Herpes simplex virus (HSV) immediate-early (IE) promoter-directed reporter system for the screening of antiherpetics targeting the early stage of HSV infection

J Biomol Screen. 2010 Sep;15(8):1016-20. doi: 10.1177/1087057110372804. Epub 2010 Jun 14.

Abstract

Most of the current antiherpetics target viral DNA polymerase, but with the emergence of drug-resistant viruses, antiherpetics with different targets have become necessary. Inhibition of herpes simplex virus (HSV) replication at the early stages of infection minimizes cytotoxicity and immune suppression induced by HSV infection. In this report, quantitative reporter systems that use recombinant HSV and a stably transfected cell line were developed for the screening of agents targeting the early stages of HSV infection. The reporter genes in both systems were directed by HSV immediate-early (IE) promoters, so considerably less time was required for the quantification of HSV infection than the traditional plaque reduction assay. The results show that both reporter assays were sensitive to antiherpetic screening. Both assays were quantitative, rapid, easy to perform, and highly adaptable for automatic high-throughput screening. Exploiting the flexibility of these 2 assays, modified assays were also proposed for the detailed analysis of antiherpetic mechanisms.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Antiviral Agents / analysis
  • Antiviral Agents / isolation & purification*
  • Antiviral Agents / metabolism
  • Antiviral Agents / pharmacology
  • Cells, Cultured
  • Chlorocebus aethiops
  • Disease Progression
  • Gene Expression Regulation, Viral
  • Genes, Reporter
  • Herpes Simplex / genetics*
  • Herpes Simplex / metabolism
  • Herpes Simplex / pathology
  • Immediate-Early Proteins / genetics*
  • Immediate-Early Proteins / metabolism*
  • Molecular Targeted Therapy / methods
  • Promoter Regions, Genetic* / genetics
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Simplexvirus / genetics*
  • Transfection
  • Vero Cells

Substances

  • Antiviral Agents
  • Immediate-Early Proteins
  • Recombinant Fusion Proteins