Compositions and methods for immunomodulation in an organism

Patent metadata

Field	Value
Publication number	US-10464993-B2
Application number	US-201815946402-A
Country	US
Kind code	B2
Filing date	Apr 5, 2018
Priority date	May 17, 2005
Publication date	Nov 5, 2019
Grant date	Nov 5, 2019

Abstract

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.

First claim

We claim:

1. A nucleic acid encoding an interleukin polypeptide complex comprising an interleukin polynucleotide with at least 95% homology to a nucleic acid set forth in SEQ ID NO: 2, and an interleukin receptor polynucleotide that has at least 95% homology to a nucleic acid set forth in SEQ ID NO: 4, wherein the interleukin polynucleotide and interleukin receptor polynucleotide are capable of being expressed as a single polypeptide.

2. The nucleic acid of claim 1 , further comprising a polynucleotide segment encoding an antibody Fc portion.

3. An isolated host cell containing the nucleic acid molecules of claim 1 or claim 2 .

4. An expression vector comprising the nucleic acid of claim 1 .

5. The expression vector of claim 4 , further comprising a transcription regulator sequence, fusion protein sequences, linker sequences, or any combination thereof.

6. The expression vector of claim 4 , comprising a transcription regulator sequence that is a promoter, inducible promoter, enhancer, or any combination thereof.

7. The expression vector of claim 5 , comprising a fusion protein sequence that is a His-tag, GST, GFP, antibody Fc portion, antibiotic resistance, signal peptides, or any combination thereof.

8. The expression vector of claim 5 , comprising a linker sequence that is disposed at the 5′ end, 3′ end, a location within the polypeptide encoding sequences, or any combination thereof.

9. The nucleic acid of claim 1 , wherein the nucleic acid is disposed in a viral vector, bacterial plasmid, or artificial chromosome.

10. A method of making a polypeptide complex, comprising:
a) culturing a host cell comprising a plurality of nucleic acids encoding an interleukin polypeptide complex comprising an interleukin polynucleotide with at least 95% homology to a nucleic acid set forth in SEQ ID NO: 2, and an interleukin receptor polynucleotide that has at least 95% homology to a nucleic acid set forth in SEQ ID NO: 4;
b) isolating an interleukin polypeptide expressed by the interleukin polynucleotide;
c) isolating an interleukin receptor polypeptide expressed by the interleukin receptor polynucleotide; and
d) contacting the interleukin polypeptide and the interleukin receptor polypeptide for from 1 minute to 120 minutes, at from 26° C. to 40° C., in a suitable buffer having a pH from 5.5 to 8.5.

11. The method of claim 10 , wherein the host cell is a mammalian cell.

12. The method of claim 11 , wherein the mammalian cell is a Chinese hamster ovary cell.

13. The method of claim 10 , wherein the interleukin polynucleotide and/or interleukin receptor polynucleotide further comprises a polynucleotide segment encoding an antibody Fc portion.

14. The method of claim 10 , wherein the interleukin polynucleotide and/or interleukin receptor polynucleotide further comprises a transcription regulator sequence, fusion protein sequences, linker sequences, or any combination thereof.

15. The method of claim 14 , comprising a transcription regulator sequence that is a promoter, inducible promoter, enhancer, or any combination thereof.

16. The method of claim 14 , comprising a fusion protein sequence that is a His-tag, GST, GFP, antibody Fc portion, antibiotic resistance, signal peptides, or any combination thereof.

17. The method of claim 14 , comprising a linker sequence that is disposed at the 5′ end, 3′ end, a location within the polypeptide encoding sequences, or any combination thereof.

18. The method of claim 10 , wherein plurality of nucleic acids is disposed in a viral vector, bacterial plasmid, or artificial chromosome.

19. The method of claim 10 , further comprising isolating the product from d).

Assignees

• Univ Connecticut

Inventors

• Lefrancois Leo
• Stoklasek Thomas A

Classifications

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P37/04

  Immunostimulants · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• A61P35/02

  specific for leukemia · CPC title