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

作     者：ZHUHuanzhang CHENXiaoguang LIFeng GONGJuli XUEJinglun 

作者机构：Theauthorsmadeequalcontributionstothisreport. StateKeyLaboratoryofGeneticEngineeringInstituteofGeneticsSchoolofLifeSciencesFudanUniversityShanghai200433China 

出 版 物：《Chinese Science Bulletin》 (CHINESE SCIENCE BULLETIN)

年 卷 期：2003年第48卷第20期

页      面：2196-2200页

核心收录：

学科分类：0710[理学-生物学] 1002[医学-临床医学] 07[理学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 071007[理学-遗传学] 10[医学] 

基　　金：国家高技术研究发展计划(863计划)(2001AA217181) 

主　　题：血友病B 大鼠 基因治疗 人凝血因子Ⅸ基因 细胞培养 

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腞8.2, 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/106 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% 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.