title: A quantitative infection assay for human type I, II, and III interferon antiviral activities.
abstract: BACKGROUND
  Upon virus infection, cells secrete a diverse group of antiviral molecules that signal proximal cells to enter into an antiviral state, slowing or preventing viral spread. These paracrine signaling molecules can work synergistically, so measurement of any one antiviral molecule does not reflect the total antiviral activity of the system.
  
  
  RESULTS
  We have developed an antiviral assay based on replication inhibition of an engineered fluorescent vesicular stomatitis virus reporter strain on A549 human lung epithelial cells. Our assay provides a quantitative functional readout of human type I, II, and III interferon activities, and it provides better sensitivity, intra-, and inter-assay reproducibility than the traditional crystal violet based assay. Further, it eliminates cell fixation, rinsing, and staining steps, and is inexpensive to implement.
  
  
  CONCLUSIONS
  A dsRed2-strain of vesicular stomatitis virus that is sensitive to type I, II, and III interferons was used to develop a convenient and sensitive assay for interferon antiviral activity. We demonstrate use of the assay to quantify the kinetics of paracrine antiviral signaling from human prostate cancer (PC3) cells in response to viral infection. The assay is applicable to high-throughput screening for anti-viral compounds as well as basic studies of cellular antiviral signaling.
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin, Madison, USA.
authors.forename: Emily
authors.initials: E
authors.lastname: Voigt
authors.forename: Bahar
authors.initials: B
authors.lastname: Inankur
authors.forename: Ashley
authors.initials: A
authors.lastname: Baltes
authors.forename: John
authors.initials: J
authors.lastname: Yin
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D004847:Epithelial Cells / Q000821:virology; D005453:Fluorescence; D017930:Genes, Reporter; D006801:Humans; D007372:Interferons / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests / Q000379:methods; D018116:Vesiculovirus / Q000187:drug effects* / Q000235:genetics
chemical_list: D000998:Antiviral Agents; D007372:Interferons