Biologically relevant orthogonal cytokine/receptor pairs
Engineered orthogonal cytokine receptor/ligand pairs, and methods of use thereof, are provided.
1. A nucleic acid encoding a variant human CD122 polypeptide, the variant human CD122 polypeptide comprising amino acid substitutions at positions H133 and Y134, wherein numbering is made in reference to SEQ ID NO: 1.
2. An expression vector comprising the nucleic acid of claim 1 .
3. A cell genetically engineered to comprise the vector of claim 2 .
4. The nucleic acid of claim 1 wherein the variant human CD122 polypeptide further comprises an amino acid substitution or deletion at one or more residues selected from R41, R42, Q70, K71, T73, T74, V75, S132, F135, E136, and Q214.
5. The nucleic acid of claim 4 wherein the variant human CD122 polypeptide further comprises an amino acid substitution or deletion at one or more residues selected from Q70, and T73.
6. The nucleic acid of claim 4 wherein the variant human CD122 polypeptide further comprises an amino acid substitution at one or more residues selected from Q70, and T73.
7. The nucleic acid of claim 6 wherein the variant human CD122 polypeptide comprises one or more amino acid substitutions selected from 070Y, T73 D, T73Y, H133D, H133E, H133K, Y134F, Y134E, and Y134R.
8. The nucleic acid of claim 1 wherein the variant human CD122 polypeptide comprises amino acid substitutions at position 133 selected from H133D, H133E, and H133K, and at position 134 selected from Y134F, Y134E, and Y134R.
9. The nucleic acid of claim 8 wherein the variant human CD122 polypeptide comprises amino acid substitutions H133D and Y134F.
10. The nucleic acid of claim 1 further comprising a nucleic acid encoding a signal sequence, the nucleic acid encoding the signal sequence operably linked to the nucleic acid encoding the variant human CD122 polypeptide.
11. The nucleic acid of claim 10 wherein the signal sequence is the naturally occurring human CD122 signal sequence.
12. The nucleic acid of claim 11 wherein the variant human CD122 polypeptide comprises amino acid substitutions at position 133 selected from H1 33D, H1 33E, and H133K, and at position 134 selected from Y134F, Y134E, and Y134R.
13. The nucleic acid of claim 12 wherein the variant human CD122 polypeptide comprises the amino acid substitutions H133D and Y134F.
14. The expression vector of claim 2 wherein the expression vector is a plasmid or viral vector.
15. The expression vector of claim 14 wherein the expression vector is a viral vector.
16. The expression vector of claim 15 , wherein the expression vector is a retroviral vector or lentiviral vector.
17. The expression vector of claim 14 , wherein the variant human CD122 polypeptide further comprises an amino acid substitution or deletion at one or more residues selected from R41, R42, Q70, K71, T73, T74, V75, S132, F135, E136, and Q214.
18. The expression vector of claim 14 wherein the variant human CD122 polypeptide comprises an amino acid substitution or deletion at one or more residues selected from Q70, T73, H133 and Y134.
19. The expression vector of claim 18 wherein the variant human CD122 polypeptide further comprises one or more amino acid substitutions selected from Q70Y, T73D, and T73Y.
20. The expression vector of claim 19 wherein the variant human CD122 polypeptide comprises amino acid substitutions at position 133 selected from H133D, H133E, and H133K, and at position 134 selected from Y134F, Y134E, and Y134R.
21. The expression vector of claim 20 wherein the vector is a lentiviral vector or a retroviral vector.
22. The expression vector of claim 20 wherein the variant human CD122 polypeptide comprises amino acid substitutions H133D and Y134F.
23. The expression vector of claim 14 further comprising a nucleic acid encoding a signal sequence operably linked to the nucleic acid encoding the variant human CD122 polypeptide.
24. The expression vector of claim 23 wherein the signal sequence is the naturally occurring human CD122 signal sequence.
