Constructon Of The MIL-12Producing Recombinant Lentiviral Vector And Its Transfection Effect On Cells Contained In Skin Flap

Interleukin-12is a potent cytokine with a critical role in the regulation of cellular immune response and has been demonstrated to both inhibit establishment of tumors and induce regression of established tumors. However, the use of IL-12for immuneotherapy has been limited by the toxicity and shot expression associated with both systemic and regional in vivo administration. Microvascular skin flap administered by arterial delivery is an ideal plastic surgical delivery system for targeted gene therapy because of the abandent microvascular bed, highly efficient transduction of cells within the flap, minimal systemic transduction of uninvolved tissues of organs, minimizing systemic toxicity, extremely high titers of vectors that can be safely administered, and reversible procedure which can prevent the adverse reaction to the viral vector or the therapeutic transgene. Lentiviral vector is believed to be most perfect vector with improved efficiency, specificity, and safety. We suggested an innovative method to solve these problems:gene therapy by administering LV-mIL-12to the microvascular flap through arterial delivery. The methods of the present study are showed as below. First, we construct and identify LV-mIL-12. Second, we compare the transduct efficiencies of lentivector and adenovector infecting dermal fibrosis and vascular endothelial cell with different enhance infect materials. At last, we test the relationship between infection time and transduction efficiency. The results show that LV-mIL-12we constructed can infect cells in skin flap ex vivo efficiently, and secret mIL-12with biological function. Moreover, the transduction efficiency of lentivector is high when adding polybrene or Enhance infection solution. In addition, the transduction efficiency is associated with the transduction time.