Compositions and methods for immunomodulation in an organism
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 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).