Complexes of IL-15 and IL-15Ralpha and uses thereof
The present invention relates to agents that modulate interleυkin-15 (“IL-15”) signal transduction or function (“Therapeutic Agents”) and the use ol” those agents to modulate immune function. The Therapeutic Agents target the interaction between IL-15 and its receptor and modulate IL-15-induced signal transduction. The Therapeutic Agents may be formulated with polymers, such as poly-β-1-♦4-N-acetylglucosamine. for administration to a human subject to modulate IL-15-mediated immune function.
1. A method of recombinantly producing a codon optimized human IL-15 and a codon optimized human IL-15Ra complex, comprising:
(a) culturing a cell expressing a human IL-15 encoded by:
(i) a nucleic acid sequence with at least 95% identity to nucleic acid residues 769 to 1257 of SEQ ID NO:9;
(ii) a nucleic acid sequence with at least 95% identity to nucleic acid residues 769 to 1218 of SEQ ID NO:11; or
(iii) a nucleic acid sequence with at least 95% identity to nucleic acid residues 769 to 1164 of SEQ ID NO:17;
and a human IL15Ra encoded by:
(iv) a nucleic acid sequence with at least 95% identity to nucleic acid residues 775 to 1581 of SEQ ID NO:13; or
(v) a nucleic acid sequence with at least 95% identity to nucleic acid residues 775 to 1389 of SEQ ID NO:15; and
(b) purifying the human IL-15 and human IL-15Ra complex, wherein the cell expresses at least 0.15 pg of human IL-15 per day when cultured in serum-free media as measured by an ELISA.
2. The method of claim 1 , wherein the human IL-15 is encoded by the nucleic acid sequence comprising nucleic acid residues 769 to 1257 of SEQ ID NO:9; and the human IL15Ra is encoded by the nucleic acid sequence comprising nucleic acid residues 775 to 1581 of SEQ ID NO:13.
3. The method of claim 1 , wherein the human IL-15 is encoded by the nucleic acid sequence comprising nucleic acid residues 769 to 1257 of SEQ ID NO:9; and the human IL15Ra is encoded by the nucleic acid sequence comprising nucleic acid residues 775 to 1389 of SEQ ID NO:15.
4. The method of claim 1 , wherein the human IL-15 is encoded by the nucleic acid sequence comprising nucleic acid residues 769 to 1218 of SEQ ID NO:11; and the human IL15Ra is encoded by the nucleic acid sequence comprising nucleic acid residues 775 to 1581 of SEQ ID NO:13.
5. The method of claim 1 , wherein the human IL-15 is encoded by the nucleic acid sequence comprising nucleic acid residues 769 to 1218 of SEQ ID NO:11; and the human IL15Ra is encoded by the nucleic acid sequence comprising nucleic acid residues 775 to 1389 of SEQ ID NO:15.
6. The method of claim 1 , wherein the human IL-15 is encoded by the nucleic acid sequence comprising nucleic acid residues 769 to 1164 of SEQ ID NO:17; and the human IL15Ra is encoded by the nucleic acid sequence comprising nucleic acid residues 775 to 1581 of SEQ ID NO:13.
7. The method of claim 1 , wherein the human IL-15 is encoded by the nucleic acid sequence comprising nucleic acid residues 769 to 1164 of SEQ ID NO:17; and the human IL15Ra is encoded by the nucleic acid sequence comprising nucleic acid residues 775 to 1389 of SEQ ID NO:15.
8. The method of claim 1 , wherein the cell is selected from the group consisting of: 293, COS, CHO, HeLa, NIH3T3, HepG2, MCF7, RD, PC12, K562, skBr3, BT274, A204, M07Sb, TFB1, Raji, Jurkat, MOLT-4, CTLL-2, MC-IXC, SK-N-MC, SK-N-DZ, SH-SYS5Y, C127 and BE(2)-C cell line.
9. The method of claim 8 , wherein the cell is from the 293 or CHO cell line.
10. The method of claim 1 , wherein the cell expresses at least 0.6 pg of human IL-15 per day.
11. The method of claim 1 , wherein the cell expresses at least 1 pg of human IL-15 per day.
12. The method of claim 1 , wherein the cell expresses up to 2 pg of human IL-15 per day.