A Luciferase Reporter Gene Assay to Measure Ebola Virus Viral Protein 35-Associated Inhibition of Double-Stranded RNA-Stimulated, Retinoic Acid-Inducible Gene 1-Mediated Induction of Interferon β

J Infect Dis. 2015 Oct 1:212 Suppl 2:S277-81. doi: 10.1093/infdis/jiv214. Epub 2015 Apr 29.

Abstract

During Ebola virus (EBOV) infection, the type I interferon α/β (IFN-α/β) innate immune response is suppressed by EBOV viral protein 35 (VP35), a validated drug target. Identification of EBOV VP35 inhibitors requires a cellular system able to assess the VP35-based inhibitory functions of viral double-stranded RNA (dsRNA) IFN-β induction. We established a miniaturized luciferase gene reporter assay in A549 cells that measures IFN-β induction by viral dsRNA and is dose-dependently inhibited by VP35 expression. When compared to influenza A virus NS1 protein, EBOV VP35 showed improved inhibition of viral dsRNA-based IFN-β induction. This assay can be used to screen for EBOV VP35 inhibitors.

Keywords: Ebola virus; IFN-β induction; IFN-β inhibition; NS1; RIG-I activation; VP35; drug development; dsRNA recognition; innate immunity; luciferase reporter gene assay.

MeSH terms

  • Cell Line
  • Ebolavirus / genetics*
  • Genes, Reporter / genetics*
  • Hemorrhagic Fever, Ebola / genetics*
  • Hemorrhagic Fever, Ebola / virology
  • Humans
  • Interferon-alpha / genetics
  • Interferon-beta / genetics*
  • Luciferases / metabolism*
  • RNA, Double-Stranded / genetics*
  • Transcriptional Activation / genetics
  • Tretinoin / metabolism
  • Viral Nonstructural Proteins / genetics
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • INS1 protein, influenza virus
  • Interferon-alpha
  • RNA, Double-Stranded
  • Viral Nonstructural Proteins
  • Viral Proteins
  • Tretinoin
  • Interferon-beta
  • Luciferases