Characteristics of enterovirus 71-induced cell death and genome scanning to identify viral genes involved in virus-induced cell apoptosis
Characteristics of enterovirus 71-induced cell death and genome scanning to identify viral genes involved in virus-induced cell apoptosis
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肠道病毒71诱导细胞死亡的特征和基因组扫描以确定参与病毒诱导细胞凋亡的病毒基因
DOI：
10.1016/j.virusres.2019.03.017
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发表时间：
2019-05-01
期刊：
影响因子：
5
通讯作者：
Long, Jian-Er
中科院分区：
文献类型：
--
作者：
Bai, Jinjin;Chen, Xixi;Long, Jian-Er
肠道病毒71型（EV 71）可引起婴幼儿手足口病和严重的神经并发症。病毒的发病机制与病毒诱导的细胞死亡和炎性细胞因子的产生有关，这通常与程序性细胞死亡的类型有关。通过显微镜、细胞染色和免疫印迹分析EV 71感染的细胞，以确定EV 71诱导的细胞死亡的特征。结果表明，EV 71感染诱导细胞皱缩，核浓缩，线粒体电位降低，和膜磷脂酰丝氨酸易位。在病毒诱导的细胞死亡过程中，还观察到胱天蛋白酶-9活化、聚（ADP-核糖）聚合酶裂解和乳酸脱氢酶释放。未检测到活化的gasdermin D（GSDMD）和磷酸化的混合谱系激酶结构域样蛋白（p-MLKL）。这些观察结果表明，EV 71诱导的细胞死亡主要是通过细胞凋亡通过内在途径，而不是由GSDMD介导的焦亡和p-MLKL介导的坏死性凋亡。基因组扫描分析表明，EV 71 2A、2B和3C可能是病毒诱导细胞死亡的决定基因。进一步的实验显示，EV 71 2A-和3C-诱导的细胞死亡表现出对它们的蛋白酶活性的依赖性，但涉及不同的机制。EV 71 2A诱导的细胞死亡与宿主帽依赖性翻译的关闭相关，而EV 71 3C诱导的细胞死亡可能不归因于此机制。这些发现将加强我们对EV 71感染和病毒发病机制的理解，并有助于确定抗病毒靶点。
Enterovirus 71 (EV71) causes hand-foot-and-mouth disease and severe neural complications in infants and young children. Viral pathogenesis is associated with virus-induced cell death and inflammatory cytokine production, which is usually correlated with the type of programmed cell death. EV71-infected cells were analyzed through microscopy, cell staining, and immunoblotting to determine the characteristics of EV71-induced cell death. Results demonstrated that EV71 infection induced cell shrinkage, nuclear condensation, decreased mitochondrial potential, and membrane phosphatidylserine translocation. Caspase-9 activation, poly (ADP-ribose) polymerase cleavage, and lactate dehydrogenase release were also observed during virus-induced cell death. The activated gasdermin D (GSDMD) and the phosphorylated mixed lineage kinase domain-like protein (p-MLKL) were not detected. These observations indicated that EV71-induced cell death was mainly executed by apoptosis through the intrinsic pathway rather than by GSDMD-mediated pyroptosis and p-MLKL-mediated necroptosis. Genome scanning analysis identified that EV71 2A, 2B, and 3C might be the determinant genes of virus-induced cell death. Further experiments showed that EV71 2A- and 3C-induced cell death exhibited dependence on their protease activities but involved different mechanisms. EV71 2A-induced cell death was correlated with the shutoff of host cap-dependent translation, whereas EV71 3C-induced cell death might not be ascribed to this mechanism. These findings would enhance our understanding of EV71 infection and viral pathogenesis, and help identify antiviral targets.