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

Cancer Gene Therapy volume 18, pages 110–122 (2011)Cite this article

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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.

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Acknowledgements

This work was supported by Grant 06/50634-2 from the Fundação de Amparo à Pesquisa do Estado de São Paulo, FAPESP. LHLM was a post-graduate fellow of the Brazilian National Research Council (CNPq). TP is a Post-Doctorate fellow of FAPESP and EGR and LRT are career researchers of CNPq.

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  1. Department of Microbiology, Experimental Oncology Unit, Immunology and Parasitology, Universidade Federal de São Paulo-Escola Paulista de Medicina, São Paulo, Brazil

    L H L Marchi, T Paschoalin, L R Travassos & E G Rodrigues

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  1. L H L Marchi
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  2. T Paschoalin
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Correspondence to E G Rodrigues.

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Marchi, L., Paschoalin, T., Travassos, L. et al. Gene therapy with interleukin-10 receptor and interleukin-12 induces a protective interferon-γ-dependent response against B16F10-Nex2 melanoma. Cancer Gene Ther 18, 110–122 (2011). https://doi.org/10.1038/cgt.2010.58

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