma, such as activated B-cell type DLBCL (ABC-DLBCL) or primary CNS DLBCL, cutaneous DLBCL, leg-type DLBCL or testicular DLBCL; lymphoplasmacytic lymphoma (LPL), such as IgM monoclonal gammopathy and approximately 90% of patients with Waldenström macroglobulinemia (WM) (Yu et al., 2018. Cancer Res. 78(10):2457-62; Knittel et al., 2016. Blood 127(22):2732-2741; Rovira et al., 2016. Clin Cancer Res1-10; Lee et al., 2017. Scientific Reports 7:1785).

[0012] Peptides containing the MYD88 L265P mutation have been proposed for use in cancer immunotherapy (patent DE102015106731A1, Nelde et al., 2017. Oncoimmunology 6(3):e1219825). Based on in silico predictions, Nelde et al. (2017) identified potential MYD88 L265P containing HLA ligands for multiple HLA class I restrictions. A set of HLA-I MYD88L265P-derived ligands were shown to elicit specific cytotoxic T cell responses against HLA-B*07 and HLA-B*15, and Nelde et al. investigated whether the peptides could occur naturally.

[0013] Nielsen et al. (2017. Oncommunology 6(7):e1321184) evaluated the recognition of common driver mutations, such as the MYD88 L265P mutation, by T cells from healthy donors by screening libraries of all possible 8-, 9-, 10-, and 11-mer mutant peptides on donor T cells. They identified CD8+ T cells specific for the peptide RPIPIKYKA (SEQ ID NO:1, where the bold P denotes the L265P mutation) of MYD88 L265P presented by HLA-B*07:02 in one donor, and found evidence that the peptide could also be processed in human B cells. Other peptides, particularly the longer peptide RPIPIKYKAM (SEQ ID NO:2), were also recognized by donor T cells on target cells pulsed with the peptide, but most T cell lines reactive to the peptide failed to recognize B cells transfected with MYD88 L265P. The authors thus concluded that 75% of the candidate peptides were not naturally processed, which would render TCRs recognizing the peptide unsuitable for T cell therapy. Regarding the remaining T cell lines reactive to the SEQ ID NO:1 peptide that could be processed, Nielsen et al. discussed options for TCR gene therapy, but they considered the utility of TCR engineering to be limited based on the small number of patients expressing HLA-B*07. The T cells were not cloned, and TCR sequences were not provided. The authors thus recommended moving to an expanded list of alternative target antigens that frequently carry putative driver mutations in lymphoma. Additionally, the authors taught the use of peptides for therapeutic purposes by vaccination. SUMMARY OF THE INVENTION

[0014] In view of the above, the inventors solved the problem of providing advantageous TCR constructs that are capable of specifically targeting peptides containing amino acid substitutions due to driver gene mutations in B cell lymphoma, which can be naturally processed and presented on HLA, and which, preferably, have a high affinity that allows the TCR constructs to be used for therapeutic purposes. This problem is solved by the subject matter of the claims.

[0015] The inventors provide TCR constructs that recognize the epitope of the antigen in MYD88 L265P, i.e., TCR constructs that bind the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02, and which TCRs have surprisingly high affinity. Contrary to the teachings of Nielsen et al., they also found that MYD88 L265P can be naturally processed to generate the peptide of SEQ ID NO:2 in the presence of HLA-B*07:02, and that T cells targeting the peptide can thus be advantageously used for the treatment of tumors expressing MYD88 L265P.

[0016] The present invention provides a nucleic acid, wherein the nucleic acid encodes at least one TCRα chain or β chain construct of a TCR construct that can specifically bind to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02,

[0017] a) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:13, and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:16; or

[0018] b) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:23,

[0019] and / or wherein the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:24, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:25, and the CDR3 sequence of SEQ ID NO:26; or

[0020] c) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:33,

[0021] and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:36; or

[0022] d) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:43,

[0023] and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:46, or

[0024] e) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:93,

[0025] and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:96, or

[0026] f) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:103,

[0027] and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:106; or

[0028] g) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:113,

[0029] and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:116; or

[0030] h) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:123,

[0031] and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:126; or

[0032] i) wherein the TCRα chain construct comprises the CDR3 sequence of SEQ ID NO:133,

[0033] and / or wherein the TCRβ chain construct comprises the CDR3 sequence of SEQ ID NO:136.

[0034] Since the affinity and specificity can be further optimized by methods known in the art described below, the present invention also provides a nucleic acid that can specifically bind to at least one TCRα chain or β chain construct of a TCR construct that specifically binds to the MYD88L265P peptide of SEQ ID NO:2 in the presence of HLA - B*07:02,

[0035] a) wherein the TCRα chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO:13, and / or wherein the TCRβ chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO:16; or

[0036] b) wherein the TCRα chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO:23, and / or wherein the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:24, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:25, and a CDR3 sequence having at least 90% sequence identity with SEQ ID NO:26; or

[0037] c) wherein the TCRα chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO:33, and / or wherein the TCRβ chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO:36; or

[0038] d) wherein the TCR α-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 43, and / or wherein the TCR β-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 46, or

[0039] e) wherein the TCR α-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 93, and / or wherein the TCR β-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 96, or

[0040] f) wherein the TCR α-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 103, and / or wherein the TCR β-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 106; or

[0041] g) wherein the TCR α-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 113, and / or wherein the TCR β-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 116; or

[0042] h) wherein the TCR α-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 123, and / or wherein the TCR β-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 126; or

[0043] i) wherein the TCR α-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 133, and / or wherein the TCR β-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 136.

[0044] The TCR α- and / or chain constructs in e) of the preferred TCR constructs may be parts comprising the variable region of TCR2304.

[0045] The TCR α- and / or chain constructs in a) of the preferred TCR constructs may be parts comprising the variable region of TCR2207.

[0046] The TCRα and / or chain construct in b) of the preferred TCR construct may be a part containing the variable region of TCR2205. It should be noted that in the presence of the TCRα and / or chain construct in b) above, the CDR3 sequence of the β chain of the TCR2205, i.e., SEQ ID NO:26, has been previously published in the article titled "Tissue distribution ad clonaldiversity of the T and B-cell repertoire in type 1diabetes in the supplementary data, Seay et al., 2016, JCI Insight.1(20):e88242". The remaining parts of the gene are especially different in CDR1 and CDR2, and the variant gene subtypes and the corresponding α chain sequences are different.

[0047] The TCRα and / or chain construct in c) of the preferred TCR construct may be a part containing the variable region of TCR1610.

[0048] The TCRα and / or chain construct in d) of the preferred TCR construct may be a part containing the variable region of TCR1605.

[0049] The TCRα and / or chain construct in f) of the preferred TCR construct may be a part containing the variable region of TCR2705.

[0050] The TCRα and / or chain construct in g) of the preferred TCR construct may be a part containing the variable region of TCR2709.

[0051] The TCRα and / or chain construct in h) of the preferred TCR construct may be a part containing the variable region of TCR2716.

[0052] The TCRα and / or chain construct in i) of the preferred TCR construct may be a part containing the variable region of TCR2719.

[0053] In the presence of HLA-B*07:02, all TCR constructs of the present invention are capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2. The inventors can demonstrate that the histocompatibility antigen HLA-B7:02 can effectively present this mutation for T cell receptor recognition, and it is a relatively common MHC haplotype, with an occurrence frequency of 15 - 25% in Germans and approximately 30% in North American Caucasians.

[0054] One of the TCR constructs of the present invention, wherein the TCR α-chain construct comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 13, and wherein the TCR β-chain construct, such as TCR2207, comprises a CDR3 sequence having at least 90% sequence identity with SEQ ID NO: 16, which is also capable of specifically recognizing 9-mer, 11-mer and 12-mer MYD88 L265P peptides, in particular the peptides of SEQ ID NO: 1, SEQ ID NO: 3 or SEQ ID NO: 4.

[0055] The TCR constructs can specifically recognize the peptide of SEQ ID NO: 2. In particular, they do not recognize the corresponding MYD88 wild-type peptide of SEQ ID NO: 3. Preferably, they also do not have significant cross-reactivity with non-MYD88 L265P self-peptides, in particular self-peptides that occur on the HLA of the patient to be treated with TCR.

[0056] As used herein, the terms "capable of specifically binding", "recognizing", or "specific for" a given antigen mean that the TCR construct can specifically bind and immunologically recognize the epitope and HLA, more preferably with high affinity. For example, in the presence of HLA*B07:02, if T cells expressing the TCR secrete at least about 200 pg / ml or more (e.g., 250 pg / ml or higher, 500 pg / ml or higher, 750 pg / ml or higher, 1000 pg / ml or higher, 2000 pg / ml or higher, 2500 pg / ml or higher, 5000 pg / ml or higher) of interferon-γ (IFNγ) when co-cultured with target cells pulsed with low concentrations of the respective peptides (e.g., about 10 -11 M, 10 -10 M, 10 -9 M, 10 - 8 M, 10 -7 M, 10 -6 M, 10 -5 M) (but with the epitope or with an irrelevant control peptide epitope or the wild-type MYD88 peptide of SEQ ID NO: 3), then the TCR will be considered to be capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO: 2. The "specificity" described above can be analyzed by, for example, ELISA.

[0057] In the context of the present invention, "about" should be understood to mean the specified value + / - 10%, preferably + / - 5%.

[0058] Affinity (or avidity, as a typical TCR has two binding sites) can be analyzed by methods well-known to those skilled in the art, e.g., by BiaCore, by staining with MHC-peptide multimers and analyzing the mean fluorescence intensity (MFI) on FACS, or preferably by non-linear curve analysis of IFNγ response, where affinity is inversely proportional to the K D value, as shown in Example 3 or Figure 3 A or C herein. A TCR affinity with a K D value of 10 -7 mol(M) or lower is considered high affinity. Preferably, throughout the invention, the TCR encoded by the TCR construct has a K D value of 7.4×10 -9 M or lower for the peptide of SEQ ID NO:2 in the presence of HLA-B*07:02, where the avidity is more preferably about 2.4×10 -9 M or lower. Such an affinity has been demonstrated to be the affinity between the TCR constructs with two antigen-binding sites of the present invention ( Figure 3 A and 3C).

[0059] In one embodiment, in the nucleic acids of the present invention, the encoded TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:91, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:92, and a CDR3 sequence of SEQ ID NO:93, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:94, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:95, and a CDR3 sequence of SEQ ID NO:96.

[0060] Preferably, in the nucleic acids of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO:97, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO:98. The nucleic acids may comprise SEQ ID NO:99 and SEQ ID NO:100 encoding the variable regions respectively, which represent nucleic acids codon-optimized for expression in human cells.

[0061] In one embodiment, in the nucleic acid of the present invention, the TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 11, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 12, and a CDR3 sequence of SEQ ID NO: 13, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 14, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 15, and a CDR3 sequence of SEQ ID NO: 16.

[0062] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 17, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 18. The nucleic acid may comprise SEQ ID NO: 19 and SEQ ID NO: 20 encoding the variable regions respectively, which represent codon-optimized nucleic acids.

[0063] In one embodiment, in the nucleic acid of the present invention, the TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 21, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 22, and a CDR3 sequence of SEQ ID NO: 23, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 24, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 25, and a CDR3 sequence of SEQ ID NO: 26.

[0064] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 27, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 28. The nucleic acid may comprise SEQ ID NO: 29 and SEQ ID NO: 30 encoding the variable regions respectively, which represent codon-optimized nucleic acids.

[0065] In one embodiment, in the nucleic acid of the present invention, the TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:31, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:32, and a CDR3 sequence of SEQ ID NO:33, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:34, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:35, and a CDR3 sequence of SEQ ID NO:36.

[0066] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO:37, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO:38. The nucleic acid may comprise SEQ ID NO:39 and SEQ ID NO:40 encoding the variable regions respectively, which represent codon-optimized nucleic acids.

[0067] In one embodiment, in the nucleic acid of the present invention, the TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:41, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:42, and a CDR3 sequence of SEQ ID NO:43, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO:44, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO:45, and a CDR3 sequence of SEQ ID NO:46.

[0068] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO:47, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO:48. The nucleic acid may comprise SEQ ID NO:49 and SEQ ID NO:50 encoding the variable regions respectively, which represent codon-optimized nucleic acids.

[0069] In one embodiment, in the nucleic acid of the present invention, the encoded TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 101, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 102, and a CDR3 sequence of SEQ ID NO: 103, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 104, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 105, and a CDR3 sequence of SEQ ID NO: 106.

[0070] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 107, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 108. The nucleic acid may comprise SEQ ID NO: 109 and SEQ ID NO: 110 encoding the variable regions respectively, which represent nucleic acids codon-optimized for expression in human cells.

[0071] In one embodiment, in the nucleic acid of the present invention, the encoded TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 111, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 112, and a CDR3 sequence of SEQ ID NO: 113, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 114, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 115, and a CDR3 sequence of SEQ ID NO: 116.

[0072] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 117, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 118. The nucleic acid may comprise SEQ ID NO: 119 and 120 encoding the variable regions respectively, which represent nucleic acids codon-optimized for expression in human cells.

[0073] In one embodiment, in the nucleic acid of the present invention, the encoded TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 121, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 122, and a CDR3 sequence of SEQ ID NO: 123, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 124, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 125, and a CDR3 sequence of SEQ ID NO: 126.

[0074] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 127, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 128. The nucleic acid may comprise SEQ ID NO: 129 and SEQ ID NO: 130 encoding the variable regions respectively, which represent nucleic acids codon-optimized for expression in human cells.

[0075] In one embodiment, in the nucleic acid of the present invention, the encoded TCRα chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 131, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 132, and a CDR3 sequence of SEQ ID NO: 133, and / or the TCRβ chain construct comprises a CDR1 sequence having at least 90% sequence identity with SEQ ID NO: 134, a CDR2 sequence having at least 90% sequence identity with SEQ ID NO: 135, and a CDR3 sequence of SEQ ID NO: 136.

[0076] Preferably, in the nucleic acid of the present invention, the TCRα chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 137, and / or the TCRβ chain construct comprises a variable region having at least 90%, preferably at least 95% or 100% sequence identity with SEQ ID NO: 138. The nucleic acid may comprise SEQ ID NO: 139 and SEQ ID NO: 140 encoding the variable regions respectively, which represent nucleic acids codon-optimized for expression in human cells.

[0077] The TCRα chain and / or β chain construct may have Figure 2Features of the TCR 2304, TCR2207, TCR2205, TCR1610 or TCR1605, or TCR2705, TCR2709, TCR2716 or TCR2719 shown. In one embodiment, the TCR construct of the present invention does not contain a β-chain containing TRBV28.

[0078] Preferably, the nucleic acids of the present invention encode a TCR α-chain construct and a TCR β-chain construct. In the context of the present invention, "a" should be understood to mean "one or more" unless otherwise expressly stated. Thus, for example, since the TCR construct of the present invention contains both an α-chain and a β-chain construct, it can be encoded by one or two nucleic acids. The α-chain and β-chain constructs together are capable of specifically binding to the MYD88 L265P peptide in the presence of HLA-B*07:02. As intermediate products, the α-chain and β-chain constructs or the nucleic acids encoding them are also the subject of the present invention.

[0079] Preferably, in all TCR and / or β-chain constructs of the present invention, the sequence identity to the CDR regions defined herein is 100%.

[0080] However, based on the defined CDR3 and variable region sequences provided by the present invention, affinity maturation of the TCR sequence can be performed (Chervin et al., 2008. J Immunol Methods. 339(2):175–84); Robbins et al., 2008. J Immunol. 180:6116–31). Substitutions of non-synonymous nucleotides that result in amino acid exchanges in the CDR3 sequence may lead to an enhanced affinity of the TCR for the target antigen. In addition, changes in the TCR sequence in other parts of the variable TRA and TRB regions may alter the affinity of the TCR for the peptide-MHC complex. This may increase the overall affinity of the TCR for peptide-MHC, but there is a risk of increased non-specific recognition and cross-reactivity (Linette et al., 2013. Blood 122(6):863–72). Preferably, TCRs different from the provided specific sequences retain exclusive specificity for the provided target antigen, i.e., they do not cross-react, and most importantly, they do not cross-react with human self-peptides. The potential cross-reactivity of TCRs can be tested against known self-peptides loaded on cells bearing the correct MHC allele (Morgan et al., 2013, J. Immunother. 36, 133-151). Thus, preferably, adoptively transferred T cells expressing the TCR constructs of the present invention have no negative impact on healthy tissues.

