A system for functional analysis of Ebola virus glycoprotein
A system for functional analysis of Ebola virus glycoprotein
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DOI：
10.1073/pnas.94.26.14764
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发表时间：
1997-12-23
影响因子：
11.1
通讯作者：
Kawaoka, Y
中科院分区：
文献类型：
--
作者：
Takada, A;Robison, C;Kawaoka, Y
埃博拉病毒在人类和非人类灵长类动物中引起出血热，导致高达90%的死亡率。对这种病毒的研究因其特殊的致病性而受阻，需要生物安全4级控制。为了解决这个问题，我们开发了一种新的互补系统用于埃博拉病毒糖蛋白的功能分析，它依赖于含有绿色荧光蛋白基因而不是受体结合G蛋白基因（VSV Delta G*）的重组水泡性口炎病毒（VSV）。在此，我们表明，埃博拉病毒莱斯顿病毒糖蛋白（ResGP）有效地纳入VSV颗粒。这种具有整合的ResGP的重组VSV（VSV Delta G*-ResGP）以与埃博拉病毒的宿主范围嗜性一致的方式比所检查的任何其他哺乳动物或禽类细胞更有效地感染灵长类细胞，而补充有VSV G蛋白的VSV Delta G*（VSV Delta G*-G）有效地感染大多数测试的细胞。我们还测试了该系统用于研究埃博拉病毒细胞受体的实用性。化学修饰细胞以改变其表面蛋白质显著降低了其对VSV Delta G*-ResGP的易感性，但对VSV Delta G*-G的易感性没有降低。这些发现表明，具有N-连接寡糖链的细胞表面糖蛋白有助于埃博拉病毒的进入，可能作为病毒进入的特异性受体和/或辅因子。因此，我们的VSV系统应该是有用的研究功能的糖蛋白从高致病性病毒或那些不能在体外培养。
Ebola virus causes hemorrhagic fever in humans and nonhuman primates, resulting in mortality rates of up to 90%. Studies of this virus have been hampered by its extraordinary pathogenicity, which requires biosafety level 4 containment. To circumvent this problem, we developed a novel complementation system for functional analysis of Ebola virus glycoproteins, It relies on a recombinant vesicular stomatitis virus (VSV) that contains the green fluorescent protein gene instead of the receptor-binding G protein gene (VSV Delta G*). Herein we show that Ebola Reston virus glycoprotein (ResGP) is efficiently incorporated into VSV particles. This recombinant VSV with integrated ResGP (VSV Delta G*-ResGP) infected primate cells more efficiently than any of the other mammalian or avian cells examined, in a manner consistent with the host range tropism of Ebola virus, whereas VSV Delta G* complemented with VSV G protein (VSV Delta G*-G) efficiently infected the majority of the cells tested. We also tested the utility of this system for investigating the cellular receptors for Ebola virus. Chemical modification of cells to alter their surface proteins markedly reduced their susceptibility to VSV Delta G*-ResGP but not to VSV Delta G*-G. These findings suggest that cell surface glycoproteins with N-linked oligosaccharide chains contribute to the entry of Ebola viruses, presumably acting as a specific receptor and/or cofactor for virus entry. Thus, our VSV system should be useful for investigating the functions of glycoproteins from highly pathogenic viruses or those incapable of being cultured in vitro.