25. The expression vector of claim 23 wherein the variant human CD122 polypeptide comprises amino acid substitutions at position 133 selected from H133D, H133E, and H133K, and at position 134 selected from Y134F, Y134E, and Y134R.
26. The expression vector of claim 23 wherein the variant human CD122 polypeptide comprises the amino acid substitutions H133D and Y134F.
27. The expression vector of claim 26 wherein the vector is a viral vector.
28. The expression vector of claim 26 wherein the vector is lentiviral vector or retroviral vector.
29. The expression vector of claim 28 wherein the vector is a lentiviral vector.
30. The expression vector of claim 25 wherein the vector further comprises a nucleic acid encoding a second heterologous polypeptide.
31. The expression vector of claim 30 wherein first and second polypeptide are operably linked to a single promoter.
32. The genetically engineered cell of claim 3 , wherein the cell is a mammalian cell.
33. The genetically engineered cell of claim 32 , wherein the cell is an immune cell.
34. The genetically engineered cell of claim 33 , wherein the immune cell is a T cell.
35. The genetically engineered cell of claim 34 , wherein the T cell is selected from CD8+ T cells, CD4+ T cells, helper T cells, regulatory T cells, memory T cells, NKT cells, γδT cells.
36. The genetically engineered cell of claim 33 wherein the expression vector is a viral vector comprising a nucleic acid encoding a variant human CD122 polypeptide wherein the variant human CD122 polypeptide comprises an amino acid substitution or deletion at one or more residues selected from R41, R42, Q70, K71, T73, T74, V75, S132, F135, E136, and Q214.
37. The genetically engineered cell of claim 36 wherein the variant human CD122 polypeptide comprises an amino acid substitution or deletion at one or more residues selected from Q70 and T73.
38. The genetically engineered cell of claim 37 wherein the variant human CD122 polypeptide comprises one or more amino acid substitutions selected from Q70Y, T73D, T73Y, amino acid substitutions at position 133 selected from H133D, H133E, and H133K, and at position 134 selected from Y1 34F, Y1 34E, and Y1 34R.
39. The genetically engineered cell of claim 34 wherein the variant human CD122 polypeptide comprises an amino acid substitution or deletion at one or more residues selected from H133 and Y134.
40. The genetically engineered cell of claim 39 wherein the variant human CD122 polypeptide comprises amino acid substitutions at position 133 selected from H133D, H133E, and H133K, and at position 134 selected from Y134F, Y134E, and Y134R.
41. The genetically engineered cell of claim 39 wherein the variant human CD122 polypeptide comprises amino acid substitutions at positions H133 and Y134.
42. The genetically engineered cell of claim 41 wherein the variant human CD122 polypeptide comprises one or more amino acid substitutions selected from selected from H133D and Y134F.
43. The genetically engineered cell of claim 41 further comprising a nucleic acid encoding a signal sequence operably linked to the nucleic acid encoding the variant human CD122 polypeptide.
44. The genetically engineered cell of claim 43 wherein the signal sequence is the naturally occurring human CD122 signal sequence.
45. The genetically engineered cell of claim 41 wherein the vector is a lentiviral vector or a retroviral vector.
46. The genetically engineered cell of claim 45 wherein the vector is a lentiviral vector.
47. The genetically engineered cell of claim 46 further comprising a nucleic acid encoding a signal sequence operably linked to the nucleic acid encoding the variant human CD122 polypeptide.
48. The genetically engineered cell of claim 47 wherein the signal sequence is the naturally occurring human CD122 signal sequence.
49. The genetically engineered cell of claim 48 wherein the vector is a lentiviral vector or a retroviral vector.
50. The genetically engineered cell of claim 49 wherein the vector is a lentiviral vector.
51. The genetically engineered cell of claim 50 wherein the vector further comprises a nucleic acid encoding a second heterologous polypeptide.
52. The genetically engineered cell of claim 51 wherein first and second polypeptide are operably linked to a single promoter.