Establishment of a Nipah virus rescue system
Establishment of a Nipah virus rescue system
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DOI：
10.1073/pnas.0606972103
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发表时间：
2006-10
期刊：
影响因子：
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通讯作者：
M. Yoneda;V. Guillaume;Fusako Ikeda;Yuki Sakuma;Hiroki Sato-;T. Wild;C. Kai
中科院分区：
文献类型：
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作者：
M. Yoneda;V. Guillaume;Fusako Ikeda;Yuki Sakuma;Hiroki Sato-;T. Wild;C. Kai
尼帕病毒（NiV）是一种副粘病毒，于1998年在马来西亚的一次猪和人类感染暴发中首次发现，并在人类中引起高死亡率。果蝠生活在从印度到西太平洋的广大地区，被确定为病毒的天然宿主。然而，导致人类严重致病性（高达70%的死亡率）和能够跨越物种屏障的机制尚不清楚。在这项研究中，我们建立了一个系统，使救援复制NiVs从克隆的DNA共转染构建的全长cDNA克隆和支持质粒编码病毒核蛋白，磷蛋白，和聚合酶的重组牛痘病毒，MVAGKT 7，表达T7 RNA聚合酶的感染。通过使用新开发的反向遗传学系统，拯救的NiV（rNiV）在体外表现出与亲本病毒相似的特性，并在先前通过实验感染建立的动物模型中保留了严重的致病性。还开发了表达增强型绿色荧光蛋白（rNiV-EGFP）的重组NiV。使用该病毒，在许多不同来源的细胞系中将NiV的允许性与已知细胞受体肝配蛋白B2的存在进行了比较。有趣的是，表达肝配蛋白B2的两个细胞系对rNiV-EGFP不敏感，表明完全NiV复制显然需要额外的因子。NiV的反向遗传学研究将为分析NiV致病性和跨种感染的分子机制提供有力的工具。
Nipah virus (NiV), a paramyxovirus, was first discovered in Malaysia in 1998 in an outbreak of infection in pigs and humans and incurred a high fatality rate in humans. Fruit bats, living in vast areas extending from India to the western Pacific, were identified as the natural reservoir of the virus. However, the mechanisms that resulted in severe pathogenicity in humans (up to 70% mortality) and that enabled crossing the species barrier were not known. In this study, we established a system that enabled the rescue of replicating NiVs from a cloned DNA by cotransfection of a constructed full-length cDNA clone and supporting plasmids coding virus nucleoprotein, phosphoprotein, and polymerase with the infection of the recombinant vaccinia virus, MVAGKT7, expressing T7 RNA polymerase. The rescued NiV (rNiV), by using the newly developed reverse genetics system, showed properties in vitro that were similar to the parent virus and retained the severe pathogenicity in a previously established animal model by experimental infection. A recombinant NiV was also developed, expressing enhanced green fluorescent protein (rNiV-EGFP). Using the virus, permissibility of NiV was compared with the presence of a known cellular receptor, ephrin B2, in a number of cell lines of different origins. Interestingly, two cell lines expressing ephrin B2 were not susceptible for rNiV-EGFP, indicating that additional factors are clearly required for full NiV replication. The reverse genetics for NiV will provide a powerful tool for the analysis of the molecular mechanisms of pathogenicity and cross-species infection.