阅读说明：本技术 Ebv表位高亲和力t细胞受体 (EBV epitope high affinity T cell receptors ) 是由 王飞 赵正琦 王石雷 尹悦露 李波 侯勇 于 2018-12-27 设计创作，主要内容包括：本发明公开了一种EBV表位高亲和力T细胞受体。本发明所提供的EBV表位高亲和力T细胞受体包含α链和β链；所述α链包含三个互补决定区,序列分别为SEQ ID No.3的第43-49位、第67-71位以及第106-116位；所述β链包含三个互补决定区,氨基酸序列分别为SEQ ID No.4的第46-50位、第68-73位以及第111-118位。本发明所提供的表达TCR的T细胞可以有效的识别T2细胞负载的EBV抗原多肽,并分泌IFN-γ,证明其具有功能。本发明提供的EBV表位高亲和力T细胞受体具有重要的应用价值。(The invention discloses an EBV epitope high-affinity T cell receptor. The EBV epitope high-affinity T cell receptor provided by the invention comprises an alpha chain and a beta chain; the alpha chain comprises three complementarity determining regions with the sequences of 43-49, 67-71 and 106-116 of SEQ ID No. 3; the beta-strand comprises three complementarity determining regions having amino acid sequences at positions 46-50, 68-73 and 111-118 of SEQ ID No.4, respectively. The T cell expressing the TCR provided by the invention can effectively recognize EBV antigen polypeptide loaded by T2 cells, and secrete IFN-gamma, so that the T cell expressing the TCR has functions. The EBV epitope high-affinity T cell receptor provided by the invention has important application value.)

The method of

1. A T cell receptor that recognizes an EBV antigen comprising an a chain and a P chain;

the a chain comprises three complementarity determining regions, and the amino acid sequences are respectively the 43 th-49 th position, the 67 th-71 th position and the 106 th-116 th position of SEQ ID number 3; or variants of these sequences having up to 3, 2 or 1 amino acid changes;

the P chain comprises three complementarity determining regions, and the amino acid sequences are respectively 46 th-50 th position, 68 th-73 th position and 111 th-118 th position of SEQ ID number 4; or variants of these sequences having up to 3, 2 or 1 amino acid changes.

2. The T cell receptor of claim 1, wherein: the amino acid sequence of the variable region of the a chain is 18 th to 127 th positions of SEQ ID number 3; or variants of these sequences having up to 3, 2 or 1 amino acid changes;

the amino acid sequence of the variable region of the P chain is 20 th to 128 th positions of SEQ ID number 4; or variants of these sequences having up to 3, 2 or 1 amino acid changes.

3. The T cell receptor of claim 1 or 2, wherein: the amino acid sequence of the constant region of the a chain is the 128-268 th position of SEQ ID number 3; the amino acid sequence of the constant region of the P chain is 129-307 of SEQ ID number 4.

4. The T cell receptor of any one of claims 1-3, wherein: the amino acid sequence of the a chain is SEQ ID number 3; the amino acid sequence of the P chain is SEQ ID number 4.

5. A nucleic acid molecule encoding the T cell receptor of any one of claims 1-4.

6. The nucleic acid molecule of claim 5, wherein: the nucleic acid molecule encoding the T cell receptor comprises a nucleic acid molecule encoding the a chain of the T cell receptor and a nucleic acid molecule encoding the P chain of the T cell receptor;

the sequences of the nucleic acid molecules encoding the three complementarity determining regions in the a chain of the T cell receptor are respectively 127-147 th, 199-213 th and 316-348 th of SEQ ID number 1; or a sequence which has 99% or more, 95% or more, 90% or more, 85% or more, or 80% or more identity to these sequences and encodes the same amino acid residue;

the sequences of the nucleic acid molecules encoding the three complementarity determining regions in the P chain of the T cell receptor are respectively the positions 136-150, 202-219 and 331-354 of SEQ ID number 2; or a sequence which has 99% or more, 95% or more, 90% or more, 85% or more, or 80% or more identity to these sequences and encodes the same amino acid residue.

7. The nucleic acid molecule of claim 5 or 6, wherein: the sequence of the nucleic acid molecule for encoding the variable region of the a chain is 52 th to 381 th positions of SEQ ID number 1; or a sequence which has 99% or more, 95% or more, 90% or more, 85% or more, or 80% or more identity to these sequences and encodes the same amino acid residue;

the sequence of the nucleic acid molecule of the variable region of the P chain is 58 th to 384 th positions of SEQ ID number 2; or a sequence which has 99% or more, 95% or more, 90% or more, 85% or more, or 80% or more identity to these sequences and encodes the same amino acid residue.

8. The nucleic acid molecule of any one of claims 5-7, wherein: the sequence of the nucleic acid molecule for coding the a chain is SEQ ID number l; the sequence of the nucleic acid molecule for coding the P chain is SEQ ID number 2.

9. An expression cassette, vector or cell comprising the nucleic acid molecule of any one of claims 5 to 8.

10. The carrier of claim 9, wherein: the vector is a recombinant plasmid obtained by inserting a nucleic acid molecule encoding the a chain of the T cell receptor and a nucleic acid molecule encoding the P chain of the T cell receptor between the multiple cloning sites of the vector pRRLSIN.

11. The cell of claim 9, wherein: the cell is a T cell.

12. T-cells having a T-cell receptor according to any one of claims 1 to 4.

13. A pharmaceutical composition comprising a vector or cell according to any one of claims 9 to 11, or a T cell according to claim 12.

14. Use of a T cell receptor according to any one of claims 1 to 4, or a nucleic acid molecule according to any one of claims 5 to 8, or a vector or cell according to any one of claims 9 to 11, or a T cell according to claim 12 for the manufacture of a medicament for the prophylaxis and/or treatment of a disease caused by EBV infection.

15. Use of a T cell receptor according to any one of claims 1 to 4, or a nucleic acid molecule according to any one of claims 5 to 8, or a vector or cell according to any one of claims 9 to 11, or a T cell according to claim 12 for the prophylaxis and/or treatment of a disease caused by EBV infection.

16. A method for preventing and/or treating a disease caused by EBV infection, comprising the steps of: use of a T cell receptor according to any one of claims 1 to 4, or a nucleic acid molecule according to any one of claims 5 to 8, or a vector or cell according to any one of claims 9 to 11, or a T cell according to claim 12 for the prevention and/or treatment of a disease caused by EBV infection.

17. The use according to claim 14 or 15 or the method according to claim 16, characterized in that: the disease caused by EBV infection is nasopharyngeal carcinoma and/or oropharyngeal squamous cell tumor and/or T cell malignancy.