重组病毒MVA-S的构建及其在小鼠中的免疫效果  

Construction of recombinant MVA-S and its immune effect in mice

作　　者：臧晚春 潘瑞[1] 陈国敏[1] 毕胜利[1] 曾毅[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室,北京102206

出　　处：《中华实验和临床病毒学杂志》2016年第2期149-152,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的 构建表达HBsAg重组MVA病毒,鉴定其抗原性并探讨不同免疫策略对免疫效果的影响.方法 通过基因克隆的方法构建携带目的基因的重组pSC11质粒,该重组质粒与MVA病毒共转染BHK-21细胞后,发生同源重组,经数轮蓝白斑筛选后,获得携带目的基因的重组病毒MVA-S.通过PCR和Western Blot鉴定目的基因的表达.分别采用单独免疫和联合免疫的策略免疫BALB/c小鼠,通过ELISpot检测细胞免疫水平.结果 PCR和Western Blot证实MVA-S携带目的基因并具有抗原性.ELISpot结果显示MVA-S能诱导小鼠产生特异性细胞免疫,MVA-S与DNA-S疫苗单独免疫效果无统计学差异;DNA-S疫苗和MVA-S联合免疫效果强于DNA-S疫苗单独免疫;等剂量DNA-S疫苗初始免疫,高剂量MVA-S加强免疫效果更强;2种DNA疫苗（pVR-S、pcDNA-S）分别初始免疫、等剂量MVA-S加强免疫,组间差异无统计学意义.结论 研究成功构建了表达目的基因的重组病毒MVA-S,该重组病毒可诱导小鼠产生特异性细胞免疫,联合免疫效果更强.Objective To construct the recombinant modified vaccinia Ankara vaccine expressing HBsAg and investigate its antigenicity and immune effect by different strategies.Methods Target gene was cloned into plasmid pSC1l,the recombinant plasmid pSC11-S was transfected into BHK-21 cells that infected with MVA.Homologous recombination occurred between MVA and pSC1 1-S.The recombinant virus MVA-S was selected by several rounds of blue/clear plaques.Target gene was identified by PCR.The expression of target gene was analyzed by Western Blot.The BALB/c mice were immunized with single immunization and consecutive immunization strategy.The level of specific cellular response was measured by ELISpot assays.Results The recombinant virus expressing target gene was confirmed by PCR and Western Blot showed that MVA-S could elicit specific cellular response.There were no significant differences among two kinds of DNA vaccine and recombinant MVA-S.The prime/boost regimen could induce higher cellular response than single DNA vaccine immunization.High doses of MVA-S boost with the same DNA vaccine prime could induce highest response.There was no significant difference between different DNA vaccines （pVR-S,pcDNA-S） followed by same MVA-S boost.Conclusions MVA-S expressing target was constructed successfully,and it could induce specific cellular immune response which was higher with the strategy of DNA prime/MVA-S boost than that of single immunization.

关 键 词：痘苗病毒安卡拉株 乙肝表面抗原 联合免疫 

分 类 号：R392[医药卫生—免疫学]