title: In vitro system using human neurons demonstrates that varicella-zoster vaccine virus is impaired for reactivation, but not latency.
abstract: Varicella-zoster virus (VZV) establishes latency in human sensory and cranial nerve ganglia during primary infection (varicella), and the virus can reactivate and cause zoster after primary infection. The mechanism of how the virus establishes and maintains latency and how it reactivates is poorly understood, largely due to the lack of robust models. We found that axonal infection of neurons derived from hESCs in a microfluidic device with cell-free parental Oka (POka) VZV resulted in latent infection with inability to detect several viral mRNAs by reverse transcriptase-quantitative PCR, no production of infectious virus, and maintenance of the viral DNA genome in endless configuration, consistent with an episome configuration. With deep sequencing, however, multiple viral mRNAs were detected. Treatment of the latently infected neurons with Ab to NGF resulted in production of infectious virus in about 25% of the latently infected cultures. Axonal infection of neurons with vaccine Oka (VOka) VZV resulted in a latent infection similar to infection with POka; however, in contrast to POka, VOka-infected neurons were markedly impaired for reactivation after treatment with Ab to NGF. In addition, viral transcription was markedly reduced in neurons latently infected with VOka compared with POka. Our in vitro system recapitulates both VZV latency and reactivation in vivo and may be used to study viral vaccines for their ability to establish latency and reactivate.
authors.affiliation: Medical Virology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892;
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Sadaoka
authors.affiliation: Division of Infection and Immunity, MRC Centre for Medical Molecular Virology, University College London, London WC1E 6BT, United Kingdom;
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Depledge
authors.affiliation: Division of Neuroimmunology and Neuroinfectious Diseases, Department of Neurology, Johns Hopkins University School of Medicine, Johns Hopkins Hospital, Baltimore, MD 21287.
authors.forename: Labchan
authors.initials: L
authors.lastname: Rajbhandari
authors.affiliation: Division of Neuroimmunology and Neuroinfectious Diseases, Department of Neurology, Johns Hopkins University School of Medicine, Johns Hopkins Hospital, Baltimore, MD 21287.
authors.forename: Arun
authors.initials: A
authors.lastname: Venkatesan
authors.affiliation: Division of Infection and Immunity, MRC Centre for Medical Molecular Virology, University College London, London WC1E 6BT, United Kingdom;
authors.forename: Judith
authors.initials: J
authors.lastname: Breuer
authors.affiliation: Medical Virology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; jcohen@niaid.nih.gov.
authors.forename: Jeffrey I
authors.initials: JI
authors.lastname: Cohen
mesh_terms: D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006562:Herpes Zoster / Q000517:prevention & control / Q000821:virology; D053061:Herpes Zoster Vaccine / Q000494:pharmacology*; D014645:Herpesvirus 3, Human / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D044085:Microfluidics; D058953:Neural Stem Cells / Q000821:virology*; D014775:Virus Activation / Q000502:physiology; D017735:Virus Latency / Q000502:physiology
chemical_list: D053061:Herpes Zoster Vaccine
keywords: herpesvirus; latency; reactivation; varicella vaccine; varicella-zoster virus