[0081] The TCR α-chain and / or β-chain constructs of the present invention may contain all the features or domains corresponding to their natural pairings, but this is not necessary. Preferably, the TCR α-chain and / or β-chain constructs contain at least one variable region or a variable region and a constant region, and the variable and / or constant regions have at least 60%, at least 70%, at least 80%, at least 90% or at least 95% sequence identity with the human variable or constant TCR regions. For adoptive TCR therapy, it is preferred that the TCR construct contains the full-length TCR α-chain and β-chain, which contain variable regions, constant regions and transmembrane regions. Preferably, the TCR construct is substantially or specifically derived from humans to minimize immunogenicity. The human TCR and β constant regions are, for example, shown in SEQ ID NO:7 (α) and SEQ ID NO:10 (β, TCRBC2, or TCRRBC1 can also be used). However, to prevent pairing with endogenous TCR chains, the constructs of the present invention preferably contain one or more, for example, 1-5, 1-10 or 1-20 amino acid exchanges, insertions or deletions compared to the human sequence, for example, providing additional cysteines to enable the formation of additional disulfide bonds (Sommermeyer et al., 2010, J. Immunol. 184, 6223-31). The constant region of such TCR can be minimally "murinized" by replacing several (usually 9) amino acids of the human constant region sequence with murine sequences (for example, SEQ ID NO:6 (α) and SEQ ID NO:9 (β)). The constant regions of the TCR α-chain and β-chain constructs can also be murine constant regions (SEQ ID NO:5 (α) and SEQ ID NO:8 (β, TCRBC2, or TCRRBC1 can also be used)). The constant regions of the α-chain and β-chain are of the same type, for example, both can be minimally murinized.

[0082] The construct can also be a chimeric antigen receptor or a part thereof, wherein, for example, the human TCR variable region can be linked to a different immunoglobulin constant region, for example, an IgG constant domain, or an antibody domain capable of specifically binding an antigen (such as the CD3 T cell antigen).

[0083] The constructs of the present invention include single-chain constructs (scTCR) and heterodimeric TCR constructs. The scTCR can contain the variable region of the first TCR chain construct (for example, the α-chain) and the entire (full-length) second TCR chain (for example, the β-chain), and vice versa. In addition, the scTCR can optionally contain one or more linkers that link two or more polypeptides together. The linker can be, for example, a peptide that links two single chains as described herein. The present invention also provides such scTCR of the present invention fused with a cytokine, for example, a human cytokine such as IL-2, IL-7 or IL-15.

[0084] The TCR constructs of the present invention can also be provided in the form of multimeric complexes, which comprise at least two scTCR molecules, wherein each of the scTCR molecules is fused to at least one biotin moiety, and wherein the scTCRs are interconnected via biotin-streptamer interactions to form the multimeric complex. The present invention also provides more advanced multimeric complexes, which comprise more than two (e.g., four) scTCRs.

[0085] The TCR constructs of the present invention can be modified to comprise a detectable label, such as a radioisotope, a fluorophore (e.g., fluorescein isothiocyanate (FITC), phycoerythrin (PE)), an enzyme (e.g., alkaline phosphatase, horseradish peroxidase), and particles (e.g., gold particles or magnetic particles).

[0086] The nucleic acids of the present invention, particularly if the nucleic acids encode at least one TCRα chain and one TCRβ chain construct of a TCR construct, can be, for example, vectors that allow the expression of the encoded protein in a host cell, such as a human T cell, such as a viral vector, a transposon, or a vector suitable for CRISPR / CAS-based recombination ((Legut et al., 2018. Blood 131:311-322; Eyquem et al., 2017. Nature 543:113-117; Roth et al., 2018. Nature 559:405-409). In one embodiment, the vector allows integration of the nucleic acid into the host genome.

[0087] Preferably, the TCR α-chain construct and / or TCR β-chain construct or TCR construct of the present invention is a vector. Suitable vectors include vectors designed for propagation and amplification or for expression or both, such as plasmids and viruses. The vector can be an expression vector suitable for expression, and the expression vector is a host cell selected from human T cells or human T cell precursors, preferably human T cells such as CD8+ T cells, CD8+ central memory T cells, CD8+ effector memory T cells, CD8+ stem cell-like T cells. The vector can be a viral vector, for example, a retrovirus, especially a γ-retrovirus or a lentiviral vector. Examples of suitable expression vectors include the retroviral vector MP71. The recombinant expression vector contains regulatory sequences, such as transcription and translation start and stop codons, regulatory untranslated regions, internal ribosome entry sites, and the regulatory sequences are specific to the type of host cell (such as bacteria, fungi, plant or animal cells, for example, human CD8+ T cells as defined above), into which the vector will be introduced and the expression of the nucleic acid of the present invention will occur in this cell. In addition, the vector of the present invention can include one or more marker genes that allow selection of the transformed or transfected host. The recombinant expression vector can contain a natural or preferably heterologous promoter, which is operably linked to the nucleotide sequence encoding the construct of the present invention, or linked to a nucleotide sequence complementary or hybridizing to the nucleotide sequence encoding the construct of the present invention. The selection of the promoter includes, for example, strong promoters, weak promoters, inducible promoters, tissue-specific promoters, and development-specific promoters. The promoter can be a non-viral promoter or a viral promoter. Preferably, it is a heterologous promoter, that is, a promoter that is not naturally linked to the TCR in human T cells, such as the long terminal repeat promoter, which is suitable for expression in human T cells. The recombinant expression vector of the present invention can be designed for transient expression, stable expression, or both. In addition, the recombinant expression vector can be prepared for constitutive expression or inducible expression.

[0088] The present invention also provides a protein, namely, an α- or β-chain construct, or preferably a TCR receptor construct comprising an α-chain and a β-chain construct, which can specifically bind HLA-*B07:02 to the epitope of SEQ ID NO:2. The protein is preferably encoded by the nucleic acid of the present invention. Preferably, the protein is expressed as a transmembrane protein by a host cell.

[0089] The present invention also provides host cells comprising the nucleic acid or protein of the present invention. The host cells can be eukaryotic cells, such as plants, animals, fungi or algae, or can be prokaryotic cells, such as bacteria or protozoa. The host cells can be cultured cells or primary cells, i.e., directly isolated from an organism (e.g., human). The host cells can be adherent cells or suspension cells, i.e., cells that grow in suspension. For producing recombinant TCR, polypeptide or protein, the host cells are preferably mammalian cells. Most preferably, the host cells are human cells. Although the host cells can be any cell type, can be derived from any type of tissue, and can be at any developmental stage, the host cells are preferably peripheral blood leukocytes (PBLs) or peripheral blood mononuclear cells (PBMCs). More preferably, the host cells are T cells or T cell precursors, especially human T cells. The T cells can be any T cells, such as cultured T cells, e.g., primary T cells, or T cells from a cultured T cell line, or T cells obtained from a mammal, preferably, it is a T cell or T cell precursor from a human patient, especially from a human patient to be treated. T cells of autologous or allogeneic origin can be obtained from various sources, such as blood, bone marrow, lymph nodes, thymus or other tissues or fluids. The T cells can also be enriched or purified. Preferably, the T cells are human T cells. More preferably, the T cells are T cells isolated from humans, such as human patients. The T cells can be any type of T cells, but are preferably CD8+ cells. It can be at any developmental stage, including but not limited to tumor infiltrating cells (TILs), effector cells, central effector cells, memory T cells, naive T cells, etc., preferably central memory T cells.

[0090] The host cells of the present invention preferably comprise the nucleic acid and / or the protein of the present invention, wherein the host cells are preferably CD8+ T cells, optionally, human CD8+ T cells.

[0091] The present invention also provides a pharmaceutical composition comprising

[0092] a) the nucleic acid of the present invention, wherein the nucleic acid encodes a TCR construct capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02; or

[0093] b) the protein of the present invention, wherein the protein comprises a TCR construct capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02; or

[0094] c) the host cells of the present invention, wherein the host cells can express a TCR construct capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02.

[0095] Preferably, the pharmaceutical composition comprises a human CD8+ host cell as defined herein. The host cell can include, for example, a vector encoding a TCR construct, the TCR construct comprising a TCRα-chain construct and a TCRβ-chain construct that are capable of specifically recognizing the peptide of SEQ ID NO:2 in the presence of HLA-B*07:02. Preferably, the vector is an expression vector for expressing the α-chain and β-chain constructs on one nucleic acid, for example, separated by a p2A element. The variable regions of the TCR chains as defined herein, such as TCR2304, TCR2207, TCR2205, TCR1610 or TCR1605, preferably TCR2304, are linked to a constant region, preferably to a minimally murine constant region.

[0096] Optionally, the nucleic acid of the present invention, particularly an expression vector, can also be administered to a patient for in vivo transduction of T cells.

[0097] The pharmaceutical composition can also be part of a kit comprising other therapeutic agents, such as an antibody like rituximab, an immunotoxin (such as inotuzumab ozogamicin), or a CAR that targets a B cell lineage antigen (such as CD19, CD20, CD22 or CD79), preferably a CAR capable of targeting CD19, a small molecule such as a kinase inhibitor or a chemotherapeutic agent, including combination chemotherapy and even high-dose chemotherapy. The pharmaceutical composition can be used in combination with any of the above other therapeutic agents, administered before, concomitantly with, or after the pharmaceutical composition. The pharmaceutical composition of the present invention can also be combined with a reagent capable of inducing IFNγ expression in target tumor cells to enhance the processing of the peptide of ID NO:2 in a composition or kit.

[0098] The pharmaceutical composition of the present invention or the kit of the present invention can be used for the diagnosis, prevention and / or treatment of a disease, particularly in a patient suspected of containing cells expressing a MYD88 protein with an L265P mutation. The disease is preferably a tumor disease, such as a benign or malignant tumor disease. In a preferred embodiment, the tumor cells have been confirmed to express MYD88 L265P and / or HLA-B*07:02, particularly both simultaneously.

[0099] Preferably, the patient has a non-Hodgkin B cell lymphoma selected from the following:

[0100] - Diffuse large B cell lymphoma (DLBCL), preferably activated B cell type DLBCL (ABC-DLBCL) or primary CNS DLBCL, cutaneous DLBCL, leg type DLBCL or testicular DLBCL;

[0101] - Lymphoplasmacytic lymphoma (LPL), preferably Waldenström macroglobulinemia (WM); and

[0102] - IgM monoclonal gammopathy (IgM MGUS).

[0103] Preferably, the disease is treated. A reduction in the risk of disease is also considered prevention of the disease, wherein, preferably, the risk in the treated subject is reduced below normal levels in the comparison population, preferably, the risk is reduced by at least 10%, at least 25%, at least 50%, or at least 75% or 100%.

[0104] The present invention also provides a method of treating a subject having a disease as described above, particularly a tumor or tumor disease, comprising administering a nucleic acid, protein, or host cell of the present invention. Preferably, the subject is a subject in need of such treatment, i.e., a patient. The subject in a preferred embodiment is a mammalian subject having a tumor or tumor disease, preferably a human patient. The active agent is administered in an effective dose.

[0105] A preferred pharmaceutical use of the present invention relates to immunotherapy, preferably adoptive T cell therapy. The products and methods of the present invention are particularly suitable for adoptive T cell therapy. Administration of the compounds of the present invention can include, for example, infusion of the T cells of the present invention into the patient. Preferably, the T cells are autologous T cells of the patient that have been genetically modified to express the TCR of the present invention, for example, they are transduced in vitro with the nucleic acid of the present invention.

[0106] The treatment of the present invention can be a first-line treatment for the patient. Preferably, it is a second-line treatment for the patient if the patient has relapsed or is difficult to cure with one or more alternative agents (e.g., small molecule inhibitors, chemotherapy, antibodies, or CAR-based therapies, such as those directed against B cell lineage antigens like CD19).

[0107] The protein TCR construct of the present invention can also be used, for example, for diagnostic purposes to ascertain whether a subject expresses MYD88 L265P, particularly whether the epitope according to SEQ ID NO:2 is presented by HLA*B07:02. For this purpose, such a construct is preferably labeled to facilitate detection. Preferably, adoptive T cell therapy of the present invention is used for patients found to have the epitope present on HLA*B07:02, or alternatively, TCR gene therapy of the present invention is used for treatment.

[0108] The present invention also provides a method for testing a human subject, such as a patient with B-cell lymphoma expressing MYD88 L265P. The method includes contacting a sample from a subject comprising tumor cells, such as tumor cells derived from the subject's blood, with (preferably labeled) the TCR construct of the present invention, or a host cell having the expression construct of the present invention. The method may further include detecting the label, by FACS or microscopy methods, or detecting the activation of the T cells, which may be FACS-based. Detecting the activation of the T cells may include detecting T cell activation markers such as CD137 (and optionally, the expression of the TCR construct of the present invention), by FACS, detecting the expression of cytokines, for example, by ELISA, ELISPOT or PCR-based methods.

[0109] The method may further include informing the subject of the expression or lack of expression of MYD88 L265P, and optionally, if the patient expresses MYD88 L265P, treating the patient subject with the pharmaceutical composition of the present invention.

[0110] Of course, the presence or absence of MYD88 L265P can also be determined by other means, for example, by sequencing, PCR-based methods or antibody-based methods.

[0111] The present invention also relates to a method for preparing the host cell of the present invention, including introducing an expression vector encoding the TCR construct of the present invention into a suitable host cell, which is preferably a human CD8+ T cell isolated from a patient. Then the host cell can be re-introduced into the patient.

