Repertoire of allo-restricted peptide-specific T cell receptor sequences and use thereof
The present invention is directed to a kit-of-parts or composition containing nucleic acid sequences coding for high-avidity, allo-restricted TCR, wherein the TCR are independently directed against the tyrosinase antigen, the melan-A antigen and the survivin antigen. The invention is further directed to a kit-of-parts or composition containing at least three groups of transgenic lymphocytes transformed with vectors coding for TCR against said antigens. Furthermore, the present invention provides a pharmaceutical composition and its use in the treatment of diseases involving malignant cells expressing said tumor-associated antigens. The invention further relates to a nucleic acid molecule coding for a TCR that recognizes the survivin antigen, a TCR encoded thereby and a T cell expressing said TCR. Further, the invention discloses a vector, a cell and a pharmaceutical composition encoding/containing same and their use in the treatment of diseases involving malignant cells expressing survivin.
1. A nucleic acid comprising the nucleotide sequence of SEQ ID NO: 79.
2. A fusion protein, comprising:
a) at least one epitope-tag, and
b) an α chain of a TCR encoded by the nucleotide sequence of SEQ ID NO:79,
wherein said epitope-tag is selected from the group consisting of:
i) an epitope-tag added to the N- and/or C-terminus of said α chain, or added into the α chain sequence, but outside the CDR3 region,
ii) an epitope-tag inserted into a constant region of said α chain, and
iii) an epitope-tag replacing a number of amino acids in a constant region of said α chain.
3. The nucleic acid of claim 1 , further comprising a second nucleotide sequence, wherein the second nucleotide sequence is SEQ ID NO:80.
4. The fusion protein of claim 2 , wherein said epitope-tag is added to the N- and/or C-terminus of said α chain.
5. The fusion protein of claim 2 , further comprising a β chain of a TCR encoded by the nucleotide sequence of SEQ ID NO:80.
6. The fusion protein of claim 5 , wherein said fusion protein further comprises at least one epitope-tag selected from the group consisting of:
i) an epitope-tag added to the N- and/or C-terminus of said β chain, or added into the β chain sequence, but outside the CDR3 region,
ii) an epitope-tag inserted into a constant region of said β chain, and
iii) an epitope-tag replacing a number of amino acids in a constant region of said β chain.
7. The fusion protein of claim 6 , wherein said epitope-tag is added to the N- and/or C-terminus of said β chain.
8. A vector comprising the nucleic acid of claim 1 .
9. A vector comprising the nucleic acid of claim 3 .
10. A cell that has been transformed with the vector of claim 8 .
11. A cell that has been transformed with the vector of claim 9 .
12. A composition comprising a fusion protein and a pharmaceutically acceptable carrier, the fusion protein comprising:
a) at least one epitope-tag, and
b) an α chain of a TCR encoded by the nucleotide sequence of SEQ ID NO:79,
wherein said epitope-tag is selected from the group consisting of:
i) an epitope-tag added to the N- and/or C-terminus of said α chain, or added into the α chain sequence, but outside the CDR3 region,
ii) an epitope-tag inserted into a constant region of said α chain, and
iii) an epitope-tag replacing a number of amino acids in a constant region of said α chain.
13. The composition of claim 12 , wherein the fusion protein further comprises a β chain of a TCR encoded by the nucleotide sequence of SEQ ID NO:80.