丙型肝炎病毒多表位基因核酸疫苗的构建及其免疫原性  

Construction and Immunogenicity of Recombinant Plasmid Carrying Multiepitope Gene of Hepatitis C Virus

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,西安710032

出　　处：《中国生物制品学杂志》2007年第9期633-636,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金项目(NO:30671878)

摘　　要：目的 构建丙型肝炎病毒（HCV）多表位基因的真核表达载体pcDNA3.1（-）-CtEm,用该重组质粒免疫BALB/c小鼠,并检测其免疫原性.方法 用BamHⅠ和Hind Ⅲ双酶切含有HCV C区、E2区模拟表位和NS3～NS5区细胞表位串联基因的原核表达载体pQE30-CtEm,克隆入真核表达载体pcDNA3.1（-）,脂质体瞬时转染CHO细胞,并检测其表达.以100 μg重组质粒免疫BALB/c小鼠,检测其体液免疫和细胞免疫效果.结果 所构建的真核表达载体pcDNA3.1（-）-CtEm在CHO细胞中能获得有效表达.该质粒免疫小鼠后可诱导高水平的抗体,特异性淋巴细胞增殖反应、IFN-γ水平及CTL活性均明显高于空载体和生理盐水对照组.结论 已成功构建HCV多表位基因的真核表达载体pcDNA3.1（-）-CtEm,免疫小鼠后可诱导特异性体液免疫和细胞免疫应答.Objective To construct a eukaryotic expression vector of hepatitis C virus （HCV） muhiepitope gene and study its immunogencity. Methods Digest prokaryotic expression vector pQE30-CtEm carrying the mimic epitopes at C and E2 domains and the tandem epitope at NS3-NS5 domain of HCV with BamH Ⅰ and Hind Ⅲ, and insert the obtained HCV multiepitope gene CtEm into eukaryotic expression vector pcDNA3.1 （-）. Transiently transfect CHO cells with the constructed recombinant plasmid in mediation of liposome and determine the expression of HCV multiepitope antigen. Immunize BALB/c mice with 100 μg of the constructed recombinant plasmid and determine the humoral and cellular immune responses. Results The constructed recombinant plasmid pcDNA3.1 （-）-CtEm was successfully expressed in CHO cells and induced high antibody levels in mice. The specific lymphocyte proliferation level, IFN-γ and CTL activity of mice immunized with the constructed recombinant plasmid were significantly higher than those with empty vector or physiological saline. Conclusion The eukaryotic expression vector pcDNA3. 1 （-） -CtEm of HCV multiepitope gene was successfully constructed and induced specific humoral and cellular immune responses in mice.

关 键 词：丙型肝炎病毒 多表位基因 核酸疫苗 免疫原性 

分 类 号：R373.21[医药卫生—病原生物学]