[0112] The present invention will be further illustrated by the following examples in conjunction with the reference drawings and sequences. However, the present invention is not limited thereto. For the purposes of the present invention, all references cited herein are incorporated herein by reference in their entirety. SEQUENCE LISTING <110> Charité - Universitätsmedizin Berlin <120> Specific T cell receptor against the mutant MYD88L265P protein epitope in adoptive T cell therapy <130> 11731 P 6821 EP <150> EP19152801.7 <151> 2019-01-21 <160> 152 <170> BiSSAP 1.3.6 <210> 1 <211> 9 <212> PRT <213> Homo sapiens <220> <223> MYD88 L265P 9mer <400> 1 Arg Pro Ile Pro Ile Lys Tyr Lys Ala 1 5 <210> 2 <211> 10 <212> PRT <213> Homo sapiens <220> <223> MYD88 L265P 10mer <400> 2 Arg Pro Ile Pro Ile Lys Tyr Lys Ala Met 1 5 10 <210> 3 <211> 10 <212> PRT <213> Homo sapiens <220> <223> wt MYD88 10mer <400> 3 Arg Leu Ile Pro Ile Lys Tyr Lys Ala Met 1 5 10 <210> 4 <400> 4 000 <210> 5 <211> 136 <212> PRT <213> Mus musculus <220> <223> Murine constant region TCR alpha chain <400> 5 Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro Arg Ser 1 5 10 15 Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln Ile Asn 20 25 30 Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys Thr Val 35 40 45 Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile Ala Trp 50 55 60 Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu Thr Asn 65 70 75 80 Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu Thr Glu 85 90 95 Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu Ser Val 100 105 110 Met Gly Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn Leu Leu 115 120 125 Met Thr Leu Arg Leu Trp Ser Ser 130 135 <210> 6 <211> 140 <212> PRT <213> Artificial Sequence <220> <223> Mouse Minimal Constant Region - TCR alpha chain <400> 6 Ile Gln Asn Pro Asp Pro Ala Val Tyr Gln Leu Arg Asp Ser Lys Ser 1 5 10 15 Ser Asp Lys Ser Val Cys Leu Phe Thr Asp Phe Asp Ser Gln Thr Asn 20 25 30 Val Ser Gln Ser Lys Asp Ser Asp Val Tyr Ile Thr Asp Lys Thr Val 35 40 45 Leu Asp Met Arg Ser Met Asp Phe Lys Ser Asn Ser Ala Val Ala Trp 50 55 60 Ser Asn Lys Ser Asp Phe Ala Cys Ala Asn Ala Phe Asn Asn Ser Ile 65 70 75 80 Ile Pro Glu Asp Thr Phe Phe Pro Ser Ser Asp Val Pro Cys Asp Val 85 90 95 Lys Leu Val Glu Lys Ser Phe Glu Thr Asp Thr Asn Leu Asn Phe Gln 100 105 110 Asn Leu Ser Val Ile Gly Phe Arg Ile Leu Leu Leu Lys Val Ala Gly 115 120 125 Phe Asn Leu Leu Met Thr Leu Arg Leu Trp Ser Ser 130 135 140 <210> 7 <211> 140 <212> PRT <213> Homo sapiens <220> <223> Human constant region - TCR alpha chain <400> 7 Ile Gln Asn Pro Asp Pro Ala Val Tyr Gln Leu Arg Asp Ser Lys Ser 1 5 10 15 Ser Asp Lys Ser Val Cys Leu Phe Thr Asp Phe Asp Ser Gln Thr Asn 20 25 30 Val Ser Gln Ser Lys Asp Ser Asp Val Tyr Ile Thr Asp Lys Thr Val 35 40 45 Leu Asp Met Arg Ser Met Asp Phe Lys Ser Asn Ser Ala Val Ala Trp 50 55 60 Ser Asn Lys Ser Asp Phe Ala Cys Ala Asn Ala Phe Asn Asn Ser Ile 65 70 75 80 Ile Pro Glu Asp Thr Phe Phe Pro Ser Pro Glu Ser Ser Cys Asp Val 85 90 95 Lys Leu Val Glu Lys Ser Phe Glu Thr Asp Thr Asn Leu Asn Phe Gln 100 105 110 Asn Leu Ser Val Ile Gly Phe Arg Ile Leu Leu Leu Lys Val Ala Gly 115 120 125 Phe Asn Leu Leu Met Thr Leu Arg Leu Trp Ser Ser 130 135 140 <210> 8 <211> 173 <212> PRT <213> Mus musculus <220> <223> Murine constant region - TCR beta chain mTCRBC2 <400> 8 Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu Phe Glu Pro 1 5 10 15 Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu Val Cys Leu 20 25 30 Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp Trp Val Asn 35 40 45 Gly Lys Glu Val His Ser Gly Val Ser Thr Asp Pro Gln Ala Tyr Lys 50 55 60 Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg Val Ser Ala 65 70 75 80 Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln Val Gln Phe 85 90 95 His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser Pro Lys Pro 100 105 110 Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala Asp Cys Gly 115 120 125 Ile Thr Ser Ala Ser Tyr His Gln Gly Val Leu Ser Ala Thr Ile Leu 130 135 140 Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val Leu Val Ser 145 150 155 160 Gly Leu Val Leu Met Ala Met Val Lys Lys Lys Asn Ser 165 170 <210> 9 <211> 179 <212> PRT <213> Artificial Sequence <220> <223> Mouse Minimal Constant Region - TCR beta chain <400> 9 Glu Asp Leu Lys Asn Val Phe Pro Pro Lys Val Ala Val Phe Glu Pro 1 5 10 15 Ser Lys Ala Glu Ile Ala His Thr Gln Lys Ala Thr Leu Val Cys Leu 20 25 30 Ala Thr Gly Phe Tyr Pro Asp His Val Glu Leu Ser Trp Trp Val Asn 35 40 45 Gly Lys Glu Val His Ser Gly Val Ser Thr Asp Pro Gln Pro Leu Lys 50 55 60 Glu Gln Pro Ala Leu Asn Asp Ser Arg Tyr Cys Leu Ser Ser Arg Leu 65 70 75 80 Arg Val Ser Ala Thr Phe Trp Gln Asn Pro Arg Asn His Phe Arg Cys 85 90 95 Gln Val Gln Phe Tyr Gly Leu Ser Glu Asn Asp Glu Trp Thr Gln Asp 100 105 110 Arg Ala Lys Pro Val Thr Gln Ile Val Ser Ala Glu Ala Trp Gly Arg 115 120 125 Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr His Gln Gly Val Leu Ser 130 135 140 Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala 145 150 155 160 Val Leu Val Ser Ala Leu Val Leu Met Ala Met Val Lys Arg Lys Asp 165 170 175 Ser Arg Gly <210> 10 <211> 179 <212> PRT <213> Homo sapiens <220> <223> Human constant region - TCR beta chain TCRBC2 <400> 10 Glu Asp Leu Lys Asn Val Phe Pro Pro Lys Val Ala Val Phe Glu Pro 1 5 10 15 Ser Glu Ala Glu Ile Ser His Thr Gln Lys Ala Thr Leu Val Cys Leu 20 25 30 Ala Thr Gly Phe Tyr Pro Asp His Val Glu Leu Ser Trp Trp Val Asn 35 40 45 Gly Lys Glu Val His Ser Gly Val Ser Thr Asp Pro Gln Pro Leu Lys 50 55 60 Glu Gln Pro Ala Leu Asn Asp Ser Arg Tyr Cys Leu Ser Ser Arg Leu 65 70 75 80 Arg Val Ser Ala Thr Phe Trp Gln Asn Pro Arg Asn His Phe Arg Cys 85 90 95 Gln Val Gln Phe Tyr Gly Leu Ser Glu Asn Asp Glu Trp Thr Gln Asp 100 105 110 Arg Ala Lys Pro Val Thr Gln Ile Val Ser Ala Glu Ala Trp Gly Arg 115 120 125 Ala Asp Cys Gly Phe Thr Ser Glu Ser Tyr Gln Gln Gly Val Leu Ser 130 135 140 Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala 145 150 155 160 Val Leu Val Ser Ala Leu Val Leu Met Ala Met Val Lys Arg Lys Asp 165 170 175 Ser Arg Gly <210> 11 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2207 <400> 11 Thr Ser Gly Phe Asn Gly 1 5 <210> 12 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2207 <400> 12 Asn Val Leu Asp Gly Leu 1 5 <210> 13 <211> 12 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2207 <400> 13 Cys Ala Val Asp Val Gly Tyr Ser Thr Leu Thr Phe 1 5 10 <210> 14 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2207 <400> 14 Asp Phe Gln Ala Thr Thr 1 5 <210> 15 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2207 <400> 15 Ser Asn Glu Gly Ser Lys Ala 1 5 <210> 16 <211> 16 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2207 <400> 16 Cys Ser Ala Arg Asp Arg Ser Gly Thr Leu Gly Gly Glu Leu Phe Phe 1 5 10 15 <210> 17 <211> 127 <212> PRT <213> Homo sapiens <220> <223> Variable region of the alpha chain of TCR2207 <400> 17 Met Trp Gly Val Phe Leu Leu Tyr Val Ser Met Lys Met Gly Gly Thr 1 5 10 15 Thr Gly Gln Asn Ile Asp Gln Pro Thr Glu Met Thr Ala Thr Glu Gly 20 25 30 Ala Ile Val Gln Ile Asn Cys Thr Tyr Gln Thr Ser Gly Phe Asn Gly 35 40 45 Leu Phe Trp Tyr Gln Gln His Ala Gly Glu Ala Pro Thr Phe Leu Ser 50 55 60 Tyr Asn Val Leu Asp Gly Leu Glu Glu Lys Gly Arg Phe Ser Ser Phe 65 70 75 80 Leu Ser Arg Ser Lys Gly Tyr Ser Tyr Leu Leu Leu Lys Glu Leu Gln 85 90 95 Met Lys Asp Ser Ala Ser Tyr Leu Cys Ala Val Asp Val Gly Tyr Ser 100 105 110 Thr Leu Thr Phe Gly Lys Gly Thr Met Leu Leu Val Ser Pro Asp 115 120 125 <210> 18 <211> 132 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2207 <400> 18 Met Leu Leu Leu Leu Leu Leu Leu Gly Pro Gly Ser Gly Leu Gly Ala 1 5 10 15 Val Val Ser Gln His Pro Ser Trp Val Ile Cys Lys Ser Gly Thr Ser 20 25 30 Val Lys Ile Glu Cys Arg Ser Leu Asp Phe Gln Ala Thr Thr Met Phe 35 40 45 Trp Tyr Arg Gln Phe Pro Lys Gln Ser Leu Met Leu Met Ala Thr Ser 50 55 60 Asn Glu Gly Ser Lys Ala Thr Tyr Glu Gln Gly Val Glu Lys Asp Lys 65 70 75 80 Phe Leu Ile Asn His Ala Ser Leu Thr Leu Ser Thr Leu Thr Val Thr 85 90 95 Ser Ala His Pro Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Asp 100 105 110 Arg Ser Gly Thr Leu Gly Gly Glu Leu Phe Phe Gly Glu Gly Ser Arg 115 120 125 Leu Thr Val Leu 130 <210> 19 <211> 381 <212> DNA <213> Artificial sequence <220> <223> Variable region of the alpha chain of TCR2207 - codon optimized <400> 19 atgtggggcg tgttcctgct gtacgtgtcc atgaagatgg gcggcaccac cggccagaac 60 atcgatcagc ctacagagat gaccgccacc gagggcgcca tcgtgcagat caattgcacc 120 taccagacca gcggcttcaa cgggctgttt tggtatcagc agcacgccgg cgaggcccct 180 acattcctga gctacaatgt gctggacggc ctcgaggaaa agggcagatt ctccagcttc 240 ctgagcagaa gcaagggcta ctcctacctg ctgctgaaag aactgcagat gaaggacagc 300 gcctcttacc tgtgcgccgt ggatgtgggc tacagcacac tgacatttgg caagggcacc 360 atgctgctcg tgtccccaga c 381 <210> 20 <211> 396 <212> DNA <213> Artificial sequence <220> <223> Variable region of the beta chain of TCR2207 - codon optimized <400> 20 atgttgttgt tgctgttgct cctcggacct ggctctggac tgggagctgt ggtttctcag 60 cacccctctt gggtcatctg caagagcggc accagcgtga agatcgagtg cagaagcctg 120 gacttccagg ccaccacaat gttctggtac agacagttcc ccaagcagag cctgatgctg 180 atggccacct ctaacgaggg cagcaaggcc acatatgagc agggcgtcga gaaggacaag 240 ttcctgatca accacgccag cctgacactg agcaccctga cagtgacaag cgcccatcct 300 gaggacagca gcttctacat ctgcagcgcc agagacagaa gcggcacact tggcggcgag 360 ctgttttttg gcgagggctc tagactgacc gtgctg 396 <210> 21 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2205 <400> 21 Asn Ile Ala Thr Asn Asp Tyr 1 5 <210> 22 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2205 <400> 22 Gly Tyr Lys Thr Lys 1 5 <210> 23 <211> 14 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2205 <400> 23 Cys Leu Val Gly Arg Asp Gly Gly Ser Tyr Ile Pro Thr Phe 1 5 10 <210> 24 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2205 <400> 24 Ser Gly Asp Leu Ser 1 5 <210> 25 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2205 <400> 25 Tyr Tyr Asn Gly Glu Glu 1 5 <210> 26 <211> 14 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2205 <400> 26 Cys Ala Ser Ser Ala Gly Gln Gly Ala Tyr Glu Gln Tyr Phe 1 5 10 <210> 27 <211> 129 <212> PRT <213> Homo sapiens <220> <223> Variable region of the alpha chain of TCR2205 <400> 27 Met Arg Gln Val Ala Arg Val Ile Val Phe Leu Thr Leu Ser Thr Leu 1 5 10 15 Ser Leu Ala Lys Thr Thr Gln Pro Ile Ser Met Asp Ser Tyr Glu Gly 20 25 30 Gln Glu Val Asn Ile Thr Cys Ser His Asn Asn Ile Ala Thr Asn Asp 35 40 45 Tyr Ile Thr Trp Tyr Gln Gln Phe Pro Ser Gln Gly Pro Arg Phe Ile 50 55 60 Ile Gln Gly Tyr Lys Thr Lys Val Thr Asn Glu Val Ala Ser Leu Phe 65 70 75 80 Ile Pro Ala Asp Arg Lys Ser Ser Thr Leu Ser Leu Pro Arg Val Ser 85 90 95 Leu Ser Asp Thr Ala Val Tyr Tyr Cys Leu Val Gly Arg Asp Gly Gly 100 105 110 Ser Tyr Ile Pro Thr Phe Gly Arg Gly Thr Ser Leu Ile Val His Pro 115 120 125 Tyr <210> 28 <211> 132 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2205 <400> 28 Met Gly Phe Arg Leu Leu Cys Cys Val Ala Phe Cys Leu Leu Gly Ala 1 5 10 15 Gly Pro Val Asp Ser Gly Val Thr Gln Thr Pro Lys His Leu Ile Thr 20 25 30 Ala Thr Gly Gln Arg Val Thr Leu Arg Cys Ser Pro Arg Ser Gly Asp 35 40 45 Leu Ser Val Tyr Trp Tyr Gln Gln Ser Leu Asp Gln Gly Leu Gln Phe 50 55 60 Leu Ile His Tyr Tyr Asn Gly Glu Glu Arg Ala Lys Gly Asn Ile Leu 65 70 75 80 Glu Arg Phe Ser Ala Gln Gln Phe Pro Asp Leu His Ser Glu Leu Asn 85 90 95 Leu Ser Ser Leu Glu Leu Gly Asp Ser Ala Leu Tyr Phe Cys Ala Ser 100 105 110 Ser Ala Gly Gln Gly Ala Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg 115 120 125 Leu Thr Val Thr 130 <210> 29 <211> 387 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2205 - codon optimized <400> 29 atgagacagg tggccagagt gatcgtgttc ctgacactga gcaccctgag cctggccaag 60 Met-Arg-Arg Trp-Pro-Ser Asp-Arg-Val-Phe Leu-Thr-Thr-Glu His-Pro-Glu Pro-Gly-Gln accacacagc ccatcagcat ggacagctac gagggccaag aagtgaacat cacctgtagc 120 Thr-His-Ser Pro-Ser-His Trp-Thr-Leu-Tyr Glu-Gly-Gln Lys-Val-Asn-Ile Thr-Cys-Ser cacaacaata tcgccaccaa cgactacatc acgtggtatc aacagttccc cagtcaaggc 180 His-Gln-Asn-Ile Ala-Thr-Asn Asp-Tyr-Ile Thr-Trp-Tyr Asn-Ser-Phe Pro-Ser-Lys-Ala cctcggttca tcatccaagg ctacaagacc aaagtgacca acgaggtggc ctctctgttc 240 Pro-Arg-Phe Ile-Ile-Pro-Lys Ala-Tyr-Lys-Thr Lys-Val-Thr-Asn Glu-Val-Ala-Ser-Phe atccccgccg acagaaagag cagcaccctg tctctgccta gagtgtccct gagcgatacc 300 Ile-Pro-Ala-Asp Glu-Lys-Glu Gln-His-Leu Ser-Ser-Ala-Leu Ser-Val-Pro Glu-Ala-Tyr gccgtgtact actgtctcgt gggcagagat ggcggcagct acatccctac atttggcaga 360 Ala-Val-Tyr Tyr-Cys-Ser-Val Gly-Gln-Asp Gly-Gly-Ser Tyr-Ile-Pro-Tyr Phe-Gly-Gln ggcacaagcc tgatcgtgca cccctac 387 Gly-His-Lys-Pro Asp-Arg-Cys Thr-Pro-Tyr 387 <210> 30 <211> 396 <212> DNA <213> Artificial Sequence <220> <223> Variable region of beta chain of TCR2205 - codon optimized <400> 30 atgggtttta gactgctgtg ctgcgtggcc ttctgtctgc ttggagctgg ccctgtggat 60 Met-Gly-Phe Arg-Leu-Leu-Cys Leu-Arg-Trp-Ala Phe-Cys-Ser-Leu Glu-Leu-Ala-Leu-Trp-Asp agcggcgtta cccagacacc taagcacctg atcacagcca caggccagcg cgtgaccctg 120 Ser-Gly-Val Tyr-Pro-Asp-Thr Leu-Lys-His-Leu Ile-Thr-Ala Thr-Gly-Gln-Ala Val-Thr-Leu agatgttctc ctagaagcgg cgacctgagc gtgtactggt atcagcagtc tctggaccag 180 Arg-Met-Phe-Ser Leu-Arg-Lys-Gly Asp-Leu-Ser Val-Tyr-Trp Ile-Ser-Ser Ser-Trp-Thr-Gln ggcctgcagt tcctgatcca ctactacaac ggcgaggaaa gagccaaggg caacatcctg 240 gaacggttca gcgcccagca gttcccagat ctgcacagcg agctgaacct gagcagcctg 300 