Titration of cell-associated varicella-zoster virus with the MV9G reporter cell line for antiviral studies

J Virol Methods. 2018 Oct:260:14-20. doi: 10.1016/j.jviromet.2018.06.019. Epub 2018 Jun 30.

Abstract

Titration of the cell-associated virus (CAV) of varicella-zoster virus (VZV) is essential for antiviral studies. A VZV reporter cell line, MV9G, generated in our previous study expresses firefly luciferase upon CAV infection in a dose-dependent manner, suggesting that use of the cell line for titration is feasible. In this study, MeWo cells infected with VZV vaccine Oka (vOka) strain or with clinical isolates obtained from patients with varicella or zoster were used as CAV. A co-culture of MV9G cells with the virus-infected MeWo cells were set up and optimized for titration of CAV. Luciferase activities of MV9G cells measured as relative light units (RLUs) of chemiluminescence correlated well (r > 0.9, p < 0.05) both with quantities of viral DNAs measured by TaqMan PCR and with numbers of viral foci detected by immunostaining with a monoclonal antibody against VZV IE62. In addition, the usefulness of MV9G for antiviral studies was exemplified by treatment of the VZV-infected cells with various concentrations of acyclovir. Thus, the reporter cell-based titration of CAV by measuring the induced RLUs may be a reliable way to estimate viral foci and viral DNAs.

Keywords: Cell-associated virus; Reporter cell line; Titration; Varicella-zoster virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyclovir / pharmacology
  • Antiviral Agents / pharmacology
  • Cell Line
  • Cell Line, Tumor
  • Coculture Techniques
  • DNA, Viral / drug effects
  • DNA, Viral / isolation & purification*
  • Genes, Reporter / genetics*
  • Herpesvirus 3, Human / drug effects
  • Herpesvirus 3, Human / enzymology*
  • Humans
  • Luciferases, Firefly / genetics
  • Luminescence
  • Varicella Zoster Virus Infection / virology*
  • Viral Load / genetics*

Substances

  • Antiviral Agents
  • DNA, Viral
  • Luciferases, Firefly
  • Acyclovir