An Mx1 promoter-reporter system to study interferon pathways in rainbow trout.
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.
Collet
Bertrand
B
Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Zoology Building, Tillydrone Avenue, Aberdeen AB24 2TZ, Scotland, UK.
Boudinot
Pierre
P
Benmansour
Abdenour
A
Secombes
Christopher J
CJ
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D017930:Genes, Reporter / Q000235:genetics; D007372:Interferons / Q000235:genetics / Q000378:metabolism*; D029141:Novirhabdovirus / Q000276:immunology; D017686:Oncorhynchus mykiss / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D011070:Poly I-C / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011759:Pyrrolidines / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D000097782:NF-kappaB-Inducing Kinase
D000998:Antiviral Agents; D007372:Interferons; D011070:Poly I-C; D017346:Protein Serine-Threonine Kinases; D011759:Pyrrolidines; D013859:Thiocarbamates; D000097782:NF-kappaB-Inducing Kinase; C020972:pyrrolidine dithiocarbamic acid