gaactgggag atagcgccct gtacttctgt gcctcttctg ctggacaggg cgcctacgag 360 cagtattttg gccctggcac cagactgacc gtgacc 396 <210> 31 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR1610 <400> 31 Asp Ser Ala Ile Tyr Asn 1 5 <210> 32 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR1610 <400> 32 Ile Gln Ser Ser Gln Arg Glu 1 5 <210> 33 <211> 13 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR1610 <400> 33 Cys Ala Pro Leu Gly Gly Gly Tyr Asn Lys Leu Ile Phe 1 5 10 <210> 34 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR1610 <400> 34 Met Asn His Asn Tyr 1 5 <210> 35 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR1610 <400> 35 Ser Val Gly Ala Gly Ile 1 5 <210> 36 <211> 14 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR1610 <400> 36 Cys Ala Ser Arg Leu Pro Thr Thr Asp Glu Lys Leu Phe Phe 1 5 10 <210> 37 <211> 131 <212> PRT <213> Homo sapiens <220> <223> Variable region of alpha chain of TCR1610 <400> 37 Met Glu Thr Leu Leu Gly Leu Leu Ile Leu Trp Leu Gln Leu Gln Trp 1 5 10 15 Val Ser Ser Lys Gln Glu Val Thr Gln Ile Pro Ala Ala Leu Ser Val 20 25 30 Pro Glu Gly Glu Asn Leu Val Leu Asn Cys Ser Phe Thr Asp Ser Ala 35 40 45 Ile Tyr Asn Leu Gln Trp Phe Arg Gln Asp Pro Gly Lys Gly Leu Thr 50 55 60 Ser Leu Leu Leu Ile Gln Ser Ser Gln Arg Glu Gln Thr Ser Gly Arg 65 70 75 80 Leu Asn Ala Ser Leu Asp Lys Ser Ser Gly Arg Ser Thr Leu Tyr Ile 85 90 95 Ala Ala Ser Gln Pro Gly Asp Ser Ala Thr Tyr Leu Cys Ala Pro Leu 100 105 110 Gly Gly Gly Tyr Asn Lys Leu Ile Phe Gly Ala Gly Thr Arg Leu Ala 115 120 125 Val His Pro 130 <210> 38 <211> 132 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR1610 <400> 38 Met Ser Ile Ser Leu Leu Cys Cys Ala Ala Phe Pro Leu Leu Trp Ala 1 5 10 15 Gly Pro Val Asn Ala Gly Val Thr Gln Thr Pro Lys Phe Arg Ile Leu 20 25 30 Lys Ile Gly Gln Ser Met Thr Leu Gln Cys Ala Gln Asp Met Asn His 35 40 45 Asn Tyr Met Tyr Trp Tyr Arg Gln Asp Pro Gly Met Gly Leu Lys Leu 50 55 60 Ile Tyr Tyr Ser Val Gly Ala Gly Ile Thr Asp Lys Gly Glu Val Pro 65 70 75 80 Asn Gly Tyr Asn Val Ser Arg Ser Thr Thr Glu Asp Phe Pro Leu Arg 85 90 95 Leu Glu Leu Ala Ala Pro Ser Gln Thr Ser Val Tyr Phe Cys Ala Ser 100 105 110 Arg Leu Pro Thr Thr Asp Glu Lys Leu Phe Phe Gly Ser Gly Thr Gln 115 120 125 Leu Ser Val Leu 130 <210> 39 <211> 393 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR1610 - Codon-optimized <400> 39 atggaaacac tgctgggcct gctgatcctg tggctgcaac tgcaatgggt gtcctccaag 60 caagaagtga ctcagatccc tgccgctctg tccgtgcctg aaggcgaaaa cctggtcctg 120 aactgcagct tcaccgacag cgccatctac aacctgcagt ggttcaggca ggatccaggc 180 aagggactga cctctctgct gctgattcag agcagccaga gagagcagac ctccggcaga 240 ctgaatgcca gcctggataa gagcagcggc cggtctacac tgtatatcgc cgcttctcag 300 ccaggcgata gcgccacata tctgtgtgct cctctcggcg gaggctacaa caagctgatt 360 ttcggcgctg gcaccagact ggccgtgcat cct 393 <210> 40 <211> 396 <212> DNA <213> Artificial Sequence <220> <223> Variable region of beta chain of TCR1610 - codon optimized <400> 40 atgtctatta gcctgctgtg ctgtgccgcc tttcctctgc tttgggccgg acctgttaat 60 gccggcgtga cccagacacc taagttccgg atcctgaaga tcggccagag catgaccctg 120 cagtgcgccc aggacatgaa ccacaactac atgtactggt acagacagga ccccggcatg 180 ggcctgaagc tgatctacta ttctgtcgga gccggcatca ccgacaaggg cgaagtgcct 240 aatggctaca acgtgtccag aagcaccacc gaggacttcc ctctgcgact ggaactggct 300 gccccatctc agaccagcgt gtacttctgt gccagcagac tgcccaccac cgacgagaag 360 ctgttttttg gcagcggcac ccagctgagc gtgctg 396 <210> 41 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR1605 <400> 41 Asn Ser Ala Ser Asp Tyr 1 5 <210> 42 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR1605 <400> 42 Ile Arg Ser Asn Met Asp Lys 1 5 <210> 43 <211> 13 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR1605 <400> 43 Cys Ala Glu Gly Thr Gly Ser Ala Arg Gln Leu Thr Phe 1 5 10 <210> 44 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR1605 <400> 44 Met Asp His Glu Asn 1 5 <210> 45 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR1605 <400> 45 Ser Tyr Asp Val Lys Met 1 5 <210> 46 <211> 20 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR1605 <400> 46 Cys Ala Ser Gly Pro Phe Arg Asp Ser Val Leu Thr Leu Val Ala Asn 1 5 10 15 Val Leu Thr Phe 20 <210> 47 <211> 132 <212> PRT <213> Homo sapiens <220> <223> Variable region of alpha chain of TCR1605 <400> 47 Met Ala Gly Ile Arg Ala Leu Phe Met Tyr Leu Trp Leu Gln Leu Asp 1 5 10 15 Trp Val Ser Arg Gly Glu Ser Val Gly Leu His Leu Pro Thr Leu Ser 20 25 30 Val Gln Glu Gly Asp Asn Ser Ile Ile Asn Cys Ala Tyr Ser Asn Ser 35 40 45 Ala Ser Asp Tyr Phe Ile Trp Tyr Lys Gln Glu Ser Gly Lys Gly Pro 50 55 60 Gln Phe Ile Ile Asp Ile Arg Ser Asn Met Asp Lys Arg Gln Gly Gln 65 70 75 80 Arg Val Thr Val Leu Leu Asn Lys Thr Val Lys His Leu Ser Leu Gln 85 90 95 Ile Ala Ala Thr Gln Pro Gly Asp Ser Ala Val Tyr Phe Cys Ala Glu 100 105 110 Gly Thr Gly Ser Ala Arg Gln Leu Thr Phe Gly Ser Gly Thr Gln Leu 115 120 125 Thr Val Leu Pro 130 <210> 48 <211> 138 <212> PRT <213> Homo sapiens <220> <223> Variable region of beta chain of TCR1605 <400> 48 Met Gly Ile Arg Leu Leu Cys Arg Val Ala Phe Cys Phe Leu Ala Val 1 5 10 15 Gly Leu Val Asp Val Lys Val Thr Gln Ser Ser Arg Tyr Leu Val Lys 20 25 30 Arg Thr Gly Glu Lys Val Phe Leu Glu Cys Val Gln Asp Met Asp His 35 40 45 Glu Asn Met Phe Trp Tyr Arg Gln Asp Pro Gly Leu Gly Leu Arg Leu 50 55 60 Ile Tyr Phe Ser Tyr Asp Val Lys Met Lys Glu Lys Gly Asp Ile Pro 65 70 75 80 Glu Gly Tyr Ser Val Ser Arg Glu Lys Lys Glu Arg Phe Ser Leu Ile 85 90 95 Leu Glu Ser Ala Ser Thr Asn Gln Thr Ser Met Tyr Leu Cys Ala Ser 100 105 110 Gly Pro Phe Arg Asp Ser Val Leu Thr Leu Val Ala Asn Val Leu Thr 115 120 125 Phe Gly Ala Gly Ser Arg Leu Thr Val Leu 130 135 <210> 49 <211> 396 <212> DNA <213> Artificial Sequence <220> <223> Variable region of the alpha chain of TCR1605 - codon optimized <400> 49 atggccggaa tcagagccct gttcatgtat ctgtggctgc agctggactg ggtgtccagg 60 ggagaatctg tcggactgca tctgcccaca ctgagcgtgc aagagggcga caacagcatc 120 atcaactgcg cctacagcaa cagcgcctcc gactacttca tctggtacaa gcaagagagc 180 ggcaagggcc ctcagttcat catcgacatc cggtccaaca tggacaagcg gcaaggccag 240 agagtgaccg tcctgctgaa caagaccgtg aagcacctga gcctgcagat cgccgctaca 300 cagcctggcg atagcgccgt gtacttttgt gctgaaggca ccggaagcgc cagacagctg 360 acatttggca gcggaaccca gctcacagtg ctgccc 396 <210> 50 <211> 414 <212> DNA <213> Artificial Sequence <220> <223> Variable region of beta chain of TCR1605 - codon optimized <400> 50 atgggaatta gactgctgtg cagagtggcc ttctgcttcc tggctgttgg cctggtggac 60 gtgaaagtga cccagagcag cagatacctg gtcaagagaa ccggcgagaa ggtgttcctg 120 gaatgcgtgc aggacatgga ccacgagaat atgttctggt acagacagga ccccggcctg 180 ggcctgagac tgatctactt cagctacgac gtgaagatga aggaaaaggg cgacatcccc 240 gagggctaca gcgtgtccag agagaagaaa gagcggttca gcctgatcct ggaaagcgcc 300 agcaccaacc agaccagcat gtacctgtgt gccagcggac ccttcagaga cagcgtgctg 360 acactggtgg ccaacgtgct gacttttggc gccggaagca gactgaccgt gctg 414 <210> 51 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2202 <400> 51 Asn Ile Ala Thr Asn Asp Tyr 1 5 <210> 52 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2202 <400> 52 Gly Tyr Glu Thr Lys 1 5 <210> 53 <211> 13 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2202 <400> 53 Cys Leu Ser Leu Ser Asp Ser Asn Tyr Gln Leu Ile Trp 1 5 10 <210> 54 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2202 <400> 54 Ser Gly Asp Leu Ser 1 5 <210> 55 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2202 <400> 55 Tyr Tyr Asn Gly Glu Glu 1 5 <210> 56 <211> 14 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR 2202 <400> 56 Cys Ala Ser Ser Val Gly Gln Gly Ser Tyr Glu Gln Tyr Phe 1 5 10 <210> 57 <211> 128 <212> PRT <213> Homo sapiens <220> <223> Variable region of the alpha chain of TCR2202 <400> 57 Met Arg Gln Val Ala Arg Val Ile Val Phe Leu Thr Leu Ser Thr Leu 1 5 10 15 Ser Leu Ala Lys Thr Thr Gln Pro Ile Ser Met Asp Ser Tyr Glu Gly 20 25 30 Gln Glu Val Asn Ile Thr Cys Ser His Asn Asn Ile Ala Thr Asn Asp 35 40 45 Tyr Ile Thr Trp Tyr Gln Gln Phe Pro Ser Gln Gly Pro Arg Phe Ile 50 55 60 Ile Gln Gly Tyr Lys Thr Lys Val Thr Asn Glu Val Ala Ser Leu Phe 65 70 75 80 Ile Pro Ala Asp Arg Lys Ser Ser Thr Leu Ser Leu Pro Arg Val Ser 85 90 95 Leu Ser Asp Thr Ala Val Tyr Tyr Cys Leu Ser Leu Ser Asp Ser Asn 100 105 110 Tyr Gln Leu Ile Trp Gly Ala Gly Thr Lys Leu Ile Ile Lys Pro Asp 115 120 125 <210> 58 <211> 132 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2202 <400> 58 Met Gly Phe Arg Leu Leu Cys Cys Val Ala Phe Cys Leu Leu Gly Ala 1 5 10 15 Gly Pro Val Asp Ser Gly Val Thr Gln Thr Pro Lys His Leu Ile Thr 20 25 30 Ala Thr Gly Gln Arg Val Thr Leu Arg Cys Ser Pro Arg Ser Gly Asp 35 40 45 Leu Ser Val Tyr Trp Tyr Gln Gln Ser Leu Asp Gln Gly Leu Gln Phe 50 55 60 Leu Ile Gln Tyr Tyr Asn Gly Glu Glu Arg Ala Lys Gly Asn Ile Leu 65 70 75 80 Glu Arg Phe Ser Ala Gln Gln Phe Pro Asp Leu His Ser Glu Leu Asn 85 90 95 Leu Ser Ser Leu Glu Leu Gly Asp Ser Ala Leu Tyr Phe Cys Ala Ser 100 105 110 Ser Val Gly Gln Gly Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg 115 120 125 Leu Thr Val Thr 130 <210> 59 <211> 384 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2202 - Codon optimized <400> 59 atgagacagg tggccagagt gatcgtgttc ctgacactga gcaccctgag cctggccaag 60 accacacagc ccatcagcat ggactcctac gagggccaag aagtgaacat cacctgtagc 120 cacaacaata tcgccaccaa cgactacatc acgtggtatc aacagttccc cagtcaaggc 180 cctcggttca tcatccaagg ctacaagacc aaagtgacca acgaggtggc ctctctgttc 240 atccccgccg atagaaagag cagcaccctg tctctgccca gagtgtccct gagcgatacc 300 gccgtgtact actgtctgag cctgtccgac tccaactacc agctgatttg gggagccggc 360 accaagctga tcatcaagcc cgac 384 <210> 60 <211> 396 <212> PRT <213> Artificial Sequence <220> <223> Variable region of beta chain of TCR2202 - codon optimized <400> 60 Ala Thr Gly Gly Gly Thr Thr Thr Thr Ala Gly Ala Cys Thr Gly Cys 1 5 10 15 Thr Gly Thr Gly Cys Thr Gly Cys Gly Thr Gly Gly Cys Cys Thr Thr 20 25 30 Cys Thr Gly Thr Cys Thr Gly Cys Thr Thr Gly Gly Ala Gly Cys Thr 35 40 45 Gly Gly Cys Cys Cys Thr Gly Thr Gly Gly Ala Thr Ala Gly Cys Gly 50 55 60 Gly Cys Gly Thr Thr Ala Cys Cys Cys Ala Gly Ala Cys Ala Cys Cys 65 70 75 80 Thr Ala Ala Gly Cys Ala Cys Cys Thr Gly Ala Thr Cys Ala Cys Ala 85 90 95 Gly Cys Cys Ala Cys Ala Gly Gly Cys Cys Ala Gly Cys Gly Cys Gly 100 105 110 Thr Gly Ala Cys Cys Cys Thr Gly Ala Gly Ala Thr Gly Thr Thr Cys 115 120 125 Thr Cys Cys Thr Ala Gly Ala Ala Gly Cys Gly Gly Cys Gly Ala Cys 130 135 140 Cys Thr Gly Ala Gly Cys Gly Thr Gly Thr Ala Cys Thr Gly Gly Thr 145 150 155 160 Ala Thr Cys Ala Gly Cys Ala Gly Thr Cys Thr Cys Thr Gly Gly Ala 165 170 175 Cys Cys Ala Gly Gly Gly Cys Cys Thr Gly Cys Ala Gly Thr Thr Cys 180 185 190 Cys Thr Gly Ala Thr Cys Cys Ala Gly Thr Ala Cys Thr Ala Cys Ala 195 200 205 Ala Cys Gly Gly Cys Gly Ala Gly Gly Ala Ala Ala Gly Ala Gly Cys 210 215 220 Cys Ala Ala Gly Gly Gly Cys Ala Ala Cys Ala Thr Cys Cys Thr Gly 225 230 235 240 Gly Ala Ala Cys Gly Gly Thr Thr Cys Ala Gly Cys Gly Cys Cys Cys 245 250 255 Ala Gly Cys Ala Gly Thr Thr Cys Cys Cys Ala Gly Ala Thr Cys Thr 260 265 270 Gly Cys Ala Cys Ala Gly Cys Gly Ala Gly Cys Thr Gly Ala Ala Cys 275 280 285 Cys Thr Gly Ala Gly Cys Ala Gly Cys Cys Thr Gly Gly Ala Ala Cys 290 295 300 Thr Gly Gly Gly Ala Gly Ala Thr Ala Gly Cys Gly Cys Cys Cys Thr 305 310 315 320 Gly Thr Ala Cys Thr Thr Cys Thr Gly Thr Gly Cys Cys Thr Cys Thr 325 330 335 Thr Cys Thr Gly Thr Cys Gly Gly Cys Cys Ala Gly Gly Gly Cys Ala 340 345 350 Gly Cys Thr Ala Cys Gly Ala Gly Cys Ala Gly Thr Ala Thr Thr Thr 355 360 365 Thr Gly Gly Cys Cys Cys Thr Gly Gly Cys Ala Cys Cys Ala Gly Ala 370 375 380 Cys Thr Gly Ala Cys Cys Gly Thr Gly Ala Cys Cys 385 390 395 <210> 61 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2219 <400> 61 Lys Ala Leu Tyr Ser 1 5 <210> 62 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2219 <400> 62 Leu Leu Lys Gly Gly Glu Gln 1 5 <210> 63 <211> 15 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2219 <400> 63 Cys Gly Thr Ala His Leu Arg Ala Gly Ser Tyr Gln Leu Thr Phe 1 5 10 15 <210> 64 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2219 <400> 64 Met Asn His Glu Tyr 1 5 <210> 65 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2219 <400> 65 Ser Met Asn Val Glu Val 1 5 <210> 66 <211> 16 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2219 <400> 66 Cys Ala Ser Ser Ser Ser Ser Gly Gly Ala Phe Asn Glu Gln Phe Phe 1 5 10 15 <210> 67 <211> 134 <212> PRT <213> Homo sapiens <220> <223> Variable region of the alpha chain of TCR2219 <400> 67 Met Glu Thr Leu Leu Lys Val Leu Ser Gly Thr Leu Leu Trp Gln Leu 1 5 10 15 Thr Trp Val Arg Ser Gln Gln Pro Val Gln Ser Pro Gln Ala Val Ile 20 25 30 Leu Arg Glu Gly Glu Asp Ala Val Ile Asn Cys Ser Ser Ser Lys Ala 35 40 45 Leu Tyr Ser Val His Trp Tyr Arg Gln Lys His Gly Glu Ala Pro Val 50 55 60 Phe Leu Met Ile Leu Leu Lys Gly Gly Glu Gln Met Arg His Glu Lys 65 70 75 80 Ile Phe Ala Ser Phe Asn Glu Lys Lys Gln Gln Ser Ser Leu Tyr Leu 85 90 95 Thr Ala Ser Gln Leu Ser Tyr Ser Gly Thr Tyr Phe Cys Gly Thr Ala 100 105 110 His Leu Arg Ala Gly Ser Tyr Gln Leu Thr Phe Gly Lys Gly Thr Lys 115 120 125 Leu Ser Val Ile Pro Asn 130 <210> 68 <211> 134 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2219 <400> 68 Met Gly Pro Gln Leu Leu Gly Tyr Val Val Leu Cys Leu Leu Gly Ala 1 5 10 15 Gly Pro Leu Glu Ala Gln Val Thr Gln Asn Pro Arg Tyr Leu Ile Thr 20 25 30 Val Thr Gly Lys Lys Leu Thr Val Thr Cys Ser Gln Asn Met Asn His 35 40 45 Glu Tyr Met Ser Trp Tyr Arg Gln Asp Pro Gly Leu Gly Leu Arg Gln 50 55 60 Ile Tyr