title: An in vitro model of latency and reactivation of varicella zoster virus in human stem cell-derived neurons.
abstract: Varicella zoster virus (VZV) latency in sensory and autonomic neurons has remained enigmatic and difficult to study, and experimental reactivation has not yet been achieved. We have previously shown that human embryonic stem cell (hESC)-derived neurons are permissive to a productive and spreading VZV infection. We now demonstrate that hESC-derived neurons can also host a persistent non-productive infection lasting for weeks which can subsequently be reactivated by multiple experimental stimuli. Quiescent infections were established by exposing neurons to low titer cell-free VZV either by using acyclovir or by infection of axons in compartmented microfluidic chambers without acyclovir. VZV DNA and low levels of viral transcription were detectable by qPCR for up to seven weeks. Quiescently-infected human neuronal cultures were induced to undergo renewed viral gene and protein expression by growth factor removal or by inhibition of PI3-Kinase activity. Strikingly, incubation of cultures induced to reactivate at a lower temperature (34°C) resulted in enhanced VZV reactivation, resulting in spreading, productive infections. Comparison of VZV genome transcription in quiescently-infected to productively-infected neurons using RNASeq revealed preferential transcription from specific genome regions, especially the duplicated regions. These experiments establish a powerful new system for modeling the VZV latent state, and reveal a potential role for temperature in VZV reactivation and disease.
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
authors.forename: Amos
authors.initials: A
authors.lastname: Markus
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
authors.forename: Ilana
authors.initials: I
authors.lastname: Lebenthal-Loinger
authors.affiliation: Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America; Singapore Institute for Neurotechnology (SINAPSE), National University of Singapore, Singapore.
authors.forename: In Hong
authors.initials: IH
authors.lastname: Yang
authors.affiliation: Department of Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America; Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Kinchington
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
authors.forename: Ronald S
authors.initials: RS
authors.lastname: Goldstein
mesh_terms: D053595:Embryonic Stem Cells / Q000821:virology; D006562:Herpes Zoster / Q000821:virology; D014645:Herpesvirus 3, Human / Q000502:physiology*; D006801:Humans; D017403:In Situ Hybridization; D066298:In Vitro Techniques; D058953:Neural Stem Cells / Q000821:virology*; D009474:Neurons / Q000821:virology*; D016133:Polymerase Chain Reaction; D014775:Virus Activation / Q000502:physiology*; D017735:Virus Latency / Q000502:physiology*