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

B Collet, P Boudinot, A Benmansour, C J Secombes

Research output: Contribution to journalArticle

70 Citations (Scopus)

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 muM), suggesting that Mx1 induction through the IFN signalling pathway is NF-kappaB-dependent in fish. This inhibition was not observed for doses of 50 muM or higher. The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signalling in teleost fish. (C) 2004 Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)793-801
Number of pages9
JournalDevelopmental and Comparative Immunology
Volume28
DOIs
Publication statusPublished - 2004

Keywords

  • NF-KAPPA-B
  • DOUBLE-STRANDED-RNA
  • HEMORRHAGIC SEPTICEMIA VIRUS
  • VESICULAR STOMATITIS-VIRUS
  • MOLECULAR-CLONING
  • ATLANTIC SALMON
  • INFLUENZA-VIRUS
  • EXPRESSION
  • GENE
  • CELLS

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