Cells expressing a modified T cell receptor
View Patent ↗This invention provides a cell presenting at least one T cell receptor (TCR) anchored to the membrane by a transmembrane sequence, said TCR comprising an interchain disulfide bond between extracellular constant domain residues which is not present in native TCRs.
1. A mammalian cell comprising a membrane presenting at least one modified T cell receptor (TCR) anchored to the membrane by a transmembrane sequence, wherein the modified TCR comprises an interchain disulfide bond between extracellular constant domain residues which is not present in native TCRs.
2. A mammalian cell comprising a membrane, presenting at least one modified αβ T cell receptor (TCR) anchored to the membrane by a transmembrane sequence, wherein the modified αβ TCR comprises a disulfide bond between α and β extracellular constant domain residues which is not present in native TCRs.
3. A cell as claimed in claim 1 wherein the modified TCR is an αβ heterodimeric TCR.
4. A cell as claimed in claim 3 wherein the modified TCR comprises α and β chains and wherein each chain comprises a transmembrane sequence, fused at its N terminus to an extracellular constant domain sequence, in turn fused at its N terminus to a variable region sequence.
5. A cell as claimed in claim 4 wherein at least sequences of the modified TCR α and β chains, other than complementarity determining regions of the variable region, correspond to human TCR α and β sequences.
6. A cell as claimed in claim 1 wherein the modified TCR is an αβ single chain TCR.
7. A cell as claimed in claim 6 wherein the modified TCR comprises:
(i) a first segment constituted by an α chain variable region sequence fused to the N terminus of an α chain extracellular constant domain sequence, and a second segment constituted by a β chain variable region sequence fused to the N terminus of a sequence β chain extracellular constant and transmembrane sequences, and a linker sequence linking the C terminus of the first segment to the N terminus of the second segment, or
(ii) a first segment constituted by a TCR β chain variable region sequence fused to the N terminus of a β chain extracellular constant domain sequence, and a second segment constituted by an α chain variable region sequence fused to the N terminus of a sequence α chain extracellular constant and transmembrane sequences, and a linker sequence linking the C terminus of the first segment to the N terminus of the second segment.
8. A cell as claimed in claim 7 wherein at least the sequences of the modified TCR α and β chains, other than complementarity determining regions of the variable region, correspond to human TCR α and β sequences.
9. A cell as claimed in claim 1 displaying a plurality of modified TCRs.
10. A cell as claimed in claim 1 wherein the cell is a T cell.
11. A cell as claimed in claim 1 wherein the cell is a cytotoxic T cell.
12. A cell as claimed in claim 1 wherein the cell reduces cellular or proinflammatory arms of an auto-immune response.
13. A cell as claimed in claim 12 wherein the cell is a regulatory T cell.
14. A cell as claimed in claim 1 wherein in the modified TCR a covalent disulfide bond links a residue of an immunoglobulin region of a constant domain of the α chain to a residue of an immunoglobulin region of a constant domain of the β chain.
15. A cell as claimed in claim 1 wherein the modified TCR has no equivalent of the interchain disulfide bond present in native TCRs.
16. A cell as claimed in claim 15 wherein in the modified TCR the cysteine residues which form the interchain disulfide bond present in native TCRs are replaced by noncysteine residues.
17. A cell as claimed in claim 15 , wherein in the modified TCR cysteine residues which form the interchain disulfide bond present in native TCRs are replaced by serine or alanine.
18. A cell as claimed in claim 1 wherein in the modified TCR, an unpaired cysteine residue present in native TCR β chains is not present.
19. A cell as claimed in claim 1 wherein in the modified TCR the disulfide bond is between cysteine residues substituted for residues whose β carbon atoms are less than 0.6 nm apart in the native TCR structure.
20. A cell as claimed in claim 1 wherein in the modified TCR the disulfide bond is between cysteine residues substituted for Thr 48 of exon 1 of TRAC*01 and Ser 57 of exon 1 of TRBC1*01 or TRBC2*01.
21. A cell as claimed in claim 1 , wherein in the modified TCR the disulfide bond is between cysteine residues substituted for Thr 45 of exon 1 of TRAC*01 and Ser 77 of exon 1 of TRBC1*01 or TRBC2*01.
22. A cell as claimed in claim 1 , wherein in the modified TCR the disulfide bond is between cysteine residues substituted for Tyr 10 of exon 1 of TRAC*01 and Ser 17 of exon 1 of TRBC1*01 or TRBC2*01.
23. A cell as claimed in claim 1 , wherein in the modified TCR the disulfide bond is between cysteine residues substituted for Thr 45 of exon 1 of TRAC*01 and Asp 59 of exon 1 of TRBC1*01 or TRBC2*01.
24. A cell as claimed in claim 1 , wherein in the modified TCR the disulfide bond is between cysteine residues substituted for Ser 15 of exon 1 of TRAC*01 and Glu 15 of exon 1 of TRBC1*01 or TRBC2*01.
25. An infusible or injectable pharmaceutical composition comprising a plurality of T cells as claimed in claim 1 together with a pharmaceutically acceptable carrier.