Multiple cytokine protein complexes
View Patent ↗The invention relates to protein complexes and fusion proteins including at least two different cytokine molecules. The protein complexes and fusion proteins may further include a targeting moiety such as a region of an immunoglobulin. Methods of using the protein complexes and fusion proteins are also disclosed.
1. A fusion protein comprising:
a first polypeptide chain comprising the p35 subunit of interleukin-12 (IL-12); and
a second polypeptide chain comprising a cytokine and the p40 subunit of IL-12, wherein the cytokine is a four-helix bundle protein.
2. The fusion protein of claim 1 , wherein the first polypeptide chain is covalently bonded to the second polypeptide chain.
3. The fusion protein of claim 2 , wherein the first polypeptide chain is covalently bonded to the second polypeptide chain by a disulfide bond.
4. The fusion protein of claim 1 , wherein the cytokine is interleukin-2 (IL-2).
5. The fusion protein of claim 1 , wherein the cytokine is granulocyte-macrophage colony stimulating factor (GM-CSF).
6. The fusion protein of claim 1 , wherein the cytokine is selected from the group consisting of interleukin-4 (IL-4), interleukin-6 (IL-6), and macrophage colony-stimulating factor (M-CSF).
7. A nucleic acid encoding the fusion protein of claim 1 .
8. An isolated or cultured cell comprising the nucleic acid of claim 7 .
9. A nucleic acid encoding the fusion protein of claim 4 .
10. An isolated or cultured cell comprising the nucleic acid of claim 9 .
11. A fusion protein comprising a single-chain polypeptide comprising:
the p35 subunit of interleukin-12 (IL-12);
the p40 subunit of IL-12; and
a second cytokine, wherein the second cytokine is a four-helix bundle protein, and wherein the p35 subunit of IL-12, the p40 subunit of IL-12, and the second cytokine are fused together as portions of a single, expressed polypeptide chain.
12. The fusion protein of claim 11 , wherein the second cytokine is interleukin-2 (IL-2).
13. The fusion protein of claim 11 , wherein the second cytokine is granulocyte-macrophage colony stimulating factor (GM-CSF).
14. The fusion protein of claim 11 , wherein the second cytokine is selected from the group consisting of interleukin-4 (IL-4), interleukin-6 (IL-6), and macrophage colony-stimulating factor (M CSF).
15. A nucleic acid encoding the fusion protein of claim 11 .
16. An isolated or cultured cell comprising the nucleic acid of claim 15 .
17. The fusion protein of claim 11 comprising, in an N- to C-terminal direction, the p35 subunit of IL-12, the p40 subunit of IL-12, and the second cytokine.
18. A nucleic acid encoding the fusion protein of claim 12 .
19. An isolated or cultured cell comprising the nucleic acid of claim 18 .
20. A fusion protein comprising:
a first polypeptide chain comprising the p40 subunit of interleukin-12 (IL-12); and
a second polypeptide chain comprising a cytokine and the p35 subunit of IL-12, wherein the cytokine is a four-helix bundle protein.
21. The fusion protein of claim 20 , wherein the first polypeptide chain is covalently bonded to the second polypeptide chain.
22. The fusion protein of claim 21 , wherein the first polypeptide chain is covalently bonded to the second polypeptide chain by a disulfide bond.
23. The fusion protein of claim 20 , wherein the cytokine is interleukin-2 (IL-2).
24. The fusion protein of claim 20 , wherein the cytokine is granulocyte-macrophage colony stimulating factor (GM-CSF).
25. The fusion protein of claim 20 , wherein the cytokine is selected from the group consisting of interleukin-4 (IL-4), interleukin-6 (IL-6), and macrophage colony-stimulating factor (M-CSF).
26. A nucleic acid encoding the fusion protein of claim 20 .
27. An isolated or cultured cell comprising the nucleic acid of claim 26 .
28. A nucleic acid encoding the fusion protein of claim 23 .
29. An isolated or cultured cell comprising the nucleic acid of claim 28 .