单纯疱疹病毒Ⅱ型细胞毒性T淋巴细胞表位重组DNA疫苗的构建表达及鉴定  

The construction and identify of herpes simplex virus Ⅱ cytotoxic T lymphocyte epitoperecombinant DNA vaccine

作　　者：关蕾[1] 杨慧兰[1] 樊建勇[1] 

机构地区：[1]广州军区广州总医院皮肤科,广东广州510010

出　　处：《临床皮肤科杂志》2006年第6期351-353,共3页Journal of Clinical Dermatology

基　　金：国家自然科学基金资助项目(3037290)

摘　　要：目的:针对单纯疱疹病毒Ⅱ型(HSV-2)包膜糖蛋白gD和gB的细胞毒性T淋巴细胞(cytotoxicTlymphocyte,CTL)表位,构建重组真核表达质粒pVAX-gD-CTL。方法:用PCR方法从已构建好的pc-gD真核表达质粒中扩增gD片段,将其与CTL表位共同连接到pVAX1载体,构建重组真核表达质粒pVAX-gD-CTL。采用免疫组化和Western-blotting法鉴定重组质粒的表达情况,免疫BALB/c小鼠,进行CTL活性鉴定。结果:构建的重组质粒能在非洲绿猴肾细胞(COS-7细胞)内表达。该重组DNA疫苗免疫接种BALB/c小鼠后,其CTL活性增高。结论:成功构建HSV-2pVAX-gD-CTL重组真核表达质粒,并能在真核细胞内表达,该DNA疫苗对细胞免疫有明显的促进作用。Objective： To construct a recombinant eukaryotic plasmids pVAX-gD-CTL based on HSV-2 gB and gD protein coding-gene. Methods： HSV-2 gD gene was amplified from pc-gD by PCR. The gD protein coding-gene and gB protein coding-CTL epitope were inserted to pVAX1 plasmid. The recombinant plasmid pVAX-gD-CTL was used to inoculate mice by intramascular injection. The CTL activity was detected by lactic dehydrogenase （LDH） assay. Results： Recombinant plasraids were confirmed by restriction and DNA sequencing, and gD protein was detected in COS-7 cells. The CTL activity was increased. Conclusions： Recombinant eukaryotic plasmids have been successfully constructed. The cell-mediated immune re- sponse could be induced when BALB/c mice were immunized with pVAX-gD-CTL DNA vaccine.

关 键 词：疱疹病毒Ⅱ型 蛋白质gD T淋巴细胞 细胞毒性 疫苗 DNA 

分 类 号：R752.1[医药卫生—皮肤病学与性病学]