水泡性口炎病毒保护性抗原基因核酸疫苗质粒的构建及其免疫原性  

Construction and Immunogenictiy of Recombinant DNA Vaccine Plasmid with Protective Antigen Gene of Vesicular Stomatitis Virus

作　　者：周学章[1] 李元刚[1] 高丰[2] 

机构地区：[1]宁夏大学教育部重点实验室,银川750021 [2]吉林大学畜牧兽医学院,长春130062

出　　处：《中国生物制品学杂志》2008年第6期500-503,509,共5页Chinese Journal of Biologicals

基　　金：国家自然基金资助项目(30270994)

摘　　要：目的构建水泡性口炎病毒(VSV)保护性抗原基因的重组核酸疫苗质粒,并检测其免疫原性。方法利用RT-PCR从VSV中扩增G蛋白基因,构建重组表达质粒pIRES-G1500,转染BHK-21细胞,以间接免疫荧光、PCR、SDS-PAGE、ELISA和Westernblot检测质粒的表达,用核酸疫苗质粒免疫BALB/c小鼠,检测小鼠的细胞免疫和体液免疫水平。结果构建的重组核酸疫苗质粒在BHK-21细胞中获得正确表达,质粒免疫小鼠后,可刺激脾细胞、T淋巴细胞亚类数量及CTL杀伤活性显著升高,并可产生高滴度的抗VSV特异性抗体。结论该重组核酸疫苗质粒可作为VSV的候选疫苗。Objective To construct a recombinant DNA vaccine plasmid with protective antigen gene of vesicular stomatitis virus（VSV） and determine its immunogenicity. Methods Amplify the gene encoding G protein from the genome of VSV by RT-PCR to construct recombinant plasmid plRES-G1500. Transfect BHK-21 cells with plRES-G1500 and identify the expressed product by indirect IFA, PCR, SDS-PAGE, ELISA and Western blot. Immunize BALB / c mice with pIRES-G1500 and determine the cellular and humoral immune responses. Results The constructed recombinant DNA vaccine plasmid plRES-G1500 was correctly expressed in BHK-21 cells. The plasmid stimulated the significant increases of the numbers of splenic cells and T lymphocyte subclasses as well as the killing activity of CTL, and induced high specific antibody titer against VSV. Conclusion The constructed recombinant DNA plasmid might be used as a candidate vaccine against VSV.

关 键 词：水泡性口炎病毒 G蛋白 核酸疫苗 免疫原性 

分 类 号：Q786[生物学—分子生物学] R392-33[医药卫生—免疫学]