Ⅰ～Ⅳ型登革病毒包膜蛋白B细胞中和表位的鉴定  

Mapping of the B Cell Neutralizing Epitopes on EDⅢ of Envelope Protein from Dengue Virus

作　　者：林亚英 温坤[1] 郭勇晖[1] 丘立文[1] 潘玉先[1] 余楠[1] 狄飚[2] 陈月[1,3] 

机构地区：[1]南方医科大学珠江医院检验医学部,广州510282 [2]广州市疾病预防控制中心,广州510440 [3]广州军区疾病预防控制中心,广州510507

出　　处：《病毒学报》2015年第6期665-673,共9页Chinese Journal of Virology

基　　金：国家自然科学基金资助(81201296);广东省科技计划项目资助(2014A020212252;2015a020218003)

摘　　要：登革病毒包膜蛋白（Dengue virus envelope protein,DENV E）是诱导中和抗体主要的蛋白,登革病毒包膜蛋白Ⅲ区（Dengue virus envelope protein domainⅢ,DENV EDⅢ）是构建DENV亚单位疫苗的主要靶标,但是其B细胞中和表位目前了解不多。我们采用两组覆盖DENV-1EDⅢ的重叠多肽（12肽和16肽）同27株针对DENV-1EDⅢ中和单抗反应,筛选DENV EDⅢ上B细胞表位。采用该方法,我们发现了一高度保守的Ⅰ～Ⅳ型DENV共同交叉中和B细胞表位和一高度保守的DENV-1血清型特异性B细胞中和表位,分别位于DENV-1E蛋白氨基酸残基序列第309~320位和第381~392位（Amino acid residues 309~320,and 381~392;aa 309~320和381~392）。Ⅰ～Ⅳ型DENV共同交叉中和B细胞表位在Ⅰ～Ⅳ型DENV分离株中存在高度保守共同序列310 KEVAETQHGT319,DENV-1E蛋白中E309、V312、A313和V320不影响蛋白抗原性。DENV-1血清型特异性B细胞中和表位（DENV-1E蛋白氨aa 381~392）在DENV-1分离株中高度保守,在黄病毒属其它病毒中不保守。我们也发现一具有DENV-1分离株特异性的B细胞中和表位位于DENV-1E蛋白氨基酸残基序列第329~348位。这些新发现的DENV-1E蛋白EDⅢ上B细胞中和表位可能有助于研制新的DENV亚单位疫苗。Dengue virus（DENV）envelope[E]protein is the major surface protein of the virions that indued neutralizing antibodies.The domain III of envelope protein（EDIII）is an immunogenic region that holds potential for the development of vaccines;however,the epitopes of DENV EDIII,especially neutralizing B-cell linear epitopes,have not been comprehensively mapped.We mapped neutralizing B-cell linear epitopes on DENV-1EDIII using 27 monoclonal antibodies against DENV-1EDIII proteins from mice immunized with the DENV-1EDIII.Epitope recognition analysis was performed using two set of sequential overlapping peptides（16mand 12m）that spanned the entire EDIII protein from DENV-1,respectively.This strategy identified a DENV-1type-specific and a group-specific neutralizing epitope,which were highly conserved among isolates of DENV-1and the four DENV serotypes and located at two regions from DENV-1E,namely amino acid residues 309-320 and 381-392（aa 309-320 and 381-392）,respectively.aa310~319（310 KEVAETQHGT319）was similar among the four DENV serotypes and contact residues on aa 309~320from E protein were defined and found that substitution of residues E309,V312,A313 and V320 in DENV-2,-3,-4isolates were antigenically silent.We also identified a DENV-1type-specific strain-restricted neutralizing epitope,which was located at the region from DENV-1E,namely amino acid residues329-348.These novel type-and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.

关 键 词：登革病毒 包膜蛋白Ⅲ区 中和抗体 B细胞表位 

分 类 号：R373.9[医药卫生—病原生物学]