Compositions and methods for immunomodulation in an organism using IL-15 and soluble IL-15Ra
The present invention relates to a therapeutic polypeptide and methods for its creation and use for modulating an immune response in a host organism in need thereof. In particular, the invention relates to the administration to an organism in need thereof, of an effective amount of a pre-coupled polypeptide complex comprising a lymphokine polypeptide portion, for example IL-15 (SEQ ID NO: 5, 6), IL-2 (SEQ ID NO: 10, 12) or combinations of both, and an interleukin receptor polypeptide portion, for example IL-15Ra (SEQ ID NO: 7, 8), IL-2Ra (SEQ ID NO: 9, 11) or combinations of both, for augmenting the immune system in, for example, cancer, SCID, AIDS, or vaccination; or inhibiting the immune system in, for example, rheumatoid arthritis, or Lupus. The therapeutic complex of the invention surprisingly demonstrates increased half-life, and efficacy in vivo.
1. A method for increasing proliferation of immune cells, comprising administering to a human an effective amount of a composition, wherein the composition comprises:
a. a purified interleukin-15 (IL-15) polypeptide and a purified soluble form of an interleukin-15 receptor alpha (IL-15Ra) polypeptide;
b. a purified chimeric polypeptide in which an IL-15 polypeptide is linked to a soluble form of an IL-15Ra polypeptide;
c. a purified IL-15 polypeptide and a purified chimeric polypeptide in which a soluble form of an IL-15Ra polypeptide is covalently linked to the Fc portion of an antibody;
d. a purified complex comprising an IL-15 polypeptide bound to a soluble form of an IL-15Ra polypeptide; or
e. a purified complex comprising an IL-15 polypeptide bound to a soluble form of an IL-15Ra polypeptide that is covalently linked to the Fc portion of an antibody,
wherein said administration results in increasing proliferation of immune cells.
2. The method of claim 1 , wherein the composition is administered by a parenteral route.
3. The method of claim 1 , wherein the composition is administered by an intravenous route, subcutaneous route or intramuscular route.
4. The method of claim 1 , wherein the human has cancer, or the human has received a bone marrow or stem cell transplant.
5. The method of claim 1 , wherein the human is immunodeficient.
6. The method of claim 1 , wherein the human has AIDS.
7. The method of claim 1 , wherein the human is lymphopenic.
8. The method of claim 1 , wherein the effective amount increases proliferation of the immune cells as measured in an in vitro assay with CFSE-labeled immune cells, wherein CFSE dilution is measured by flow cytometry.
9. The method of claim 1 , wherein the composition comprises a purified IL-15 polypeptide and a purified soluble form of an IL-15Ra polypeptide.
10. The method of claim 1 , wherein the composition comprises a purified complex comprising an IL-15 polypeptide bound to a soluble form of an IL-15Ra polypeptide.
11. The method of claim 9 or 10 , wherein the IL-15 polypeptide is:
(i) (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 2 over the entire length of SEQ ID NO: 2; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 6 over the length of the mature form of SEQ ID NO: 6; and
(ii) capable of forming a complex with IL-15Ra polypeptide.
12. The method of claim 11 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
13. The method of claim 9 or 10 , wherein the soluble form of the IL-15Ra polypeptide is:
(i) a soluble form of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) capable of forming a complex with IL-15 polypeptide.
14. The method of claim 9 or 10 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
15. The method of claim 1 , wherein the IL-15 polypeptide is:
(i) (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 2 over the entire length of SEQ ID NO: 2; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 6 over the length of the mature form of SEQ ID NO: 6; and
(ii) capable of forming a complex with IL-15Ra polypeptide.
16. The method of claim 15 , wherein the soluble form of the IL-15Ra polypeptide is:
(i) a soluble form of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) capable of forming a complex with IL-15 polypeptide.
17. The method of claim 15 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
18. The method of claim 17 , wherein the IL-15Ra polypeptide has the amino acid sequence of SEQ ID NO:8.
19. The method of claim 1 , wherein the IL-15 polypeptide has the amino acid sequence of SEQ ID NO:6.
20. The method of claim 19 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
21. The method of claim 19 or 20 , wherein the IL-15Ra polypeptide has the amino acid sequence of SEQ ID NO:8.
22. The method of claim 1 , wherein the soluble form of the IL-15Ra polypeptide is:
(i) a soluble form of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) capable of forming a complex with IL-15 polypeptide.
