乙型脑炎病毒样颗粒疫苗的制备及其免疫保护效率的初步评价  

Generation of Japanese Encephalitis Virus-likeParticle Vaccine and Preliminary Evaluation of Its Protective Efficiency

作　　者：张艳芳[1,2] 杜瑞坤 黄少梅[2] 张涛[1] 刘金良[1] 朱碧波[2] 王华林[1] 邓菲[1] 曹胜波[2] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071 [2]华中农业大学动物医学院,武汉430070

出　　处：《病毒学报》2016年第2期150-155,共6页Chinese Journal of Virology

基　　金：农业公益性行业专项(201203082)

摘　　要：将乙型脑炎病毒prME及其信号肽基因片段克隆到穿梭载体AcBacmid中,构建重组穿梭载体AcBac-prME。Lipofectine介导其转染Sf9细胞,获得重组杆状病毒Ac-prME。Western-blotting、间接免疫荧光试验及电镜观察,证实Ac-prME介导prME在Sf9细胞中高效表达,且形成了病毒样颗粒。小鼠免疫和攻毒试验表明,该病毒样颗粒免疫可以使小鼠获得有效抵抗乙型脑炎病毒强毒株攻击的能力,保护率高达100%,从而为研制基于乙型脑炎病毒样颗粒的亚单位疫苗奠定了基础。The cDNA fragment of JEV prME gene was cloned into the baculovirus shuttle vector（bacmid）to construct a recombinant baculovirus vector,defined as AcBac-prME.Then the recombinant baculovirus Ac-prME was obtained by transfecting Sf9 cells with AcBac-prME.Western blot analysis and immunofluorescence results indicated that both prM and E proteins were efficiently expressed in Sf9 cells.Electron microscopy suggested that prME was assembled into JEV-VLPs.To further evaluate the potential of JEVVLPs as vaccine,the mice were immunized with JEV-VLPs and then challenged with lethal JEV.The results of mice survival and pathological changes demonstrated that the JEV-VLPs performed complete protection against JEV-P3 strain and relieved pathological changes in the mice brain significant.This study suggest that JEV-VLPs would be a potential vaccine for Japanese encephalitis virus.

关 键 词：乙型脑炎病毒 杆状病毒 prME蛋白 病毒样颗粒 亚单位疫苗 

分 类 号：S852.43[农业科学—基础兽医学]