Gene Therapy against Murine Melanoma B16F10-Nex2 Using IL-13Rα2-Fc Chimera and Interleukin 12 in Association with a Cyclopalladated Drug

摘要

Interleukin 13 (IL-13) is immunoregulatory in many diseases, including cancer. The protective or suppressive role of CD1-restricted natural killer T cells (NKT cells) in tumor immunosurveillance and immunity is well documented. Interleukin 12 (IL-12) can activate type I NKT cells to produce interferon-gamma (IFN-γ), whereas type II NKT cells may produce IL-13. The high-affinity chain of IL-13Rα2 may act as negative inhibitor, suppressing the action of IL-13 and helping to maintain tumor immunosurveillance. We constructed an mIL-13Rα2-Fc chimera in a eukaryotic expression vector and confirmed the identity of the recombinant protein by immunoblot analysis and binding to IL-13 in chemiluminescent ELISA. Such DNA vaccine was tested against syngeneic B16F10-Nex2 murine melanoma. In vivo experiments showed a protective effect mediated by high production of IFN-γ and down-regulation of anti-inflammatory interleukins mainly by NKT 1.1⁺ T cells. Biochemoterapy in vivo with plasmid encoding mIL-13Rα2-Fc in association with plasmid encoding IL-12 and the 7A cyclopalladated drug led to a significant reduction in the tumor evolution with 30% tumor-free mice. We conclude that IL-12 gene therapy, followed by continuous administration of IL-13Rα2-Fc gene along with 7A-drug has antitumor activity involving the high production of proinflammatory cytokines and low immune suppression, specifically by NK1.1⁺T cells producing IL-13 and IL-10.