Tyr Ser Met Asn Val Glu Val Thr Asp Lys Gly Asp Val Pro 65 70 75 80 Glu Gly Tyr Lys Val Ser Arg Lys Glu Lys Arg Asn Phe Pro Leu Ile 85 90 95 Leu Glu Ser Pro Ser Pro Asn Gln Thr Ser Leu Tyr Phe Cys Ala Ser 100 105 110 Ser Ser Ser Ser Gly Gly Ala Phe Asn Glu Gln Phe Phe Gly Pro Gly 115 120 125 Thr Arg Leu Thr Val Leu 130 <210> 69 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2219 - Codon - optimized <400> 69 atggaaaccc tgctgaaggt gctgagcggc acactgctgt ggcagctgac atgggtccga 60 tctcagcagc ctgtgcagtc tcctcaggct gtgatcctga gagaaggcga ggacgccgtg 120 atcaactgca gcagctctaa ggccctgtac agcgtgcact ggtacaggca gaaacacggc 180 gaggcccctg tgttcctgat gattctgctg aaaggcggcg agcagatgcg gcacgagaag 240 atctttgcca gcttcaatga gaagaagcag cagagcagtc tgtacctgac cgccagccag 300 ctgagctaca gcggcacata cttttgcggc acagcccacc tgagagccgg cagctatcag 360 ctgacctttg gcaagggcac aaagctgagc gtgatcccca ac 402 <210> 70 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Variable Region of the beta Chain of TCR2219 - Codon Optimized <400> 70 atgggacctc aactgctggg atatgtggtg ctgtgtctgc tcggagccgg acctctggaa 60 gctcaagtga cacagaaccc cagatacctg atcaccgtga ccggcaagaa actgaccgtg 120 acctgcagcc agaacatgaa ccacgagtac atgagctggt acagacagga ccctggcctg 180 ggcctgagac agatctacta cagcatgaac gtggaagtga ccgacaaggg cgacgtgccc 240 gagggctaca aggtgtccag aaaagagaag cggaacttcc cactgatcct ggaaagccca 300 tctcctaacc agaccagcct gtacttctgc gccagcagca gttctagcgg cggagccttc 360 aacgagcagt tctttggccc tggcaccagg ctgaccgtgc tg 402 <210> 71 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha Chain TCR 1336 <400> 71 Asp Ser Ala Ser Asn Tyr 1 5 <210> 72 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR 1336 <400> 72 Ile Arg Ser Asn Val Gly Glu 1 5 <210> 73 <211> 13 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR 1336 <400> 73 Cys Ala Ala Ser Gly Arg Tyr Asp Tyr Lys Leu Ser Phe 1 5 10 <210> 74 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR 1336 <400> 74 Leu Asn His Asn Val 1 5 <210> 75 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR 1336 <400> 75 Tyr Tyr Asp Lys Asp Phe 1 5 <210> 76 <211> 17 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR 1336 <400> 76 Cys Ala Thr Ala Ser Asp Leu Gln Gly Asp Arg Ser Thr Glu Ala Phe 1 5 10 15 Phe <210> 77 <211> 131 <212> PRT <213> Homo sapiens <220> <223> Variable region of alpha chain of TCR1336 <400> 77 Met Thr Ser Ile Arg Ala Val Phe Ile Phe Leu Trp Leu Gln Leu Asp 1 5 10 15 Leu Val Asn Gly Glu Asn Val Glu Gln His Pro Ser Thr Leu Ser Val 20 25 30 Gln Glu Gly Asp Ser Ala Val Ile Lys Cys Thr Tyr Ser Asp Ser Ala 35 40 45 Ser Asn Tyr Phe Pro Trp Tyr Lys Gln Glu Leu Gly Lys Arg Pro Gln 50 55 60 Leu Ile Ile Asp Ile Arg Ser Asn Val Gly Glu Lys Lys Asp Gln Arg 65 70 75 80 Ile Ala Val Thr Leu Asn Lys Thr Ala Lys His Phe Ser Leu His Ile 85 90 95 Thr Glu Thr Gln Pro Glu Asp Ser Ala Val Tyr Phe Cys Ala Ala Ser 100 105 110 Gly Arg Tyr Asp Tyr Lys Leu Ser Phe Gly Ala Gly Thr Thr Val Thr 115 120 125 Val Arg Ala 130 <210> 78 <211> 135 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR1336 <400> 78 Met Gly Pro Gly Leu Leu His Trp Met Ala Leu Cys Leu Leu Gly Thr 1 5 10 15 Gly His Gly Asp Ala Met Val Ile Gln Asn Pro Arg Tyr Gln Val Thr 20 25 30 Gln Phe Gly Lys Pro Val Thr Leu Ser Cys Ser Gln Thr Leu Asn His 35 40 45 Asn Val Met Tyr Trp Tyr Gln Gln Lys Ser Ser Gln Ala Pro Lys Leu 50 55 60 Leu Phe His Tyr Tyr Asp Lys Asp Phe Asn Asn Glu Ala Asp Thr Pro 65 70 75 80 Asp Asn Phe Gln Ser Arg Arg Pro Asn Thr Ser Phe Cys Phe Leu Asp 85 90 95 Ile Arg Ser Pro Gly Leu Gly Asp Ala Ala Met Tyr Leu Cys Ala Thr 100 105 110 Ala Ser Asp Leu Gln Gly Asp Arg Ser Thr Glu Ala Phe Phe Gly Gln 115 120 125 Gly Thr Arg Leu Thr Val Val 130 135 <210> 79 <211> 393 <212> DNA <213> Synthetic sequence <220> <223> Variable region of the alpha chain of TCR1336 - codon optimized <400> 79 atgaccagca tccgggccgt gttcatcttc ctgtggctgc agctggacct cgtgaacggc 60 gagaatgttg agcagcaccc cagcacactg agcgtgcaag agggcgattc tgccgtgatc 120 aagtgcacct acagcgacag cgcctccaac tacttcccct ggtacaagca agagctggga 180 aaaagacccc agctgatcat cgacatccgg tccaacgtgg gcgagaagaa ggaccagaga 240 atcgccgtga ctctgaacaa gaccgccaag cacttctccc tgcacatcac cgagacacag 300 cctgaggata gcgccgtgta cttttgtgcc gccagcggca gatacgacta caagctgtct 360 tttggcgccg gaaccaccgt gacagtgcgg gcc 393 <210> 80 <211> 405 <212> DNA <213> Synthetic sequence <220> <223> Variable region of beta chain of TCR1336 - codon optimized <400> 80 atgggacctg gattgcttca ttggatggcc ctgtgtctgc tcggcacagg acatggcgac 60 gctatggtca ttcagaaccc cagataccaa gtgacccagt tcggcaagcc cgtgacactg 120 agctgtagcc agacactgaa ccacaacgtg atgtactggt atcagcagaa gtcctctcag 180 gcccctaagc tgctgttcca ctactacgac aaggacttca acaacgaggc cgacacaccc 240 gacaacttcc agagcagaag gcccaatacc agcttctgct tcctggacat cagaagccct 300 ggcctgggag atgccgccat gtatctgtgt gccacagcca gcgatctgca gggcgataga 360 agcaccgagg ccttttttgg ccaaggcacc agactgaccg tggtg 405 <210> 81 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2211 <400> 81 Thr Ile Ser Gly Asn Glu Tyr 1 5 <210> 82 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2211 <400> 82 Gly Leu Lys Asn Asn 1 5 <210> 83 <211> 15 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2211 <400> 83 Cys Ile Val Arg Val Met Lys Thr Ser Tyr Asp Lys Val Ile Phe 1 5 10 15 <210> 84 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2211 <400> 84 Trp Asn His Asn Asn 1 5 <210> 85 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2211 <400> 85 Ser Tyr Gly Val His Asp 1 5 <210> 86 <211> 18 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2211 <400> 86 Cys Ala Ser Ser Glu Pro Arg Thr Ser Gly Ile Ser Tyr Asn Glu Gln 1 5 10 15 Phe Phe <210> 87 <211> 130 <212> PRT <213> Homo sapiens <220> <223> Variable region of the alpha chain of TCR2211 <400> 87 Met Arg Leu Val Ala Arg Val Thr Val Phe Leu Thr Phe Gly Thr Ile 1 5 10 15 Ile Asp Ala Lys Thr Thr Gln Pro Pro Ser Met Asp Cys Ala Glu Gly 20 25 30 Arg Ala Ala Asn Leu Pro Cys Asn His Ser Thr Ile Ser Gly Asn Glu 35 40 45 Tyr Val Tyr Trp Tyr Arg Gln Ile His Ser Gln Gly Pro Gln Tyr Ile 50 55 60 Ile His Gly Leu Lys Asn Asn Glu Thr Asn Glu Met Ala Ser Leu Ile 65 70 75 80 Ile Thr Glu Asp Arg Lys Ser Ser Thr Leu Ile Leu Pro His Ala Thr 85 90 95 Leu Arg Asp Thr Ala Val Tyr Tyr Cys Ile Val Arg Val Met Lys Thr 100 105 110 Ser Tyr Asp Lys Val Ile Phe Gly Pro Gly Thr Ser Leu Ser Val Ile 115 120 125 Pro Asn 130 <210> 88 <211> 136 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2211 <400> 88 Met Gly Thr Arg Leu Phe Phe Tyr Val Ala Leu Cys Leu Leu Trp Ala 1 5 10 15 Gly His Arg Asp Ala Glu Ile Thr Gln Ser Pro Arg His Lys Ile Thr 20 25 30 Glu Thr Gly Arg Gln Val Thr Leu Ala Cys His Gln Thr Trp Asn His 35 40 45 Asn Asn Met Phe Trp Tyr Arg Gln Asp Leu Gly His Gly Leu Arg Leu 50 55 60 Ile His Tyr Ser Tyr Gly Val His Asp Thr Asn Lys Gly Glu Val Ser 65 70 75 80 Asp Gly Tyr Ser Val Ser Arg Ser Asn Thr Glu Asp Leu Pro Leu Thr 85 90 95 Leu Glu Ser Ala Ala Ser Ser Gln Thr Ser Val Tyr Phe Cys Ala Ser 100 105 110 Ser Glu Pro Arg Thr Ser Gly Ile Ser Tyr Asn Glu Gln Phe Phe Gly 115 120 125 Pro Gly Thr Arg Leu Thr Val Leu 130 135 <210> 89 <211> 390 <212> DNA <213> Artificial sequence <220> <223> Variable region of the alpha chain of TCR2211 - codon optimized <400> 89 atgagactgg tggccagagt gacagtgttc ctgaccttcg gcaccatcat cgacgccaag 60 acaacccagc ctcctagcat ggattgtgcc gagggcagag ctgccaacct gccttgtaat 120 cacagcacca tcagcggcaa cgagtacgtg tactggtaca ggcagatcca ctctcagggc 180 cctcagtaca tcatccacgg actgaagaac aacgagacaa acgagatggc cagcctgatc 240 atcaccgagg atagaaagag cagcaccctg atcctgcctc acgccacact gagagatacc 300 gccgtgtact actgcatcgt gcgcgtgatg aagaccagct acgacaaagt gatctttggc 360 cccggaacca gcctgagcgt gatccccaat 390 <210> 90 <211> 408 <212> DNA <213> Artificial sequence <220> <223> Variable region of the beta chain of TCR2211 - codon optimized <400> 90 atgggaacca gactgttttt ttacgtggcc ctgtgcctgc tgtgggccgg acatagagat 60 gccgagatca cacagagccc cagacacaag atcaccgaga caggcagaca agtgaccctg 120 gcctgtcacc agacctggaa ccacaacaac atgttctggt acagacagga cctcggccac 180 ggcctgagac tgatccacta ctcttacggc gtgcacgaca ccaacaaggg cgaagtgtct 240 gacggctaca gcgtgtccag aagcaacacc gaggacctgc ctctgacact ggaatctgcc 300 gccagctctc agaccagcgt gtacttttgt gccagcagcg agcctagaac cagcggcatc 360 agctacaacg agcagttctt cggccctggc accagactga ccgtgctg 408 <210> 91 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2304 <400> 91 Thr Ser Gly Phe Asn Gly 1 5 <210> 92 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2304 <400> 92 Asn Val Leu Asp Gly Leu 1 5 <210> 93 <211> 13 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2304 <400> 93 Cys Ala Val Arg Ala Ser Gly Thr Tyr Lys Tyr Ile Phe 1 5 10 <210> 94 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2304 <400> 94 Ser Gly His Asn Ser 1 5 <210> 95 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2304 <400> 95 Phe Asn Asn Asn Val Pro 1 5 <210> 96 <211> 11 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2304 <400> 96 Cys Ala Ser Gln Asp Ser Tyr Glu Gln Tyr Phe 1 5 10 <210> 97 <211> 128 <212> PRT <213> Homo sapiens <220> <223> Variable region of alpha chain of TCR2304 <400> 97 Met Trp Gly Val Phe Leu Leu Tyr Val Ser Met Lys Met Gly Gly Thr 1 5 10 15 Thr Gly Gln Asn Ile Asp Gln Pro Thr Glu Met Thr Ala Thr Glu Gly 20 25 30 Ala Ile Val Gln Ile Asn Cys Thr Tyr Gln Thr Ser Gly Phe Asn Gly 35 40 45 Leu Phe Trp Tyr Gln Gln His Ala Gly Glu Ala Pro Thr Phe Leu Ser 50 55 60 Tyr Asn Val Leu Asp Gly Leu Glu Glu Lys Gly Arg Phe Ser Ser Phe 65 70 75 80 Leu Ser Arg Ser Lys Gly Tyr Ser Tyr Leu Leu Leu Lys Glu Leu Gln 85 90 95 Met Lys Asp Ser Ala Ser Tyr Leu Cys Ala Val Arg Ala Ser Gly Thr 100 105 110 Tyr Lys Tyr Ile Phe Gly Thr Gly Thr Arg Leu Lys Val Leu Ala Asn 115 120 125 <210> 98 <211> 130 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2304 <400> 98 Met Asp Ser Trp Thr Phe Cys Cys Val Ser Leu Cys Ile Leu Val Ala 1 5 10 15 Lys His Thr Asp Ala Gly Val Ile Gln Ser Pro Arg His Glu Val Thr 20 25 30 Glu Met Gly Gln Glu Val Thr Leu Arg Cys Lys Pro Ile Ser Gly His 35 40 45 Asn Ser Leu Phe Trp Tyr Arg Gln Thr Met Met Arg Gly Leu Glu Leu 50 55 60 Leu Ile Tyr Phe Asn Asn Asn Val Pro Ile Asp Asp Ser Gly Met Pro 65 70 75 80 Glu Asp Arg Phe Ser Ala Lys Met Pro Asn Ala Ser Phe Ser Thr Leu 85 90 95 Lys Ile Gln Pro Ser Glu Pro Arg Asp Ser Ala Val Tyr Phe Cys Ala 100 105 110 Ser Gln Asp Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr 115 120 125 Val Thr 130 <210> 99 <211> 384 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2304 - codon optimized <400> 99 atgtggggcg tgttcctgct gtacgtgtcc atgaagatgg gcggcaccac cggccagaac 60 atcgatcagc ctacagagat gaccgccacc gagggcgcca tcgtgcagat caattgcacc 120 taccagacca gcggcttcaa cgggctgttt tggtatcagc agcacgccgg cgaggcccct 180 acattcctga gctacaatgt gctggacggc ctggaagaaa agggcagatt cagcagcttc 240 ctgagcagaa gcaagggcta ctcctacctg ctgctgaaag aactgcagat gaaggacagc 300 gcctcttacc tgtgtgccgt tagagccagc ggcacctaca agtacatctt cggcaccggc 360 accaggctga aggtgctggc caat 384 <210> 100 <211> 390 <212> DNA <213> Artificial Sequence <220> <223> Variable Region of Beta Chain of TCR2304 - Codon Optimized <400> 100 atggatagct ggaccttttg ttgcgtgtcc ctgtgcatcc tggtggccaa gcacacagat 60 gccggcgtga tccagtctcc tagacacgaa gtgaccgaga tgggccaaga agtgaccctg 120 cgctgcaagc ctatcagcgg ccacaatagc ctgttctggt acagacagac catgatgaga 180 ggcctggaac tgctgatcta cttcaacaac aacgtgccca tcgacgacag cggcatgccc 240 gaggatagat tcagcgccaa gatgcccaac gccagcttca gcaccctgaa gatccagcct 300 agcgagccca gagatagcgc cgtgtacttt tgcgccagcc aggacagcta cgagcagtac 360 tttggccctg gcaccagact gaccgtgacc 390 <210> 101 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2705 <400> 101 Asn Ser Ala Phe Gln Tyr 1 5 <210> 102 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2705 <400> 102 Thr Tyr Ser Ser Gly Asn 1 5 <210> 103 <211> 13 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2705 <400> 103 Cys Ala Met Ser Gly Thr Gly Gly Phe Lys Thr Ile Phe 1 5 10 <210> 104 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2705 <400> 104 Leu Gly His Asn Ala 1 5 <210> 105 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2705 <400> 105 Tyr Ser Leu Glu Glu Arg 1 5 <210> 106 <211> 15 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2705 <400> 106 Cys Ala Ser Ser Gln Asp Arg Pro Asn Tyr Tyr Gly Tyr Thr Phe 1 5 10 15 <210> 107 <211> 134 <212> PRT <213> Homo sapiens <220> <223> Variable region of alpha chain of TCR2705 <400> 107 Met Met Lys Ser Leu Arg Val Leu Leu Val Ile Leu Trp Leu Gln Leu 1 5 10 15 Ser Trp Val Trp Ser Gln Gln Lys Glu Val Glu Gln Asp Pro Gly Pro 20 25 30 Leu Ser Val Pro Glu Gly Ala Ile Val Ser Leu Asn Cys Thr Tyr Ser 35 40 45 Asn Ser Ala Phe Gln Tyr Phe Met Trp Tyr Arg Gln Tyr Ser Arg Lys 50 55 60 Gly Pro Glu Leu Leu Met Tyr Thr Tyr Ser Ser Gly Asn Lys Glu Asp 65 70 75 80 Gly Arg Phe Thr Ala Gln Val Asp Lys Ser Ser Lys Tyr Ile Ser Leu 85 90 95 Phe Ile Arg Asp Ser Gln Pro Ser Asp Ser Ala Thr Tyr Leu Cys Ala 100 105 110 Met Ser Gly Thr Gly Gly Phe Lys Thr Ile Phe Gly Ala Gly Thr Arg 115 120 125 Leu Phe Val Lys Ala Asn 130 <210> 108 <211> 136 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2705 <400> 108 Met Gly Cys Arg Leu Leu Cys Cys Ala Val Leu Cys Leu Leu Gly Ala 1 5 10 15 Gly Glu Leu Val Pro Met Glu Thr Gly Val Thr Gln Thr Pro Arg His 20 25 30 Leu Val Met Gly Met Thr Asn Lys Lys Ser Leu Lys Cys Glu Gln His 35 40 45 Leu Gly His Asn Ala Met Tyr Trp Tyr Lys Gln Ser Ala Lys Lys Pro 50 55 60 Leu Glu Leu Met Phe Val Tyr Ser Leu Glu Glu Arg Val Glu