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

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-kB inhibitor, significantly blocked poly I:C induced luciferase accumulation in RTG-P1 at intermediate doses (1–10 mM), suggesting that Mx1 induction through the IFN signalling pathway is NF-kB-dependent in fish. This inhibition was not observed for doses of 50 mMor higher. The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signalling in teleost fish.
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Submitted on : Monday, June 1, 2020 - 2:27:41 AM
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Bertrand Collet, Pierre Boudinot, Abdenour Benmansour, Christopher J. Secombes. An Mx1 promoter-reporter system to study interferon pathways in rainbow trout. Developmental and Comparative Immunology, Elsevier, 2004, 28 (7-8), pp.793-801. ⟨10.1016/j.dci.2003.12.005⟩. ⟨hal-02683144⟩



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