An Mx1 promoter-reporter system to study interferon pathways in rainbow trout

Dev Comp Immunol. 2004 Jun;28(7-8):793-801. doi: 10.1016/j.dci.2003.12.005.

Abstract

A rainbow trout interferon (IFN) reporter system has been established by selection of a stable cell line, RTG-P1, transfected with a plasmid expressing the firefly luciferase gene under the control of the promoter for the IFN-induced gene Mx1. After 148 passages, the luciferase expression was still highly induced by polyinosinic:polycytidylic acid (poly I:C) in RTG-P1 cells. Different IFN inducers (dsRNA, viral hemorrhagic septicaemia virus or conditioned medium containing rainbow trout antiviral activity) were able to stimulate the IFN-reporter system in RTG-P1, showing that this cell line can be used to study the activation of the IFN pathway in various contexts. Pyrrolidine dithiocarbamate (PDTC), an NF-kappaB inhibitor, significantly blocked poly I:C induced luciferase accumulation in RTG-P1 at intermediate doses (1-10 microM), suggesting that Mx1 induction through the IFN signalling pathway is NF-kappaB-dependent in fish. This inhibition was not observed for doses of 50 microM or higher. The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signalling in teleost fish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology
  • Cells, Cultured
  • Genes, Reporter / genetics
  • Interferons / genetics
  • Interferons / metabolism*
  • NF-kappaB-Inducing Kinase
  • Novirhabdovirus / immunology
  • Oncorhynchus mykiss / genetics
  • Oncorhynchus mykiss / metabolism*
  • Oncorhynchus mykiss / virology
  • Poly I-C / pharmacology
  • Promoter Regions, Genetic / genetics*
  • Protein Serine-Threonine Kinases / metabolism*
  • Pyrrolidines / pharmacology
  • Thiocarbamates / pharmacology

Substances

  • Antiviral Agents
  • Pyrrolidines
  • Thiocarbamates
  • pyrrolidine dithiocarbamic acid
  • Interferons
  • Protein Serine-Threonine Kinases
  • Poly I-C