Anti-Tumor Effects after Adoptive Transfer of IL-12 Transposon-Modified Murine Splenocytes in the OT-I-Melanoma Mouse Model

• Published in: PloS one Vol. 10; no. 10; p. e0140744
• Main Authors: Galvan, Daniel L, O'Neil, Richard T, Foster, Aaron E, Huye, Leslie, Bear, Adham, Rooney, Cliona M, Wilson, Matthew H
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 16-10-2015; Public Library of Science (PLoS)
• Subjects: Adoptive immunotherapy; Adoptive Transfer; Animal models; Animals; Anticancer properties; Antigens; Cancer; Cancer therapies; Cytokines; Deoxyribonucleic acid; DNA; DNA Transposable Elements; Fibroblasts; Gene therapy; HeLa Cells; Homing; House mouse; Humans; Immune response; Immune system; Immunodeficiency; Immunotherapy; Interleukin 12; Interleukin-12 - biosynthesis; Interleukin-12 - genetics; Interleukins; Life assessment; Luciferase; Lymphocytes; Lymphocytes T; Medicine; Melanoma; Melanoma - genetics; Melanoma - pathology; Melanoma - therapy; Mice; Mice, Inbred NOD; Mice, SCID; Neoplasms, Experimental - genetics; Neoplasms, Experimental - pathology; Neoplasms, Experimental - therapy; Ovalbumin; Pediatrics; Plasmids; Spleen; Splenocytes; Surgical implants; T cell receptors; T cells; T-Lymphocytes - metabolism; T-Lymphocytes - transplantation; Transposons; Tumors; Vectors (Biology); Xenografts; Xenotransplantation; United States > US; Tennessee; Texas; Nashville Tennessee
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0140744

Adoptive transfer of gene modified T cells provides possible immunotherapy for patients with cancers refractory to other treatments. We have previously used the non-viral piggyBac transposon system to gene modify human T cells for potential immunotherapy. However, these previous studies utilized adoptive transfer of modified human T cells to target cancer xenografts in highly immunodeficient (NOD-SCID) mice that do not recapitulate an intact immune system. Currently, only viral vectors have shown efficacy in permanently gene-modifying mouse T cells for immunotherapy applications. Therefore, we sought to determine if piggyBac could effectively gene modify mouse T cells to target cancer cells in a mouse cancer model. We first demonstrated that we could gene modify cells to express murine interleukin-12 (p35/p40 mIL-12), a transgene with proven efficacy in melanoma immunotherapy. The OT-I melanoma mouse model provides a well-established T cell mediated immune response to ovalbumin (OVA) positive B16 melanoma cells. B16/OVA melanoma cells were implanted in wild type C57Bl6 mice. Mouse splenocytes were isolated from C57Bl6 OT-I mice and were gene modified using piggyBac to express luciferase. Adoptive transfer of luciferase-modified OT-I splenocytes demonstrated homing to B16/OVA melanoma tumors in vivo. We next gene-modified OT-I cells to express mIL-12. Adoptive transfer of mIL-12-modified mouse OT-I splenocytes delayed B16/OVA melanoma tumor growth in vivo compared to control OT-I splenocytes and improved mouse survival. Our results demonstrate that the piggyBac transposon system can be used to gene modify splenocytes and mouse T cells for evaluating adoptive immunotherapy strategies in immunocompetent mouse tumor models that may more directly mimic immunotherapy applications in humans.