埃博拉和马尔堡病毒囊膜糖蛋白嵌合型重组水泡性口炎病毒的构建及鉴定  

Construction of chimeric vesicular stomatitis virus expressing glycoproteins of Ebola virus and Marburg virus

作　　者：邵钰[1] 王金良[1] 刘任强[1] 张会雷 王喜军[1] 葛金英[1] 温志远[1] 步志高[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2016年第6期438-442,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金："十二五"国家支撑计划课题(2013BAD12B05)

摘　　要：埃博拉病毒(EBOV)和马尔堡病毒(MARV)属于丝状病毒科,能够导致人和动物发生致命的埃博拉出血热和马尔堡出血热,这两种病是重要的尚未传入我国的烈性人兽共患病。为研制安全有效的储备疫苗,本研究利用水泡性口炎病毒(VSV)反向遗传操作技术,将VSV自身糖蛋白(GP)分别替换为EBOV或MARV的GP,构建了表达扎伊尔型EBOV、苏丹型EBOV及MARV GP的嵌合VSV重组病毒r VSV△G/ZEBOVGP、r VSV△G/SEBOVGP及r VSV△G/MARVGP。Western blot和间接免疫荧光试验表明这3种GP蛋白在重组病毒感染的细胞中均能够正确表达。电镜观察结果显示,3种重组病毒均与亲本病毒具有相同形态特征,表明这3种外源GP蛋白均能够嵌合在重组病毒粒子表面。本研究构建的这3种重组病毒为进一步的动物免疫试验并评价其免疫效果奠定了基础。Ebola virus and Marburg virus belong to the Filovirus family, which cause severe hemorrhagic fever with high mortality in humans and nonhuman primates. Ebola and Marburg hemorrhagic fever are very important zoonosis. To develop safe, effective candidate recombinant viral vaccines, three recombinant vesicular stomatitis viruses （rVSVAG/ZEBOVGP, rVSVAG/ SEBOVGP and rVSV AG/MARVGP） were generated based on VSV reverse genetics, which expressed a single filovirus glycoprotein （GP） in replace of the VSV GP. The results of western blot and immunofluorescence assay indicated that the three GPs in recombinant virus were expressed effectively in infected cells. The three recombinant viruses had the same morphological characteristics as the parental virus by transmission electron microscope, which suggested that the GPs were able to express on the surface of the recombinant virus particles. The results of this study indicated that the three recombinant viruses were rescued correctly and lay the foundation of fitrther study on animal immune experiment and evaluation of the immtme effect.

关 键 词：埃博拉病毒 马尔堡病毒 囊膜糖蛋白 水泡性口炎病毒 

分 类 号：S852.65[农业科学—基础兽医学]