Construction and identification of a recombinant adenovirus expressing bioactive GM-CSF: Implication for GM-CSF secreting tumor vaccines design

Abstract

IntroductionGranulocyte-monocyte colony stimulating factor (GM-CSF) has been reported to to mediate antitumor effects through stimulation of various antigen presentation cells. Clearly, these properties make GM-CSF a potent adjuvant. Therefore, an oncolytic adenovirus coding for GMCSF was engineered. ObjectiveContraction, identification and characterization of recombinant adenovirus encoding GM-CSF. MethodsIn this study, a recombinant adenovirus expressing GM-CSF (rAd-GM) was constructed and characterized. Homologous recombination in bacteria followed by transfection of recombinant plasmid into a mammalian packaging cell line, viral production was conveniently followed by the gene incorporated into the viral backbone. Results & DiscussionWe also showed that the construct produced a 12 kDa recombinant protein which stimulated colony formation in bone marrow cells in vitro, indicating that the recombinant GM-CSF was biologically active. ConclusionFuture work will demonstrate the rAd-GM mediated antitumor immunity in rats with experimentally induced breast cancer.