Enterovirus A71 Induces Neurological Diseases and Dynamic Variants in Oral Infection of Human SCARB2-Transgenic Weaned Mice
Enterovirus A71 Induces Neurological Diseases and Dynamic Variants in Oral Infection of Human SCARB2-Transgenic Weaned Mice
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DOI：
10.1128/jvi.00897-21
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发表时间：
2021-11-01
影响因子：
5.4
通讯作者：
Shih, Shin-Ru
中科院分区：
文献类型：
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作者：
Lin, Jing-Yi;Weng, Kuo-Feng;Shih, Shin-Ru
肠道病毒A71（EV-A71）和小核糖核酸病毒科的许多成员是全球关注的嗜神经性病原体。这些病毒主要通过粪-口途径传播，因此需要合适的口腔感染动物模型来研究病毒的发病机制。使用表达人SCARB 2（hSCARB 2 Tg）、鼠适应性EV-A71/MP4病毒和具有工程化纳米荧光素酶基因的EV-A71/MP4病毒的转基因小鼠开发了口腔感染的动物模型，所述工程化纳米荧光素酶基因允许对感染小鼠中的病毒复制和传播进行成像。还对感染小鼠的组织和器官中的EV-A71基因组进行了下一代测序。口服接种EV-A71/MP4或携带纳米荧光素酶的MP4病毒可稳定诱导感染的21日龄断奶小鼠的神经系统症状和死亡。感染小鼠的体内生物发光成像和病毒抗原的组织免疫染色表明，口服接种的病毒可以传播到中枢神经系统（CNS）和其他组织。下一代测序进一步确定了病毒基因组中可能导致病毒发病机制的各种突变。本研究提出了一种EV-A71口服感染小鼠模型，该模型能够有效感染断奶小鼠，并允许跟踪病毒传播，其特征可以促进对病毒发病机制和通过自然感染途径侵袭神经的研究。重要信息肠道病毒A71（EV-A71），一种小核糖核酸病毒科的正链RNA病毒，构成了一个持续的全球公共卫生问题。EV-A71主要通过粪-口途径传播，因此需要合适的口腔感染动物模型来研究病毒的发病机制。我们提出了一种动物模型EV-A71感染，使自然途径的口腔感染断奶和非免疫功能低下的21日龄hSCARB 2转基因小鼠。我们的研究结果表明，严重的疾病和死亡可以稳定地诱导，病毒入侵的中枢神经系统可以在这个模型中复制，类似于严重的现实世界EV-A71感染。我们还开发了一种含有纳米荧光素酶的EV-A71病毒，可用于该动物模型，以真实的时间追踪口腔感染后的病毒传播。这种模型提供了优于现有动物模型的几个优点，并且可以促进未来对病毒传播、组织嗜性和病毒发病机制的研究，这些都是EV-A71感染尚未解决的紧迫问题。
Enterovirus A71 (EV-A71) and many members of the Picornaviridae family are neurotropic pathogens of global concern. These viruses are primarily transmitted through the fecal-oral route, and thus suitable animal models of oral infection are needed to investigate viral pathogenesis. An animal model of oral infection was developed using transgenic mice expressing human SCARB2 (hSCARB2 Tg), murineadapted EV-A71/MP4 virus, and EV-A71/MP4 virus with an engineered nanoluciferase gene that allows imaging of viral replication and spread in infected mice. Next-generation sequencing of EV-A71 genomes in the tissues and organs of infected mice was also performed. Oral inoculation of EV-A71/MP4 or nanoluciferase-carrying MP4 virus stably induced neurological symptoms and death in infected 21-day-old weaned mice. In vivo bioluminescence imaging of infected mice and tissue immunostaining of viral antigens indicated that orally inoculated virus can spread to the central nervous system (CNS) and other tissues. Next-generating sequencing further identified diverse mutations in viral genomes that can potentially contribute to viral pathogenesis. This study presents an EV-A71 oral infection murine model that efficiently infects weaned mice and allows tracking of viral spread, features that can facilitate research into viral pathogenesis and neuroinvasion via the natural route of infection.IMPORTANCE Enterovirus A71 (EV-A71), a positive-strand RNA virus of the Picornaviridae, poses a persistent global public health problem. EV-A71 is primarily transmitted through the fecal-oral route, and thus suitable animal models of oral infection are needed to investigate viral pathogenesis. We present an animal model of EV-A71 infection that enables the natural route of oral infection in weaned and nonimmunocompromised 21-day-old hSCARB2 transgenic mice. Our results demonstrate that severe disease and death could be stably induced, and viral invasion of the CNS could be replicated in this model, similar to severe real-world EV-A71 infections. We also developed a nanoluciferase-containing EV-A71 virus that can be used with this animal model to track viral spread after oral infection in real time. Such a model offers several advantages over existing animal models and can facilitate future research into viral spread, tissue tropism, and viral pathogenesis, all pressing issues that remain unaddressed for EV-A71 infections.