Asn Asn 65 70 75 80 Ser Val Pro Ser Arg Phe Ser Pro Glu Cys Pro Asn Ser Ser His Leu 85 90 95 Phe Leu His Leu His Thr Leu Gln Pro Glu Asp Ser Ala Leu Tyr Leu 100 105 110 Cys Ala Ser Ser Gln Asp Arg Pro Asn Tyr Tyr Gly Tyr Thr Phe Gly 115 120 125 Ser Gly Thr Arg Leu Thr Val Val 130 135 <210> 109 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2705 - codon optimized <400> 109 atgatgaagt ccctgagagt gctgctggtc atcctgtggc tgcagctgtc ttgggtctgg 60 tcccagcaga aagaagtgga acaggaccct ggacctctga gcgttccaga aggcgccatc 120 gtcagcctga attgcaccta cagcaacagc gccttccagt acttcatgtg gtacagacag 180 tactcccgga agggccccga gctgctgatg tacacataca gcagcggcaa caaagaggac 240 ggccggttta cagcccaggt ggacaagagc agcaagtaca tctccctgtt catccgggac 300 agccagccta gcgatagcgc cacatacctg tgcgccatgt ctggcacagg cggcttcaag 360 accatcttcg gagccggcac acggctgttc gtgaaggcca ac 402 <210> 110 <211> 408 <212> DNA <213> Artificial sequence <220> <223> Variable region of beta chain of TCR2705 - codon optimized <400> 110 atgggatgta gactgctgtg ttgtgccgtg ctgtgtctgc ttggagctgg cgaactggtg 60 cctatggaaa ccggcgtgac ccagacacct agacacctgg tcatgggcat gacaaacaag 120 aaaagcctga agtgcgagca gcacctgggc cacaatgcca tgtactggta caagcagagc 180 gccaagaaac ccctggaact gatgttcgtg tacagcctgg aagagagggt cgagaacaac 240 agcgtgccca gcagattcag ccctgagtgc cctaatagca gccacctgtt tctgcatctg 300 cacaccctgc agcctgagga ctctgccctg tatctgtgtg ccagcagcca ggacagaccc 360 aactactacg gctacacctt tggcagcggc accagactga ccgtggtg 408 <210> 111 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2709 <400> 111 Thr Ile Ser Gly Thr Asp Tyr 1 5 <210> 112 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2709 <400> 112 Gly Leu Thr Ser Asn 1 5 <210> 113 <211> 16 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2709 <400> 113 Cys Ile Leu Arg Asp Arg Tyr Gly Gly Ser Gln Gly Asn Leu Ile Phe 1 5 10 15 <210> 114 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2709 <400> 114 Met Asn His Glu Tyr 1 5 <210> 115 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2709 <400> 115 Ser Val Gly Glu Gly Thr 1 5 <210> 116 <211> 18 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2709 <400> 116 Cys Ala Ser Ser Tyr Trp Pro Thr Thr Gly Glu Ser Thr Asp Thr Gln 1 5 10 15 Tyr Phe <210> 117 <211> 131 <212> PRT <213> Homo sapiens <220> <223> Variable region of the alpha chain of TCR2709 <400> 117 Met Lys Leu Val Thr Ser Ile Thr Val Leu Leu Ser Leu Gly Ile Met 1 5 10 15 Gly Asp Ala Lys Thr Thr Gln Pro Asn Ser Met Glu Ser Asn Glu Glu 20 25 30 Glu Pro Val His Leu Pro Cys Asn His Ser Thr Ile Ser Gly Thr Asp 35 40 45 Tyr Ile His Trp Tyr Arg Gln Leu Pro Ser Gln Gly Pro Glu Tyr Val 50 55 60 Ile His Gly Leu Thr Ser Asn Val Asn Asn Arg Met Ala Ser Leu Ala 65 70 75 80 Ile Ala Glu Asp Arg Lys Ser Ser Thr Leu Ile Leu His Arg Ala Thr 85 90 95 Leu Arg Asp Ala Ala Val Tyr Tyr Cys Ile Leu Arg Asp Arg Tyr Gly 100 105 110 Gly Ser Gln Gly Asn Leu Ile Phe Gly Lys Gly Thr Lys Leu Ser Val 115 120 125 Lys Pro Asn 130 <210> 118 <211> 136 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2709 <400> 118 Met Ser Leu Gly Leu Leu Cys Cys Gly Ala Phe Ser Leu Leu Trp Ala 1 5 10 15 Gly Pro Val Asn Ala Gly Val Thr Gln Thr Pro Lys Phe Arg Val Leu 20 25 30 Lys Thr Gly Gln Ser Met Thr Leu Leu Cys Ala Gln Asp Met Asn His 35 40 45 Glu Tyr Met Tyr Trp Tyr Arg Gln Asp Pro Gly Met Gly Leu Arg Leu 50 55 60 Ile His Tyr Ser Val Gly Glu Gly Thr Thr Ala Lys Gly Glu Val Pro 65 70 75 80 Asp Gly Tyr Asn Val Ser Arg Leu Lys Lys Gln Asn Phe Leu Leu Gly 85 90 95 Leu Glu Ser Ala Ala Pro Ser Gln Thr Ser Val Tyr Phe Cys Ala Ser 100 105 110 Ser Tyr Trp Pro Thr Thr Gly Glu Ser Thr Asp Thr Gln Tyr Phe Gly 115 120 125 Pro Gly Thr Arg Leu Thr Val Leu 130 135 <210> 119 <211> 393 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2709 - codon optimized <400> 119 atgaagctgg tcaccagcat cacagtgctg ctgagcctgg gaattatggg cgacgccaag 60 accacacagc ccaacagcat ggaaagcaac gaagaggaac ccgtgcatct gccctgcaac 120 cacagcacaa tcagcggcac cgactacatc cactggtata gacagctgcc ctctcagggc 180 cccgagtatg tgattcacgg actgaccagc aacgtgaaca accggatggc ctctctggcc 240 attgccgagg acagaaagag cagcaccctg atcctgcaca gagccacact gagagatgcc 300 gccgtgtact actgcatcct gcgggataga tacggcggca gccagggcaa tctgatcttt 360 ggcaagggca ccaagctgag cgtgaagccc aac 393 <210> 120 <211> 408 <212> DNA <213> Artificial Sequence <220> <223> Variable region of beta chain of TCR2709 - codon optimized <400> 120 atgtctcttg gattgctttg ctgcggcgcc ttcagcctgc tttgggctgg acctgttaat 60 gccggcgtga cccagacacc taagttccgg gtgctgaaaa ccggccagag catgacactg 120 ctgtgcgccc aggacatgaa ccacgagtac atgtattggt acagacagga ccccggcatg 180 ggcctgagac tgatccacta ttctgtcggc gagggcacca cagccaaagg cgaagttcct 240 gacggctaca acgtgtcccg gctgaagaag cagaacttcc tgctgggcct cgagtctgcc 300 gctccatctc agaccagcgt gtacttctgt gccagcagct actggcctac caccggcgag 360 tctaccgaca cacagtattt cggccctggc accagactga ccgtgctg 408 <210> 121 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2716 <400> 121 Thr Ser Glu Asn Asn Tyr Tyr 1 5 <210> 122 <211> 8 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2716 <400> 122 Gln Glu Ala Tyr Lys Gln Gln Asn 1 5 <210> 123 <211> 17 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2716 <400> 123 Cys Ala Phe Met Lys Pro Tyr Ser Gly Gly Gly Ala Asp Gly Leu Thr 1 5 10 15 Phe <210> 124 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2716 <400> 124 Pro Arg His Asp Thr 1 5 <210> 125 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2716 <400> 125 Phe Tyr Glu Lys Met Gln 1 5 <210> 126 <211> 16 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2716 <400> 126 Cys Ala Ser Ser Leu Ala Gly Thr Thr Val Tyr Asn Glu Gln Phe Phe 1 5 10 15 <210> 127 <211> 139 <212> PRT <213> Homo sapiens <220> <223> Variable region of alpha chain of TCR2716 <400> 127 Met Thr Arg Val Ser Leu Leu Trp Ala Val Val Val Ser Thr Cys Leu 1 5 10 15 Glu Ser Gly Met Ala Gln Thr Val Thr Gln Ser Gln Pro Glu Met Ser 20 25 30 Val Gln Glu Ala Glu Thr Val Thr Leu Ser Cys Thr Tyr Asp Thr Ser 35 40 45 Glu Asn Asn Tyr Tyr Leu Phe Trp Tyr Lys Gln Pro Pro Ser Arg Gln 50 55 60 Met Ile Leu Val Ile Arg Gln Glu Ala Tyr Lys Gln Gln Asn Ala Thr 65 70 75 80 Glu Asn Arg Phe Ser Val Asn Phe Gln Lys Ala Ala Lys Ser Phe Ser 85 90 95 Leu Lys Ile Ser Asp Ser Gln Leu Gly Asp Thr Ala Met Tyr Phe Cys 100 105 110 Ala Phe Met Lys Pro Tyr Ser Gly Gly Gly Ala Asp Gly Leu Thr Phe 115 120 125 Gly Lys Gly Thr His Leu Ile Ile Gln Pro Tyr 130 135 <210> 128 <211> 144 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2716 <400> 128 Met Leu Ser Pro Asp Leu Pro Asp Ser Ala Trp Asn Thr Arg Leu Leu 1 5 10 15 Cys His Val Met Leu Cys Leu Leu Gly Ala Val Ser Val Ala Ala Gly 20 25 30 Val Ile Gln Ser Pro Arg His Leu Ile Lys Glu Lys Arg Glu Thr Ala 35 40 45 Thr Leu Lys Cys Tyr Pro Ile Pro Arg His Asp Thr Val Tyr Trp Tyr 50 55 60 Gln Gln Gly Pro Gly Gln Asp Pro Gln Phe Leu Ile Ser Phe Tyr Glu 65 70 75 80 Lys Met Gln Ser Asp Lys Gly Ser Ile Pro Asp Arg Phe Ser Ala Gln 85 90 95 Gln Phe Ser Asp Tyr His Ser Glu Leu Asn Met Ser Ser Leu Glu Leu 100 105 110 Gly Asp Ser Ala Leu Tyr Phe Cys Ala Ser Ser Leu Ala Gly Thr Thr 115 120 125 Val Tyr Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 130 135 140 <210> 129 <211> 417 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2716 - codon optimized <400> 129 atgaccagag tgtctctgct gtgggccgtc gtggtgtcca catgtctgga atctggcatg 60 gcccagacag tgacccagag ccagcctgag atgtctgtgc aagaggccga gactgtgacc 120 ctgtcctgca cctacgatac cagcgagaac aactactacc tgttctggta caagcagcct 180 cctagccggc agatgatcct ggtcatcaga caagaggcct ataagcagca gaacgccacc 240 gagaacagat tcagcgtgaa cttccagaag gccgccaaga gcttcagcct gaagatcagc 300 gatagccagc tgggagacac cgccatgtat ttctgcgcct ttatgaagcc ctacagcggc 360 ggaggtgccg atggcctgac atttggaaag ggcacccacc tgattatcca gccgtac 417 <210> 130 <211> 432 <212> DNA <213> Artificial sequence <220> <223> Variable region of beta chain of TCR2716 - codon optimized <400> 130 atgctttctc cagatctgcc tgacagcgcc tggaacacca gactgctgtg tcacgtgatg 60 ctgtgtctgc tgggagccgt gtctgttgcc gctggcgtta tccagtctcc tcggcacctg 120 atcaaagaga agagagagac agccacactg aagtgctacc ccattccacg gcacgacacc 180 gtgtactggt atcagcaagg cccaggccag gatcctcagt tcctgatcag cttctacgag 240 aagatgcaga gcgacaaggg cagcatcccc gacagatttt ctgcccagca gttcagcgac 300 taccacagcg agctgaacat gagcagcctg gaactgggcg atagcgccct gtacttttgt 360 gcctcttctc tggccggcac cacagtgtac aacgagcagt ttttcggccc tggcaccagg 420 ctgaccgtgc tg 432 <210> 131 <211> 7 <212> PRT <213> Homo sapiens <220> <223> CDR1 alpha chain TCR2719 <400> 131 Asn Ile Ala Thr Asn Asp Tyr 1 5 <210> 132 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR2 alpha chain TCR2719 <400> 132 Gly Tyr Lys Thr Lys 1 5 <210> 133 <211> 13 <212> PRT <213> Homo sapiens <220> <223> CDR3 alpha chain TCR2719 <400> 133 Cys Leu Val Gly Ala Asp Ser Asn Tyr Gln Leu Ile Trp 1 5 10 <210> 134 <211> 5 <212> PRT <213> Homo sapiens <220> <223> CDR1 beta chain TCR2719 <400> 134 Ser Gly Asp Leu Ser 1 5 <210> 135 <211> 6 <212> PRT <213> Homo sapiens <220> <223> CDR2 beta chain TCR2719 <400> 135 Tyr Tyr Asn Gly Glu Glu 1 5 <210> 136 <211> 14 <212> PRT <213> Homo sapiens <220> <223> CDR3 beta chain TCR2719 <400> 136 Cys Ala Ser Ser Pro Gly Gly Gly Ala Tyr Glu Gln Tyr Phe 1 5 10 <210> 137 <211> 128 <212> PRT <213> Homo sapiens <220> <223> Variable region of the alpha chain of TCR2719 <400> 137 Met Arg Gln Val Ala Arg Val Ile Val Phe Leu Thr Leu Ser Thr Leu 1 5 10 15 Ser Leu Ala Lys Thr Thr Gln Pro Ile Ser Met Asp Ser Tyr Glu Gly 20 25 30 Gln Glu Val Asn Ile Thr Cys Ser His Asn Asn Ile Ala Thr Asn Asp 35 40 45 Tyr Ile Thr Trp Tyr Gln Gln Phe Pro Ser Gln Gly Pro Arg Phe Ile 50 55 60 Ile Gln Gly Tyr Lys Thr Lys Val Thr Asn Glu Val Ala Ser Leu Phe 65 70 75 80 Ile Pro Ala Asp Arg Lys Ser Ser Thr Leu Ser Leu Pro Arg Val Ser 85 90 95 Leu Ser Asp Thr Ala Val Tyr Tyr Cys Leu Val Gly Ala Asp Ser Asn 100 105 110 Tyr Gln Leu Ile Trp Gly Ala Gly Thr Lys Leu Ile Ile Lys Pro Asp 115 120 125 <210> 138 <211> 132 <212> PRT <213> Homo sapiens <220> <223> Variable region of the beta chain of TCR2719 <400> 138 Met Gly Phe Arg Leu Leu Cys Cys Val Ala Phe Cys Leu Leu Gly Ala 1 5 10 15 Gly Pro Val Asp Ser Gly Val Thr Gln Thr Pro Lys His Leu Ile Thr 20 25 30 Ala Thr Gly Gln Arg Val Thr Leu Arg Cys Ser Pro Arg Ser Gly Asp 35 40 45 Leu Ser Val Tyr Trp Tyr Gln Gln Ser Leu Asp Gln Gly Leu Gln Phe 50 55 60 Leu Ile Gln Tyr Tyr Asn Gly Glu Glu Arg Ala Lys Gly Asn Ile Leu 65 70 75 80 Glu Arg Phe Ser Ala Gln Gln Phe Pro Asp Leu His Ser Glu Leu Asn 85 90 95 Leu Ser Ser Leu Glu Leu Gly Asp Ser Ala Leu Tyr Phe Cys Ala Ser 100 105 110 Ser Pro Gly Gly Gly Ala Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg 115 120 125 Leu Thr Val Thr 130 <210> 139 <211> 384 <212> DNA <213> Artificial Sequence <220> <223> Variable region of alpha chain of TCR2719 - codon optimized <400> 139 atgagacagg tggccagagt gatcgtgttc ctgacactga gcaccctgag cctggccaag 60 accacacagc ccatcagcat ggacagctac gagggccaag aagtgaacat cacctgtagc 120 cacaacaata tcgccaccaa cgactacatc acgtggtatc aacagttccc cagtcaaggc 180 cctcggttca tcatccaagg ctacaagacc aaagtgacca acgaggtggc ctctctgttc 240 atccccgccg acagaaagag cagcaccctg tctctgccta gagtgtccct gagcgatacc 300 gccgtgtact actgtctcgt gggcgccgac tctaactacc agctgatttg gggagccggc 360 accaagctga tcatcaagcc cgac 384 <210> 140 <211> 396 <212> DNA <213> Artificial sequence <220> <223> Variable region of beta chain of TCR2719 - codon optimized <400> 140 atgggtttta gactgctgtg ctgcgtggcc ttctgtctgc ttggagctgg ccctgtggat 60 agcggcgtta cccagacacc taagcacctg atcacagcca caggccagcg cgtgaccctg 120 agatgttctc ctagaagcgg cgacctgagc gtgtactggt atcagcagtc tctggaccag 180 ggcctgcagt tcctgatcca gtactacaac ggcgaggaaa gagccaaggg caacatcctg 240 gaacggttca gcgcccagca gttcccagat ctgcacagcg agctgaacct gagcagcctg 300 gaactgggag atagcgccct gtacttctgc gccagttctc ctggtggcgg agcctacgag 360 cagtattttg gccctggcac cagactgacc gtgacc 396 <210> 141 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of CMKLR1 <400> 141 Leu Pro Ile His Ile Thr Tyr Ala Ala Met 1 5 10 <210> 142 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of SLCO2B1 <400> 142 Arg Pro Arg Met Ile Gly Tyr Gly Ala Ile 1 5 10 <210> 143 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of RP11-1220K2.2 <400> 143 Arg Pro Ala Met Ile Pro Tyr Trp Ala Leu 1 5 10 <210> 144 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of CHRNB1 <400> 144 Val Pro Ile Ile Ile Lys Tyr Leu Met Phe 1 5 10 <210> 145 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of PTPRG <400> 145 Arg Pro Val Pro Ile Ser Tyr His Gln Leu 1 5 10 <210> 146 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of PGBD1 <400> 146 Lys Pro Ile Arg Ile Gly Tyr Lys Ile Trp 1 5 10 <210> 147 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of EHBP1 <400> 147 Arg Pro Asp Leu Ile Asp Tyr Lys Ser Leu 1 5 10 <210> 148 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of GPR1 <400> 148 Leu Pro Leu Tyr Ile Ser Tyr Val Ala Met 1 5 10 <210> 149 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of GPR183 <400> 149 Leu Pro Thr Arg Ile Ala Tyr Tyr Ala Met 1 5 10 <210> 150 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of TMEM230 <400> 150 Thr Pro Pro Lys Ile Pro Tyr Lys Ala Ile 1 5 10 <210> 151 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of EFEMP1 <400> 151 Leu Pro Gln Ser Ile Val Tyr Lys Tyr Met 1 5 10 <210> 152 <211> 10 <212> PRT <213> Homo sapiens <220> <223> Epitope of ADAMTSL2 <400> 152 Arg Pro Gln Pro Ile Tyr Tyr Gly Phe Ser 1 5 10 BRIEF DESCRIPTION OF THE DRAWINGS

