汉滩病毒包膜糖蛋白假病毒的构建及特性研究

Construction and identification of hantaan pseudovirions

目的：研究汉滩病毒包膜糖蛋白与宿主细胞作用的分子机制,构建含有汉滩病毒（HTNV）标准株76-118株包膜糖蛋白的假病毒,并对其进行检测.方法：将表达汉滩病毒包膜糖蛋白的质粒和以VSVΔG为骨架的假病毒颗粒,与293T细胞共转染,构建汉滩病毒的假病毒,用中和实验,受体抑制实验,免疫共沉淀和Western Blot对其抗原性、中和反应、受体作用等进行检测.结果：汉滩病毒假病毒与活病毒有相似的抗原特性,汉滩病毒假病毒可以模拟活病毒进入细胞的方式,其糖蛋白是以二聚体形式存在的.结论：成功构建含有汉滩病毒标准株包膜糖蛋白的假病毒,它与活病毒的抗原特性和受体作用的方式相似,这为进一步研究汉滩病毒包膜糖蛋白与宿主细胞作用的分子机制提供了很好的工具.

AIM： To study the molecular mechanism of hantaanvirus（ HTNV） envelope glycoprotein with host cell,and constructe pseudovirions of HTNV 76- 118 strains. METHODS： It was constructed by incorporating of HTNV glycoproteins onto replication-defective vesicular stomatitis virus（ VSV） cores in which the gene for the surface G protein has been replaced by the reporter gene. Antigenicity,neutral reaction and receptor function were detected by neutralization test,co-immunoprecipitation and flow cytometry. RESULTS： There was good concordance of the antigenicity and the way inside host cells between HTNV and its pseudovirions. Also,the glycoprotein G N and G C of pseudovirions existed as heterodimers. CONCLUSION： The pseudovirions are successfully constructed and the pseudovirions can be used to simulate the way of HTNV entry and antigenicity. This study provides a good stool for study on molecular mechanism of hantaan virus envelope glycoprotein.