title: Effect of common carp (Cyprinus carpio) TLR9 overexpression on the expression of downstream interferon-associated immune factor mRNAs in epithelioma papulosum cyprini cells.
abstract: 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.
authors: Shanghai Ocean University, College of Fisheries and Life Science, Shanghai 201306, China; Guangxi Academy of Fishery Science, Nanning 530021, China.
authors: Wei
authors: W
authors: Xinxian
authors: Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518045, China.
authors: Jia
authors: J
authors: Peng
authors: Guangxi Academy of Fishery Science, Nanning 530021, China.
authors: Tong
authors: T
authors: Guixiang
authors: Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518045, China.
authors: Wang
authors: W
authors: Jinjin
authors: Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518045, China.
authors: Zheng
authors: Z
authors: Xiaocong
authors: Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518045, China.
authors: He
authors: H
authors: Junqiang
authors: Shanghai Ocean University, College of Fisheries and Life Science, Shanghai 201306, China. Electronic address: xlyang@shou.edu.cn.
authors: Yang
authors: Y
authors: Xianle
authors: Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518045, China. Electronic address: liuhong@szciq.gov.cn.
authors: Liu
authors: L
authors: Hong
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002347:Carps / Q000276:immunology*; D002460:Cell Line; D005393:Fish Diseases / Q000276:immunology* / Q000821:virology; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D007372:Interferons / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis; D051217:Toll-Like Receptor 9 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D014162:Transfection
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger; D051217:Toll-Like Receptor 9; D007372:Interferons
keywords: IFN-stimulated genes; Interferon regulatory factor; Overexpression; TLR9; mRNA expression