腸道病毒71型(EV71)是引起手足口病的主要病原體,。手足口病是常發(fā)于5歲以下兒童的傳染性疾病,，部分患者會出現(xiàn)腦炎,、肺水腫以及癱瘓等嚴(yán)重神經(jīng)癥狀,，甚至死亡,，嚴(yán)重威脅我國兒童的健康,，但目前還沒有商品化的EV71疫苗來有效防治該疾病,。
EV71是單鏈正義RNA病毒，其病毒顆粒為球形,，具有一個對稱二十面體狀的蛋白質(zhì)外殼,，稱為衣殼。病毒衣殼主要起到保護(hù)病毒RNA,，協(xié)助病毒感染的作用,。同時，病毒衣殼通常具有良好的抗原性,，能夠有效誘發(fā)機(jī)體的體液免疫與細(xì)胞免疫應(yīng)答,，起到免疫防御作用，是疫苗研究的主要靶標(biāo)之一,。因此,，深入開展EV71病毒衣殼蛋白研究，將為盡快開發(fā)出安全有效的EV71疫苗提供重要信息和幫助,。
上海巴斯德研究所疫苗學(xué)與抗病毒策略課題組博士研究生劉慶偉等在黃忠研究員的指導(dǎo)下,，制備出針對EV71衣殼亞基蛋白的一系列多克隆抗體，并利用這些特異性抗體研究了EV71病毒顆粒衣殼蛋白組成情況以及衣殼的組裝過程,。結(jié)果顯示,，純化的無感染性病毒顆粒衣殼由VP0、VP1和VP3三個亞基蛋白共同組裝而成,，而感染細(xì)胞裂解液中具有感染性的病毒顆粒衣殼蛋白除了VP0,、VP1和VP3外，還檢測到了VP0的切割產(chǎn)物VP2,，提示VP0被切割為VP2和VP4與病毒的感染能力有重要相關(guān)性,。利用三種亞基蛋白特異性抗體進(jìn)行免疫熒光實(shí)驗(yàn)的結(jié)果顯示，病毒衣殼亞基蛋白共定位于被感染細(xì)胞的細(xì)胞質(zhì)中,，說明病毒衣殼在感染細(xì)胞胞質(zhì)中組裝,。
該項(xiàng)研究不僅加深了對EV71衣殼蛋白構(gòu)成及組裝的認(rèn)識，而且為EV71的臨床診斷、新型疫苗開發(fā)和質(zhì)量控制,、以及分子病毒學(xué)研究提供了新的研究工具和手段,。研究成果于10月6日在國際學(xué)術(shù)刊物 Journal of Virological Methods 在線發(fā)表。研究工作得到中國科學(xué)院“百人計(jì)劃”的經(jīng)費(fèi)支持,。（生物谷Bioon.com）
doi: 10.1016/j.jviromet.2012.09.024
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Characterization of enterovirus 71 capsids using subunit protein-specific polyclonal antibodies
Liu Q, Huang X, Ku Z, Wang T, Liu F, Cai Y, Li D, Leng Q, Huang Z.
Enterovirus 71 (EV71), a member of the Enterovirus genus of the Picornaviridae family, is one of the major causative agents of hand-foot-and-mouth disease (HFMD), which is prevalent in the Asia-Pacific region. In this article, a set of capsid subunit protein-specific antibodies was used to characterize the EV71 structural protein processing and to determine the composition and assembly of EV71 capsids. SDS-PAGE and Western blot analyses showed that the capsids of a purified EV71 preparation, which lacked viral infectivity, were composed of processed VP0, VP1 and VP3, all of which co-assembled into particles. Analyses of infectious EV71-containing cell lysate revealed the presence of VP2, in addition to VP0, VP1 and VP3, suggesting that the cleavage of VP0 into VP2 and VP4 is important for infectivity. Immunofluorescent staining with the three specific antibodies demonstrated that the capsid subunit proteins co-localized in the cytoplasm of cells infected with EV71. The results add new information on the processing, assembly and localization of EV71 capsid proteins, and demonstrate the usefulness of the capsid protein-specific antibodies for virological investigation and for development of vaccines and diagnostic reagents
