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Effect of common carp (Cyprinus carpio) TLR9 overexpression on the expression of downstream interferon-associated immune factor mRNAs in epithelioma papulosum cyprini cells.

Authors:
Wei Xinxian Jia Peng Tong Guixiang Wang Jinjin Zheng Xiaocong He Junqiang Yang Xianle Liu Hong

Vet Immunol Immunopathol 2016 Feb 23;170:47-53. Epub 2015 Oct 23.

Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518045, China. Electronic address:

Toll-like receptors (TLRs) are a class of pattern recognition receptors (PRRs) that recognize pathogen associated molecular patterns (PAMPs) and play an important role in the antiviral response. To determine the effect of common carp TLR9 (CcTLR9) overexpression on the expression of down-stream interferon associated immune factors in epithelioma papulosum cyprini (EPC) cells, may provide useful information for the further investigation on the anti-virus effect mediated by TLR9 in fish. In this study, we constructed an overexpression vector, pEGFP-N1-CcTLR9, by cloning the CcTLR9 gene and inserting it into an expression vector pEGFP-N1. Both plasmids DNA of pEGFP-N1 and pEGFP-N1-CcTLR9 were transfected into EPC cells, and the expression of IRF3, IRF7, ISG15, Mx1, PKR and Viperin mRNA at 0, 6, 12, 24, 48 and 72h post-transfection were determined by real-time quantitative PCR (Q-PCR). Overexpression of the CcTLR9 gene in EPC cells upregulates the expression of IRF3, IRF7, ISG15, Mx1, PKR, and Viperin mRNA, and this was more significant for Viperin, ISG15, and IRF7, and least significant for PKR. Thus, fish TLR9 activates IRF7 signaling to induce I-IFN secretion and the subsequent upregulation of IFN-stimulated genes.

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http://dx.doi.org/10.1016/j.vetimm.2015.10.006DOI Listing
February 2016

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