High-affinity DMF5 T cell receptor (TCR) variants
High-affinity variants of the DMF5 TCR, and methods of use thereof for the treatment and imaging of cancer in a patient.
1. A high-affinity DMF5 T cell receptor variant α chain, wherein the aspartic amino acid at position 27 or 49 is a Y (αD27Y or αD49Y) or W (αD27W or αD49W).
2. A high-affinity DMF5 T cell receptor variant β chain, wherein the leucine amino acid at position 95, 96 or 114 is a W (βL95W, βL96W βL114W).
3. A high-affinity DMF5 T cell receptor variant comprising:
a high-affinity DMF5 T cell receptor variant α chain, wherein the aspartic amino acid at
position 27 or 49 is a Y (αD27Y or αD49Y) or W (αD27W or αD49W), and
a high-affinity DMF5 T cell receptor variant β chain, wherein the leucine amino acid at position 95, 96 or 114 is a W (βL95W, βL96W βL114W).
4. A fusion protein comprising the high-affinity DMF5 T cell receptor variant of claim 3 linked to a non-TCR sequence.
5. A therapeutic composition comprising:
a therapeutic agent linked to
the high-affinity DMF5 T cell receptor variant of claim 3 , optionally linked to a non-TCR sequence.
6. The therapeutic composition of claim 5 , wherein the therapeutic agent is a radioisotope; an anticancer drug; or a cytotoxic moiety.
7. A composition comprising:
a detectable moiety linked to
the high-affinity DMF5 T cell receptor variant of claim 3 , optionally linked to a non-TCR sequence.
8. The composition of claim 7 , wherein the detectable moiety is a contrast agent.
9. The composition of claim 8 , wherein the contrast agent is selected from the group consisting of gold, gadolinium, iron oxides, manganese chelates, barium sulfate, iodinated contrast media, microbubbles, and perfluorocarbons.
10. A nucleic acid encoding:
the high-affinity DMF5 T cell receptor variant of claim 3 , optionally linked to a non-TCR sequence.
11. A vector comprising the nucleic acid of claim 10 .
12. The vector of claim 11 , which is a retroviral vector.
13. A host cell expressing:
the high-affinity DMF5 T cell receptor variant of claim 3 , optionally linked to a non-TCR sequence.
14. A host cell comprising the nucleic acid of claim 10 .
15. The host cell of claim 14 , wherein the cell is a peripheral blood lymphocyte (PBL).
16. A method of treating a subject who has melanoma, the method comprising administering to the subject a therapeutically effective amount of the therapeutic composition of claim 5 .
17. A method of treating a subject who has melanoma, the method comprising administering to the subject a therapeutically effective amount of the host cells of claim 14 .
18. The method of claim 17 , wherein the host cells are autologous to the subject.
19. A method of treating a subject who has melanoma, the method comprising:
obtaining cells from the subject;
transducing the cells with a vector expressing:
the high-affinity DMF5 T cell receptor variant of claim 3 , optionally linked to a non-TCR sequence;
optionally maintaining the cells in culture for a time sufficient for the cells to express the variant or fusion protein and/or to proliferate; and
administering a therapeutically effective amount of the cells to the subject thereafter.
20. The method of claim 19 , wherein the cells are PBLs.