[0113] Figure 1 : Generation of Mutation-Specific T Cells

[0114] A. Schematic representation of the methodology for generating mutation-specific T cells. B. Representative streptamer staining of clone-10 (TCR1610) after a single restimulation. C. Selective reactivity of a large number of T cell clones to mutant peptides was detected by co-culturing overnight with autologous PBMCs loaded with peptides prior to FACS isolation of streptamer-positive cells. Reactivity was measured by IFNγ ELISA.

[0115] Figure 2 : Identification of Mutation-Specific T Cell Receptors (TCRs)

[0116] Representative construct of the TCR gene cassette.

[0117] Figure 3 : TCR Affinity Analysis

[0118] A. / C. Non-linear curve analysis of the IFNγ response of TCR-transduced CD8+ T cells from healthy donors when co-cultured with K562 cells transduced with HLA*B07:02 and loaded with different concentrations of mutant peptide (SEQ ID NO:2). The detectable concentration of the response to the mutant peptide can be as low as 10 -4 μg / ml, with K D values in the nanomolar (high affinity) range. B. / D. IFNγ response to the corresponding wild-type (WT) peptide (ID NO:6). The mutant-specific TCR showed more than 10,000-fold higher affinity for the mutant peptide.

[0119] Figure 4: Mutant-Specific Activation of TCR-Engineered T Cells

[0120] A. Mutant-specific activation of T cells transduced with TCR2207 against K562 cells in the presence or absence of HLA-B7 expression, also transduced with virus to express full-length wild-type or mutant (L265P) MYD88, analyzed by flow cytometry after 16 hours of co-culture. B. / C. Comparative mutant-specific activation analysis of TCR-transduced T cells. The IFNγ response measured by ELISA was a mutant-specific and HLA-B7-restricted response. The epitope can be processed and presented by human cells.

[0121] Figure 5: Mutant-Specific Cytotoxicity of TCR-Engineered T Cells

[0122] A. The viability of HLA-B7-positive target cells co-cultured with T cells expressing the TCR of the present invention for 16 hours was analyzed by flow cytometry (shown: 3 highest affinity TCRs). Cells were gated as GFP-positive as a reporter gene for wild-type or mutant MYD88 expression, and viability was analyzed by combining intracellular staining of activated Caspase-3 (a-Caspase-3) with fixable dead cell staining. The lower left quadrant provides the number of viable cells. B. / C. Viability of target cells in comparative cytotoxicity analysis of different TCR-transduced T cells. Target cells expressing the mutant and HLA*B07:02 were specifically killed by TCR-transduced T cells. The cytotoxicity intensity was closely related to the TCR affinity.

[0123] Figure 6 : Mutant-Specific Activation of TCR-Engineered T Cells Against Lymphoma Cell Lines

[0124] A. Flow cytometric activation analysis of T cells transduced with one of the two highest affinity TCRs after 16 h of co - culture with OCI - Ly3 (ABC - like DLBCL, homozygous MYD88 - L265P) and HBL - 1 (ABC - like DLBCL, heterozygous MYD88 - L265P) lymphoma cell lines. Since both cell lines are HLA - B7 negative, they were virally transduced to express the antigen (shown as: "Cell line_B7"). OCI - Ly3 cells transduced with HLA - B7 were highly recognized by TCR - engineered T cells. A weaker response was observed against heterozygous mutant HBL - 1 cells, which was slightly improved when target cells were pre - treated with 50 ng / ml human IFNγ overnight before co - culture, which is known to improve proteasomal processing of peptides and MHC presentation. B. Mutation - specific and HLA - B7 - restricted activation of TCR2304 - transduced T cells.

[0125] Figure 7 : Mutation - specific cytotoxicity against lymphoma cell lines

[0126] A. Flow cytometric viability analysis of OCI - Ly3 lymphoma cells after 16 h of co - culture with TCR2304 - transduced T cells (shown in Figure 5). B. Mutation - specific killing of TCR2304 - transduced T cells. C. Antigen - induced proliferation of TCR2304 - transduced T cells after 72 h of co - culture with HLA - B7 - positive OCI - Ly3 cells. T cells were labeled with CSFE before co - culture to track proliferation.

[0127] Figure 8 : Characterization of peptide - MHC binding behavior of TCRs by alanine scanning

[0128] A. Alanine scanning was performed by substituting each amino acid in the mutant epitope (SEQ ID NO:2) with alanine one by one to study the effect of individual amino acids on the peptide - MHC - TCR relationship. All peptides were separately loaded onto K562 cells expressing HLA - B7 and co - cultured with TCR - transduced T cells for 16 h to measure IFNγ production by ELISA. B. Amino acid positions that affect the IFNγ response by more than 50% were considered important for the peptide - MHC - TCR relationship, and this binding motif was used for off - target cross - reactivity prediction using the online tool Expitope (Jaravine et al., 2017). Peptides (SEQ ID NO:141 - 152) predicted to bind HLA - B7 by this analysis for TCR2304 were re - loaded onto K562 cells expressing HLA - B7 and co - cultured with TCR - transduced T cells from 3 different donors for 16 h. No cross - reactivity against these peptides was observed.

