Biologically relevant orthogonal cytokine/receptor pairs
Engineered orthogonal cytokine receptor/ligand pairs, and methods of use thereof, are provided.
1. A nucleic acid encoding an engineered human IL-2 polypeptide, which comprises amino acid substitutions, numbered relative to reference SEQ ID NO:4:
E15S, E15T, E15Q, or E15H;
H16Q;
L19V or L19I;
D20T, D20S, D20L or D20M; and
M23L, M23S, M23V, M23A, or M23T;
and optionally comprises Q22K, Q22N, R81D, R81Y, T51I or a combination thereof.
2. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises amino acid substitution Q22K.
3. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises amino acid substitution T51I.
4. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises amino acid substitution R81D or R81Y.
5. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises amino acid substitutions Q22K and M23A.
6. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises amino acid substitutions M23Q, R81D and T51I.
7. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises amino acid substitutions M23Q and R81Y.
8. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises amino acid substitution M23A.
9. A nucleic acid encoding an engineered human IL-2 polypeptide, which comprises a set of amino acid substitutions, numbered relative to reference SEQ ID NO:4:
E15S;
H16Q;
L19V;
D20L;
M23A; and
Q22K.
10. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises a set of amino acid substitutions, numbered relative to reference SEQ ID NO:4:
E15S;
H16Q;
L19V;
D20L;
M23Q;
R81D; and
T51I.
11. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises a set of amino acid substitutions, numbered relative to reference SEQ ID NO:4:
E15S;
H16Q;
L19V;
D20L;
M23Q;
and R81Y.
12. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide comprises a set of amino acid substitutions, numbered relative to reference SEQ ID NO:4:
E15S;
H16Q;
L19V;
D20L; and
M23A.
13. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide is fused to an Fc domain of IgG, or albumin.
14. The nucleic acid of claim 1 , wherein the human IL-2 polypeptide binds to and activates an orthogonal human CD122 protein.
15. The nucleic acid of claim 1 , wherein the nucleic acid is RNA.
16. The nucleic acid of claim 1 , wherein the nucleic acid is DNA.
17. An expression vector comprising the nucleic acid of claim 1 .
18. The expression vector of claim 17 , wherein the vector is a viral vector.
19. The expression vector of claim 17 , wherein the vector is a plasmid vector.
20. The expression vector of claim 17 , wherein the vector is an integrating vector.
21. A cell genetically engineered to comprise the nucleic acid of claim 1 .
22. The nucleic acid of claim 9 , wherein the human IL-2 polypeptide binds to and activates an orthogonal human CD122 protein.
23. The nucleic acid of claim 22 , wherein the nucleic acid is RNA.
24. The nucleic acid of claim 22 , wherein the nucleic acid is DNA.
25. An expression vector comprising the nucleic acid of claim 22 .
26. The expression vector of claim 25 , wherein the vector is a viral vector.
27. The expression vector of claim 25 , wherein the vector is a plasmid vector.
28. The expression vector of claim 25 , wherein the vector is an integrating vector.
29. A cell genetically engineered to comprise the nucleic acid of claim 9 .