Species-independent bioassay for sensitive quantification of antiviral type I interferons

Virol J. 2010 Feb 26:7:50. doi: 10.1186/1743-422X-7-50.

Abstract

Background: Studies of the host response to infection often require quantitative measurement of the antiviral type I interferons (IFN-alpha/beta) in biological samples. The amount of IFN is either determined via its ability to suppress a sensitive indicator virus, by an IFN-responding reporter cell line, or by ELISA. These assays however are either time-consuming and lack convenient readouts, or they are rather insensitive and restricted to IFN from a particular host species.

Results: An IFN-sensitive, Renilla luciferase-expressing Rift Valley fever virus (RVFV-Ren) was generated using reverse genetics. Human, murine and avian cells were tested for their susceptibility to RVFV-Ren after treatment with species-specific IFNs. RVFV-Ren was able to infect cells of all three species, and IFN-mediated inhibition of viral reporter activity occurred in a dose-dependent manner. The sensitivity limit was found to be 1 U/ml IFN, and comparison with a standard curve allowed to determine the activity of an unknown sample.

Conclusions: RVFV-Ren replicates in cells of several species and is highly sensitive to pre-treatment with IFN. These properties allowed the development of a rapid, sensitive, and species-independent antiviral assay with a convenient luciferase-based readout.

Publication types

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

MeSH terms

  • Animals
  • Biological Assay / methods*
  • Birds
  • Cell Line
  • Genes, Reporter
  • Humans
  • Interferon Type I / immunology*
  • Luciferases, Renilla / genetics
  • Luciferases, Renilla / metabolism
  • Mice
  • Sensitivity and Specificity
  • Virus Diseases / immunology*
  • Viruses / immunology*

Substances

  • Interferon Type I
  • Luciferases, Renilla