[0129] Figure 9 :

[0130] A. After co - culturing with SU - DHL - 6 (GBC - like DLBCL, wild - type MYD88), OCI - Ly3 (ABC - like DLBCL, homozygous MYD88 - L265P), and TMD8 (ABC - like DLBCL, heterozygous MYD88 L265P) lymphoma cell lines for 16 hours, the activation of TCR2304 - transduced T cells was analyzed by IFNγ ELISA. Since all cell lines were negative for HLA - B7, they could be virally transduced to express this antigen (shown as: "Cell line_B7"). OCI - Ly3 and TMD8 cells transduced with HLA - B7 were highly recognized by TCR - engineered T cells. B. After co - culturing with TCR2304 - transduced T cells for 16 hours, flow cytometry viability analysis of lymphoma cells (as shown in Figure 5) showed mutant - specific killing. Example

[0131] Example 1: Generation of mutant - specific T cells

[0132] PBMCs were isolated from the blood of healthy HLA - B7 - positive donors. Monocytes were separated by plastic adhesion to generate dendritic cells (DCs), which were then cultured in RPMI with 1% human serum containing 800 IU / ml GM - CSF and 10 ng / ml IL - 4 for 3 days. Subsequently, immature dendritic cells (imDCs) were cultured overnight in the presence of 10 ng / ml LPS and 50 ng / ml interferon - γ (IFNγ) to mature them. Then, mature dendritic cells (mDCs) were loaded with the mutant peptide (RPIPIKYKAM, SEQ ID NO: 2) and used to prime autologous CD8 - positive naive T cells (5×10 5 T cells / well, with donor - dependent DC - T cell ratio variation). After 10 days, cells in each well were stained with a specific streptamer (HLA*B07:02 - RPIPIKYKAM), or T cell activation markers (such as CD137 (4 - 1BB)) were stained after short (∼6 - hour) peptide restimulation. Positive - stained wells were restimulated with peptide - loaded autologous PBMCs for expansion. In this case, it was necessary to obtain sufficient cells for FACS isolation.

[0133] A schematic of the methodology for generating mutant - specific T cells is shown as Figure 1 A. Figure 1 B shows the representative streptamer staining of clone - 10 (TCR1610) after a single restimulation.

[0134] Prior to FACS isolation of streptamer - positive or peptide - reactive cells, a large number of T - cell clones with selective reactivity to mutant peptides were detected by overnight co - culture with peptide - loaded autologous PBMCs. Responses were measured by IFNγ ELISA ( Figure 1 C).

[0135] Example 2: Identification of mutant - specific T - cell receptors (TCRs)

[0136] After the final IFNγ ELISA assay, viable CD8 and streptamer - positive cells were separately isolated from each reactive T - cell clone by FACS. Total RNA isolation was performed. TCRα and β genes were amplified by 5′ - RACE PCR and cloned. Multiple bacterial clones from each TCR chain were sequenced to analyze T - cell clonality. Table 1 shows the CDR3s, their sequence numbers, and gene subtypes of MyD88 - L265P mutant - specific TCRs, and Table 2 shows a list of the amino - acid sequences of CDR1 and CDR2.

[0137] Table 1

[0138]

[0139]

[0140]

[0141] Table 2

[0142]

[0143]

[0144] The sequences of the identified variable domains were combined with those of murine constant domains for experimental characterization of the TCRs and synthesized by codon optimization for expression in human cells. TCR gene kits encoding TRBV binding to murine TRBC and TRAV binding to TRAC separated by the p2A signal were constructed as described in detail by Obenaus et al. (2015) and Sommermeyer et al. (2010) (see Figure 2 ).

[0145] Example 3: TCR affinity analysis

[0146] Peripheral CD8+ T cells from HLA-B7 positive healthy donors were successfully transduced to express mutant-specific TCRs, showed no signs of fratricide, and were co-cultured with K562 cells transduced with HLA*B07:02 and loaded with different concentrations of mutant peptides. IFNγ responses were measured by ELISA. Figure 3 A / C shows non-linear curve analysis of the IFNγ response of TCR-engineered T cells to mutant peptide titration. The detectable concentration of the response to the mutant peptide was as low as 10 -4 μg / ml, and the K D value was in the nanomolar (high affinity) range. Figure 3 B / D shows non-linear curve analysis of the IFNγ response to titration of the corresponding wild-type peptide. The mutant-specific TCR showed more than 10,000-fold affinity for the mutant peptide. Table 3 shows the affinities of the different TCRs analyzed.

[0147] Table 3: Affinity of TCRs for SEQ ID NO:2 in the context of HLA-B*07:02 (shown as K D ). The molecular weight of peptide SEQ IDNO:2 is 1216.54 g / mol.

[0148] TCR <![CDATA[K D (μg / ml)]]> <![CDATA[K D (M)*]]> 2207 0.003 <![CDATA[2.4×10 -9 > 2304 0.003 <![CDATA[2.4×10 -9 > 2205 0.004 <![CDATA[3.2×10 -9 <!-- 17 -->]]> 1605 0.009 <![CDATA[7.4×10 -9 > 1610 0.009 <![CDATA[7.4×10 -9 > 2202 0.033 <![CDATA[2.7×10 -8 > 2219 0.123 <![CDATA[1×10 -7 > 1336 0.387 <![CDATA[3.1×10 -7 > 2211 0.560 <![CDATA[4.6×10 -7 > 2705 0.020 <![CDATA[1.6×10 -8 > 2709 0.102 <![CDATA[8.3×10 -8 > 2716 0.099 <![CDATA[8.1×10 -8 > 2719 0.024 <![CDATA[1.9×10 -8 >

[0149] TCR2304 and TCR2207 had the highest affinity for the mutant peptide, with a K D of 0.003 g / ml for SEQ ID NO:2, which is equivalent to 2.4 nM.

[0150] Example 4: Mutant-specific activation of TCR-engineered T cells

[0151] K562 cells with or without HLA-B7 were virally transduced to express full-length wild-type or mutant (L265P) MYD88, which was coupled by p2A to express the marker GFP and used as an artificial target cell to evaluate the cytotoxic reactivity of TCR-engineered T cells. When co-cultured for 16 hours, 6 TCRs achieved recognition of target cells expressing mutant MyD88 without prior peptide loading, indicating that the epitope could be successfully processed and presented by human cells.

[0152] Figure 4A shows mutant-specific activation of T cells transduced with TCR2207, one of the TCRs, by flow cytometry analysis of the activation marker CD137. Figures 4B / C show comparative mutant-specific IFNγ responses of T cells transduced with different TCRs measured by ELISA, showing mutant-specific and HLA-B7-restricted responses.

[0153] Example 5: Mutation-specific cytotoxicity of TCR-transduced T cells

[0154] TCR-transduced T cells were co-cultured with K562 cells expressing full-length wild-type or mutant MYD88 linked to GFP (as an expression marker) with p2A for 16 hours under the control of the same promoter with or without HLA-B7. Target cells expressing the mutation and HLA*B07:02 were specifically killed by TCR-transduced T cells. Figure 5A shows the survival rate of HLA-B7-positive target cells co-cultured with T cells expressing one of the three highest-affinity TCRs for 16 hours, which was analyzed by flow cytometry. Cells were gated as GFP-positive as a reporter for wild-type or mutant MYD88 expression, and viability was analyzed by combining intracellular staining of activated Caspase-3 (a-Caspase-3) with fixable dead cell staining. Figures 5B / C show the viability of target cells in a comparative cytotoxicity analysis of T cells transduced with different TCRs.

[0155] Example 6: Mutation-specific activation of TCR-engineered T cells against lymphoma cell lines

[0156] To study the anti-mutant functional potential of TCR-engineered T cells at more physiological expression levels, the activation of T cells transduced with one of the two highest-affinity TCRs was analyzed by flow cytometry after co-culturing with OCI-Ly3 (ABC-like DLBCL, homozygous MYD88-L265P) or HBL-1 (ABC-like DLBCL, heterozygous MYD88-L265P) lymphoma cell lines for 16 hours. Since both cell lines are negative for HLA-B7, they were virally transduced to express this antigen (shown as: "cell line_B7"). OCI-Ly3 cells transduced with HLA-B7 were highly recognized by TCR-engineered T cells. A weaker response was observed against heterozygous mutant HBL-1 cells, which was slightly improved when target cells were pre-treated with 50 ng / ml human IFNγ overnight before co-culture. IFNγ is known to improve proteasomal processing of peptides and MHC presentation in some cases.

[0157] Figure 6 A shows flow cytometry analysis of T cell responses against OCI-Ly3 and HBL-1 cells. Figure 6 B shows mutation-specific and HLA-B7-restricted activation of T cells transduced with TCR2304 against OCI-Ly3 cells.

[0158] Example 7: Mutation-specific cytotoxicity against lymphoma cell lines

[0159] T cells transduced with TCR2304 were labeled with CSFE and co-cultured with OCI-Ly3 cells with or without HLA-B7. The viability of the target lymphoma cells was analyzed as described in Example 5 above.

[0160] Figure 7 A shows the survival rate of OCI-Ly3 cells with or without HLA-B7 expression after co-culture with TCR2304-transduced T cells for 16 hours. Figure 7 B shows the killing effect of TCR2304-transduced T cells on mutant-specific lymphoma cells. Figure 7 C shows the antigen-induced proliferation of TCR2304-transduced T cells after co-culture with HLA-B7-positive OCI-Ly3 cells for 72 hours, as a decrease in CSFE fluorescence intensity indicates that cell division occurs only in TCR-transduced cells.

[0161] Example 8: Characterization of Peptide-MHC Binding Behavior of TCRs by Alanine Scanning

[0162] Alanine scanning was performed by substituting each amino acid in the mutant epitope (SEQ ID NO: 2) with alanine one by one to study the effect of individual amino acids on the peptide-MHC-TCR relationship. All peptides were separately loaded onto K562 cells expressing HLA-B7 and co-cultured with TCR-transduced T cells for 16 hours. Different numbers and groups of amino acids were observed and considered essential for the recognition of different TCRs (binding motifs). However, proline at position 2, which reflects the amino acid substitution L265P on mutant MyD88, was absolutely essential for all TCRs, demonstrating the specificity of TCRs for the mutation ( Figure 8 A). As part of the safety screening, the possibility of cross-reactivity caused by binding sequence similarity to other human proteins was analyzed separately using the online tool Expitope (Jaravine et al., 2017). For TCR2304, this analysis showed that there were 12 peptides in the human proteome with binding motif similarity (up to 5 mismatched positions) and different predicted affinities for HLA-B7 (SEQ ID NOs: 141-152). Again, all these peptides were loaded onto K562 cells expressing HLA-B7 for co-culture with TCR-transduced T cells from 3 different donors. No TCR recognition against any peptide was observed ( Figure 8 B).

[0163] Example 9:

[0164] To better understand TCR recognition and T cell function against cells naturally carrying the MYD88 L265P mutation, TCR2304-transduced T cells from 3 different healthy donors were co-cultured with DLBCL cell lines for 16 hours; SU-DHL-6 (wild-type MYD88), OCI-Ly3 (homozygous MYD88 L265P), or TMD8 (heterozygous MYD88 L265P), with or without HLA-B7 expression.

[0165] Figure 9 A shows mutant-specific recognition of OCI-Ly3 and TMD8 cell lines with HLA-B7 expression. The SU-DHL-6 control cell line with HLA-B7 was recognized only when loaded with mutant peptides prior to co-culture.

[0166] Figure 9 B shows effective mutant-specific and HLA-restricted killing of OCI-Ly3 and TMD8 cell lines by TCR-transduced T cells.

Claims

1. A nucleic acid, wherein the nucleic acid encodes at least one TCRα-chain and β-chain construct of a TCR construct that is capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02, a) wherein the TCRα-chain construct comprises the CDR1 sequence of SEQ ID NO:91, the CDR2 sequence of SEQ ID NO:92, and the CDR3 sequence of SEQ ID NO:93, and the TCRβ-chain construct comprises the CDR1 sequence of SEQ ID NO:94, the CDR2 sequence of SEQ ID NO:95, and the CDR3 sequence of SEQ ID NO:96; or b) wherein the TCRα-chain construct comprises the CDR1 sequence of SEQ ID NO:11, the CDR2 sequence of SEQ ID NO:12, and the CDR3 sequence of SEQ ID NO:13, and wherein the TCRβ-chain construct comprises the CDR1 sequence of SEQ ID NO:14, the CDR2 sequence of SEQ ID NO:15, and the CDR3 sequence of SEQ ID NO:

16.

2. The nucleic acid according to claim 1, wherein the TCRα-chain construct comprises the CDR1 sequence of SEQ ID NO:91, the CDR2 sequence of SEQ ID NO:92, and the CDR3 sequence of SEQ ID NO:93, and wherein the TCRβ-chain construct comprises the CDR1 sequence of SEQ ID NO:94, the CDR2 sequence of SEQ ID NO:95, and the CDR3 sequence of SEQ ID NO:

96.

3. The nucleic acid according to claim 2, wherein, The TCRα-chain construct comprises a variable region having at least 90% sequence identity with SEQ ID NO:97, and / or wherein the TCRβ-chain construct comprises a variable region having at least 90% sequence identity with SEQ ID NO:

98.

4. The nucleic acid according to claim 1, wherein the TCRα-chain construct comprises the CDR1 sequence of SEQ ID NO:11, the CDR2 sequence of SEQ ID NO:12, and the CDR3 sequence of SEQ ID NO:13, and wherein the TCRβ-chain construct comprises the CDR1 sequence of SEQ ID NO:14, the CDR2 sequence of SEQ ID NO:15, and the CDR3 sequence of SEQ ID NO:

16.

5. The nucleic acid according to claim 4, wherein, The TCRα-chain construct comprises a variable region having at least 90% sequence identity with SEQ ID NO:17, and / or wherein the TCRβ-chain construct comprises a variable region having at least 90% sequence identity with SEQ ID NO:

18.

6. The nucleic acid according to claim 1, wherein the TCRα-chain construct and / or the TCRβ-chain construct further comprises a constant region selected from a human constant region, a murine constant region, or a chimeric constant region.

7. The nucleic acid according to any one of claims 1-6, wherein the nucleic acid encodes at least one TCRα chain and TCRβ chain construct of the TCR construct, and the nucleic acid is selected from viral vectors, transposons or vectors suitable for CRISPR / CAS-based recombination.

8. A protein encoded by the nucleic acid according to any one of claims 1-7.

9. A host cell comprising the nucleic acid according to any one of claims 1-7 and / or the protein according to claim 8.

10. The host cell according to claim 9, wherein the host cell is a human CD8+ T cell.

11. A pharmaceutical composition comprising: a) the nucleic acid according to any one of claims 1-7, wherein the nucleic acid encodes a TCR construct capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02; or b) the protein according to claim 8, wherein the protein comprises a TCR construct capable of specifically binding to the MYD88 L265P peptide of SEQ ID NO:2 in the presence of HLA-B*07:02; or c) the host cell according to claim 10, wherein the host cell expresses a TCR construct capable of specifically binding to the MYD88 L265P peptide containing SEQ ID NO:2 in the presence of HLA-B*07:

02.

12. Use of the pharmaceutical composition according to claim 11 in the preparation of a medicament for diagnosing or treating a patient suspected of containing cells expressing the MYD88 L265P protein, wherein the patient has non-Hodgkin B-cell lymphoma, and the non-Hodgkin B-cell lymphoma is activated B-cell type diffuse large B-cell lymphoma (ABC-DLBCL).

13. The use according to claim 12, wherein the medicament is for immunotherapy, and the immunotherapy is selected from adoptive T-cell therapy or TCR gene therapy for patients containing cells expressing the MYD88 L265P protein.

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