Expression of human clotting factor IX mediated by recombinant lentiviral vector in cultured cells and hemophilia B mice

摘要

To explore the expression of human clotting factor IX (hFIX) cDNA in vitro and the feasibility of gene therapy for hemophilia B mice mediated by recombinant lentiviral vector, a recombinant hFIX lentiviral vector driven by ubiquitin-C promoter, FUXW, and by ABP liver specific promoter, FAXW, was constructed respectively. Recombinant lentivirus was harvested from 293T cells by calcium phosphate-mediated transient cotransfection of three plasmids (transgene vector, CMV DELTA R8.2, VSV-G). hFIX expression was detected in supernatant of 293T, BHK and L-02 cells infected with FUXW virus, whereas higher expression of hFIX levels (630 ng/10~6 cells/48 h) was detected only in L-02 cells infected with FAXW virus. Serum hFIX antigen was detected in all hemophilia B mice treated with FAXW virus by tail vein injection, an efficiency level of hFIX was observed (45 ng/mL, approximately 1 percent of normal human levels), the expression lasted for more than 60 d. The results indicated that HIV-based lentiviral vectors offer a promising approach to the gene therapy of hemophilia B.