Varicella-Zoster Virus (VZV) Infection of Neurons Derived from Human Embryonic Stem Cells: Direct Demonstration of Axonal Infection, Transport of VZV, and Productive Neuronal Infection
Varicella-Zoster Virus (VZV) Infection of Neurons Derived from Human Embryonic Stem Cells: Direct Demonstration of Axonal Infection, Transport of VZV, and Productive Neuronal Infection
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DOI：
10.1128/jvi.02396-10
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发表时间：
2011-07-01
影响因子：
5.4
通讯作者：
Goldstein, Ronald S.
中科院分区：
文献类型：
--
作者：
Markus, Amos;Grigoryan, Sergei;Goldstein, Ronald S.
人类神经营养性疱疹病毒水痘带状疱疹病毒（VZV）及其感染神经元能力的研究受到严格的病毒向人类趋向性和用于实验的人类神经元有限的限制。人类胚胎干细胞（hESC）可以分化为包括神经元在内的身体所有细胞类型，因此是研究人类神经元与人类嗜神经病毒相互作用的潜在无限来源。我们在这里报告了表达细胞相关绿色荧光蛋白（GFP）的 VZV 对 hESC 衍生神经元的可重复感染。 hESC衍生的神经元在与感染重组VZV的有丝分裂抑制的MeWo细胞孵育后2天内表达GFP，所述重组VZV表达GFP作为与VZV蛋白的GFP融合物或在独立启动子下。通过立即早期和病毒衣壳蛋白的免疫染色证实了水痘带状疱疹病毒感染。 hESC 衍生的神经元的感染是富有成效的，导致释放到感染性病毒颗粒的介质中，在电子显微镜下观察到这些病毒颗粒似乎已完全组装。我们还首次证明了 VZV 对轴突的感染以及使用隔室微流体室从轴突到神经元细胞体的逆行运输。将 hESC 衍生的人类神经元与荧光标记的 VZV 结合使用，为研究 VZV 神经元感染和轴突运输显示出巨大的前景，并有可能建立人类神经元中 VZV 潜伏期的模型。
Study of the human neurotrophic herpesvirus varicella-zoster virus (VZV) and of its ability to infect neurons has been severely limited by strict viral human tropism and limited availability of human neurons for experimentation. Human embryonic stem cells (hESC) can be differentiated to all the cell types of the body including neurons and are therefore a potentially unlimited source of human neurons to study their interactions with human neurotropic viruses. We report here reproducible infection of hESC-derived neurons by cell-associated green fluorescent protein (GFP)-expressing VZV. hESC-derived neurons expressed GFP within 2 days after incubation with mitotically inhibited MeWo cells infected with recombinant VZV expressing GFP as GFP fusions to VZV proteins or under an independent promoter. VZV infection was confirmed by immunostaining for immediate-early and viral capsid proteins. Infection of hESC-derived neurons was productive, resulting in release into the medium of infectious virions that appeared fully assembled when observed by electron microscopy. We also demonstrated, for the first time, VZV infection of axons and retrograde transport from axons to neuronal cell bodies using compartmented microfluidic chambers. The use of hESC-derived human neurons in conjunction with fluorescently tagged VZV shows great promise for the study of VZV neuronal infection and axonal transport and has potential for the establishment of a model for VZV latency in human neurons.