HCV多表位基因重组BCG的筛选及对其诱导的免疫应答研究  

Screening of the recombinant BCG expressing HCV multiepitope antigen and analysis of the immune re-spouses elicited in BALB/c mice

作　　者：韦三华[1] 尹文[2] 雷迎峰[2] 胡兴斌[2] 董轲[1] 李斌[1] 张青[1] 张利军[1] 张惠中[1] 

机构地区：[1]第四军医大学唐都医院临床实验与检验输血科,西安710038 [2]第四军医大学基础部微生物学教研室

出　　处：《中华微生物学和免疫学杂志》2007年第11期1041-1045,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金项目（30671878）;陕西省自然科学基金项目（2005C214）

摘　　要：目的将丙型肝炎病毒（HCV）多表位基因的穿梭质粒pDE22-CtEm电转化BCG，筛选重组BCG（rBCG），免疫BALB／c小鼠，研究其诱导的免疫应答。方法培养并制备BCG感受态，将重组穿梭质粒pDE22-CtEm电转化BCG，筛选rBCG。rBCG免疫小鼠，同时进行基因免疫，测定血清中特异性抗体，分离小鼠脾淋巴细胞，体外测定淋巴细胞刺激指数、IFN-γ和进行CTL杀伤实验，观察rBCG在小鼠体内诱导的体液和细胞免疫应答。结果筛选出HCV多表位基因rBCG，免疫小鼠后在小鼠体内诱导出特异性的体液和细胞免疫应答，免疫应答水平均高于重组的基因免疫。结论构建的HCV多表位抗原rBCG活疫苗优于基因疫苗。Objective To bolt recombinant BCG （rBCG） expressing HCV multiepitope gene after elec-trotransformation with reeombinant secreting shuttle expression vectors pDE22-CtEm, and analyze the immtme re-sponses in BALA/c miee. Methods Recombinant shuttle expression vector pDE22-CtEm was eleetrotransformed into eompetenee BCG, and the rBCG was bolted. BALA/c miee were immunized with rBCG, with the DNA immunization as the eontrol. The HCV speeifie antibodies were tested. After the spleen lymphoeytes were isolated, the lymph-proliferation response as well as the IFN-γ was measured, and the cytotoxic assay was earried out. Results The reeombinant BCG （rBCG） expressing HCV muhiepitope gene was bolted. The HCV speeifie humoral and eellular immune responses were eheited in BALB/e miee, which were higher than that of the DNA immunization. Conclusion The rBCG vaccine expressing HCV multiepitope gene has an advantage over DNA vaccine.

关 键 词：丙型肝炎病毒 表位 重组BCG 疫苗 

分 类 号：R686[医药卫生—骨科学]