title: 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.
abstract: 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.
authors.affiliation: Mina and Everard Goodman Faculty of Life Sciences, Gonda Building, Old Campus, 52900 Ramat-Gan, Israel.
authors.forename: Amos
authors.initials: A
authors.lastname: Markus
authors.forename: Sergei
authors.initials: S
authors.lastname: Grigoryan
authors.forename: Anna
authors.initials: A
authors.lastname: Sloutskin
authors.forename: Michael B
authors.initials: MB
authors.lastname: Yee
authors.forename: Hua
authors.initials: H
authors.lastname: Zhu
authors.forename: In Hong
authors.initials: IH
authors.lastname: Yang
authors.forename: Nitish V
authors.initials: NV
authors.lastname: Thakor
authors.forename: Ronit
authors.initials: R
authors.lastname: Sarid
authors.forename: Paul R
authors.initials: PR
authors.lastname: Kinchington
authors.forename: Ronald S
authors.initials: RS
authors.lastname: Goldstein
mesh_terms: D001370:Axonal Transport; D001369:Axons / Q000648:ultrastructure / Q000821:virology*; D002213:Capsid / Q000378:metabolism / Q000648:ultrastructure; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D006801:Humans; D046529:Microscopy, Electron, Transmission; D009474:Neurons / Q000648:ultrastructure / Q000821:virology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure; D057074:Virus Release
chemical_list: D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins