Modeling HSV-1 Latency in Human Embryonic Stem Cell-Derived Neurons.
Modeling HSV-1 Latency in Human Embryonic Stem Cell-Derived Neurons.
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DOI：
10.3390/pathogens6020024
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发表时间：
2017-06-08
期刊：
影响因子：
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通讯作者：
Wilson AC
中科院分区：
文献类型：
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作者：
Pourchet A;Modrek AS;Placantonakis DG;Mohr I;Wilson AC
单纯疱疹病毒1型(HSV-1)利用外周神经节的潜伏期在其宿主体内存活，然而，从这个储存库反复重新激活可能会导致虚弱和潜在威胁生命的疾病。大多数关于潜伏期的研究使用活体动物感染模型，但这些模型是复杂的、多层次的系统，可能很难操作。感染培养的原代神经元提供了一个强大的替代方案，为控制潜伏期的宿主信号通路提供了重要的见解。然而，小动物模型并不能概括人类HSV-1感染的所有方面，而且在可用的分子工具方面也是有限的。为了解决这个问题，我们开发了一个潜伏期模型，该模型基于培养中从NIH批准的胚胎干细胞系分化出的人类神经元。由此产生的神经元高度允许复制野生型单纯疱疹病毒1型，但建立了一种非生产性感染状态，类似于在抗病毒药物阿昔洛韦和干扰素-α存在的情况下以低病毒剂量感染时的潜伏期。在这种状态下，没有检测到病毒复制和晚期病毒基因标记的表达，但有病毒潜伏期相关转录本(LAT)RNA的积累。在六天的建制期后，可以移除抗病毒药物，并将受感染的培养保持几周。随后用丁酸钠治疗会导致新的传染性病毒的重新激活和产生。来自干细胞的人类神经元为研究这种专门针对人类的病毒提供了适当的物种背景，有可能对祖细胞进行更广泛的操纵，并获得广泛的先前存在的分子工具。
Herpes simplex virus 1 (HSV-1) uses latency in peripheral ganglia to persist in its human host, however, recurrent reactivation from this reservoir can cause debilitating and potentially life-threatening disease. Most studies of latency use live-animal infection models, but these are complex, multilayered systems and can be difficult to manipulate. Infection of cultured primary neurons provides a powerful alternative, yielding important insights into host signaling pathways controlling latency. However, small animal models do not recapitulate all aspects of HSV-1 infection in humans and are limited in terms of the available molecular tools. To address this, we have developed a latency model based on human neurons differentiated in culture from an NIH-approved embryonic stem cell line. The resulting neurons are highly permissive for replication of wild-type HSV-1, but establish a non-productive infection state resembling latency when infected at low viral doses in the presence of the antivirals acyclovir and interferon-α. In this state, viral replication and expression of a late viral gene marker are not detected but there is an accumulation of the viral latency-associated transcript (LAT) RNA. After a six-day establishment period, antivirals can be removed and the infected cultures maintained for several weeks. Subsequent treatment with sodium butyrate induces reactivation and production of new infectious virus. Human neurons derived from stem cells provide the appropriate species context to study this exclusively human virus with the potential for more extensive manipulation of the progenitors and access to a wide range of preexisting molecular tools.