SARS-coronavirus-2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology.
SARS-coronavirus-2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology.
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DOI：
10.1099/jgv.0.001453
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发表时间：
2020-09
期刊：
影响因子：
--
通讯作者：
Snijder EJ
中科院分区：
文献类型：
--
作者：
Ogando NS;Dalebout TJ;Zevenhoven-Dobbe JC;Limpens RWAL;van der Meer Y;Caly L;Druce J;de Vries JJC;Kikkert M;Bárcena M;Sidorov I;Snijder EJ
2019 年底，中国湖北省突然出现严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 及其随后的大流行，凸显了了解冠状病毒感染和发病机制的完整分子细节的重要性。在这里，我们比较了 SARS-CoV-2 和 SARS-CoV 的多种复制特征，并分析了这两种密切相关的病毒在常用的 Vero E6 细胞系中引起的细胞病理学。与 SARS-CoV 相比，SARS-CoV-2 产生的细胞内病毒 RNA 水平更高，但从培养基中回收的传染性病毒后代却少了约 50 倍。对 SARS-CoV-2 感染细胞进行免疫荧光显微镜检查，发现先前针对 SARS-CoV 的各种非结构蛋白、膜和核衣壳蛋白产生的抗血清具有广泛的交叉反应性。电子显微镜显示，两种 SARS 病毒引起的超微结构变化非常相似，并且发生在感染后相当的时间内。此外，我们确定这两种病毒对三种已确定的冠状病毒复制抑制剂（瑞德西韦、阿拉泊韦和氯喹）的敏感性非常相似，但 SARS-CoV-2 感染对用聚乙二醇化干扰素 α 预处理的细胞更为敏感。这两种病毒之间的一个重要区别是，在 Vero E6 细胞中传代后，SARS-CoV-2 显然面临着强大的选择压力，需要在其刺突蛋白基因中获得适应性突变。这些突变改变或删除了连接 S1 和 S2 结构域的区域中假定的弗林蛋白酶样切割位点，并导致噬菌斑测定中非常显着的表型变化。
The sudden emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the end of 2019 from the Chinese province of Hubei and its subsequent pandemic spread highlight the importance of understanding the full molecular details of coronavirus infection and pathogenesis. Here, we compared a variety of replication features of SARS-CoV-2 and SARS-CoV and analysed the cytopathology caused by the two closely related viruses in the commonly used Vero E6 cell line. Compared to SARS-CoV, SARS-CoV-2 generated higher levels of intracellular viral RNA, but strikingly about 50-fold less infectious viral progeny was recovered from the culture medium. Immunofluorescence microscopy of SARS-CoV-2-infected cells established extensive cross-reactivity of antisera previously raised against a variety of non-structural proteins, membrane and nucleocapsid protein of SARS-CoV. Electron microscopy revealed that the ultrastructural changes induced by the two SARS viruses are very similar and occur within comparable time frames after infection. Furthermore, we determined that the sensitivity of the two viruses to three established inhibitors of coronavirus replication (remdesivir, alisporivir and chloroquine) is very similar, but that SARS-CoV-2 infection was substantially more sensitive to pre-treatment of cells with pegylated interferon alpha. An important difference between the two viruses is the fact that – upon passaging in Vero E6 cells – SARS-CoV-2 apparently is under strong selection pressure to acquire adaptive mutations in its spike protein gene. These mutations change or delete a putative furin-like cleavage site in the region connecting the S1 and S2 domains and result in a very prominent phenotypic change in plaque assays.
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影响因子：
13.2
作者：
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通讯作者：
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影响因子：
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影响因子：
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DOI：
10.1099/vir.0.052910-0
发表时间：
2013-08
期刊：
The Journal of general virology
影响因子：
--
作者：
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影响因子：
7.6
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