Characterization of a novel bat-HKU2-like swine enteric alphacoronavirus (SeACoV) infection in cultured cells and development of a SeACoV infectious clone
Characterization of a novel bat-HKU2-like swine enteric alphacoronavirus (SeACoV) infection in cultured cells and development of a SeACoV infectious clone
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培养细胞中新型蝙蝠-HKU2 样猪肠道 α 冠状病毒 (SeACoV) 感染的表征以及 SeACoV 感染性克隆的开发
DOI：
10.1016/j.virol.2019.08.006
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发表时间：
2019-10-01
期刊：
影响因子：
3.7
通讯作者：
Huang, Yao-Wei
中科院分区：
文献类型：
--
作者：
Yang, Yong-Le;Liang, Qi-Zhang;Huang, Yao-Wei
猪肠道甲型冠状病毒（SeACoV），又称猪急性腹泻综合征冠状病毒（SADSCoV），属于菊头蝠冠状病毒HKU2种。在此，我们报告SeACoV的体外初步特性。我们制备了四种针对SeACoV刺突蛋白、膜蛋白、核衣壳蛋白及非结构蛋白3的抗体，这些抗体能够与SeACoV感染的Vero细胞中的病毒抗原发生反应。我们基于细胞适应第10代病毒构建了DNA启动的SeACoV感染性克隆，并在培养细胞中拯救出带有独特遗传标记的重组病毒。在SeACoV感染的细胞中，通过实验鉴定出6种包含前导-主体连接位点的亚基因组mRNA，其中一种双顺反子mRNA编码辅助蛋白NS7a和NS7b基因。通过电子显微镜观察到SeACoV感染诱导的细胞超微结构变化。SeACoV感染性克隆以及针对不同病毒蛋白的一系列抗体的获得，将有助于进一步研究，以了解SeACoV复制和致病的分子机制。
Swine enteric alphacoronavirus (SeACoV), also known as swine acute diarrhea syndrome coronavirus (SADSCoV), belongs to the species Rhinolophus bat coronavirus HKU2. Herein, we report on the primary characterization of SeACoV in vitro. Four antibodies against the SeACoV spike, membrane, nucleocapsid and nonstructural protein 3 capable of reacting with viral antigens in SeACoV-infected Vero cells were generated. We established a DNA-launched SeACoV infectious clone based on the cell adapted passage-10 virus and rescued the recombinant virus with a unique genetic marker in cultured cells. Six subgenomic mRNAs containing the leader-body junction sites, including a bicistronic mRNA encoding the accessory NS7a and NS7b genes, were experimentally identified in SeACoV-infected cells. Cellular ultrastructural changes induced by SeACoV infection were visualized by electron microscopy. The availability of the SeACoV infectious clone and a panel of antibodies against different viral proteins will facilitate further studies on understanding the molecular mechanisms of SeACoV replication and pathogenesis.