23. The method of claim 1 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
24. The method of claim 23 , wherein the IL-15Ra polypeptide has the amino acid sequence of SEQ ID NO:8.
25. A method for driving homeostatic proliferation of lymphokine responsive immune cells, comprising administering to a human an effective amount of a composition, wherein the composition comprises:
a. a purified interleukin-15 (IL-15) polypeptide and a purified soluble form of an interleukin-15 receptor alpha (IL-15Ra) polypeptide;
b. a purified chimeric polypeptide in which an IL-15 polypeptide is linked to a soluble form of an IL-15Ra polypeptide;
c. a purified IL-15 polypeptide and a purified chimeric polypeptide in which a soluble form of an IL-15Ra polypeptide is covalently linked to the Fc portion of an antibody;
d. a purified complex comprising an IL-15 polypeptide bound to a soluble form of an IL-15Ra polypeptide; or
e. a purified complex comprising an IL-15 polypeptide bound to a soluble form of an IL-15Ra polypeptide that is covalently linked to the Fc portion of an antibody,
wherein said administration results in driving homeostatic proliferation of lymphokine responsive immune cells.
26. The method of claim 1 or 25 , wherein said purified IL-15 or said purified complex demonstrates an in vivo half-life of greater than an hour.
27. The method of claim 25 , wherein the composition is administered by a parenteral route.
28. The method of claim 25 , wherein the composition is administered by an intravenous route, subcutaneous route or intramuscular route.
29. The method of claim 25 , wherein the human has cancer, or the human has received a bone marrow or stem cell transplant.
30. The method of claim 25 , wherein the human is immunodeficient.
31. The method of claim 25 , wherein the human has AIDS.
32. The method of claim 25 , wherein the soluble form of the IL-15Ra polypeptide is:
(i) a soluble form of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) capable of forming a complex with IL-15 polypeptide.
33. The method of claim 25 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
34. The method of claim 33 , wherein the IL-15Ra polypeptide has the amino acid sequence of SEQ ID NO:8.
35. The method of claim 25 , wherein the IL-15 polypeptide has the amino acid sequence of SEQ ID NO:6.
36. The method of claim 35 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
37. The method of claim 35 or 36 , wherein the IL-15Ra polypeptide has the amino acid sequence of SEQ ID NO:8.
38. The method of claim 25 , wherein the IL-15 polypeptide is:
(i) (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 2 over the entire length of SEQ ID NO: 2; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 6 over the length of the mature form of SEQ ID NO: 6; and
(ii) capable of forming a complex with IL-15Ra polypeptide.
39. The method of claim 38 , wherein the soluble form of the IL-15Ra polypeptide is
(i) a soluble form of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
40. The method of claim 38 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
41. The method of claim 40 , wherein the IL-15Ra polypeptide has the amino acid sequence of SEQ ID NO:8.
42. The method of claim 25 , wherein the composition comprises a purified IL-15 polypeptide and a purified soluble form of an IL-15Ra polypeptide.
43. The method of claim 25 , wherein the composition comprises a purified complex comprising an IL-15 polypeptide bound to a soluble form of an IL-15Ra polypeptide.
44. The method of claim 42 or 43 , wherein the IL-15 polypeptide is:
(i) (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 2 over the entire length of SEQ ID NO: 2; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 6 over the length of the mature form of SEQ ID NO: 6; and
(ii) capable of forming a complex with IL-15Ra polypeptide.
45. The method of claim 44 , wherein the soluble form of the IL-15Ra polypeptide:
(i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.
46. The method of claim 42 or 43 , wherein the soluble form of the IL-15Ra polypeptide is:
(i) a soluble form of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) capable of forming a complex with IL-15 polypeptide.
47. The method of claim 42 or 43 , wherein the soluble form of the IL-15Ra polypeptide: (i) has the amino acid sequence of the extracellular domain of an IL-15Ra polypeptide and the IL-15Ra polypeptide is (a) encoded by a nucleic acid that is at least 80% identical to SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; or (b) at least 95% identical to the amino acid sequence of SEQ ID NO: 8 over the length of the mature form of SEQ ID NO: 8; and
(ii) is capable of forming a complex with IL-15 polypeptide.