Gene therapy with interleukin-10 receptor and interleukin-12 induces a protective interferon-γ-dependent response against B16F10-Nex2 melanoma.

Abstract

Antitumor immune responses are associated with proinflammatory cytokines, whereas tumor-developing animals generally have increased the production of immunosuppressive cytokines. Here, we show that splenocytes from C57Bl/6 mice resistant to low doses of B16F10-Nex2 melanoma cells produced twofold or higher interferon-γ (IFN-γ)/interleukin-10 (IL-10) ratios, whereas cells from tumor-bearing animals produced predominantly IL-10. IL-10-knockout (IL-10KO) mice were significantly more resistant to B16F10-Nex2 development, producing increased amounts of IL-12 and IFN-γ. To neutralize IL-10 in vivo, aiming at cancer therapy, recombinant eukaryotic plasmid expressing the soluble extracellular region of the murine IL-10 receptor α-chain was constructed (pcDNA3-sIL-10R). Plasmid-treated melanoma-challenged animals showed extended survival time, the protective response was IFN-γ dependent and enhanced by co-immunization with a plasmid expressing IL-12. Dendritic cells (DCs) from IL-10KO mice, primed with B16F10-Nex2 antigens (TAg), secreted increased amounts of T-helper 1-type cytokines and increased the expression of surface activation markers. Vaccination of C57Bl/6 mice with TAg-activated IL-10KO DCs, as well as with TAg-primed DCs from C57Bl/6 mice transfected with pcDNA3-sIL10R plasmid, significantly increased animal survival. In conclusion, an IFN-γ-dependent protective response was induced against B16F10-Nex2 cells by neutralization of IL-10 with pcDNA3-sIL10R plasmid. This effect was enhanced by association with IL-12 gene therapy (80% protection), and could be mediated by TAg-primed DCs.

• Authors: L H L Marchi, T Paschoalin, L R Travassos, E G Rodrigues
• Journal: Cancer gene therapy (Cancer Gene Ther) Vol. 18 Issue 2 Pg. 110-22 (Feb 2011) ISSN: 1476-5500 [Electronic] England
• PMID: 20885448 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Receptors, Interleukin-10
• Interleukin-12
• Interferon-gamma

Topics

• Animals
• Cell Line, Tumor
• Dendritic Cells (immunology)
• Genetic Therapy (methods)
• Humans
• Immunotherapy, Adoptive (methods)
• Interferon-gamma (immunology)
• Interleukin-12 (biosynthesis, genetics, immunology)
• Male
• Melanoma, Experimental (genetics, immunology, therapy)
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Plasmids (genetics)
• Receptors, Interleukin-10 (biosynthesis, genetics, immunology)
• Transfection