Fu QX, Wang LC, Jia SZ, Gao B, Zhou Y, Du J, Wang YL, Wang XH, Peng JC, Zhan LS. Screening compounds against HCV based on MAVS/IFN-β pathway in a replicon model. World J Gastroenterol 2010; 16(44): 5582-5587 [PMID: 21105190 DOI: 10.3748/wjg.v16.i44.5582]
Corresponding Author of This Article
Lin-Sheng Zhan, Professor, Laboratory of Blood-Borne Virus, Beijing Institute of Transfusion Medicine, 27(9) Tai Ping Road, Beijing 100850, China. lszhan91@yahoo.com
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Original Article
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World J Gastroenterol. Nov 28, 2010; 16(44): 5582-5587 Published online Nov 28, 2010. doi: 10.3748/wjg.v16.i44.5582
Screening compounds against HCV based on MAVS/IFN-β pathway in a replicon model
Qiu-Xia Fu, Li-Cui Wang, Shuai-Zheng Jia, Bo Gao, Yong Zhou, Juan Du, Ying-Li Wang, Xiao-Hui Wang, Jian-Chun Peng, Lin-Sheng Zhan
Qiu-Xia Fu, Li-Cui Wang, Shuai-Zheng Jia, Bo Gao, Yong Zhou, Juan Du, Ying-Li Wang, Xiao-Hui Wang, Jian-Chun Peng, Lin-Sheng Zhan, Laboratory of Blood-Borne Virus, Beijing Institute of Transfusion Medicine, Beijing 100850, China
Author contributions: Zhan LS designed the research; Fu QX, Wang LC and Jia SZ performed the majority of experiments, and they contributed equally to this work; Gao B, Zhou Y, Du J, Wang YL, Wang XH and Peng JC analyzed the data and edited the manuscript.
Supported by The Natural Science Foundation of China, No. 30600330, No. 30671842, No. 30672488, No. 30700475, No. 30771919 and No. 30700757; the National High Technology Research and Development Program of China, No. 2008AA02Z132; Beijing Municipal Natural Science Foundation, No. 5082016; and Mega-projects of Science Research for the 11th Five-Year Plan, No. 2009ZX10004-4001
Correspondence to: Lin-Sheng Zhan, Professor, Laboratory of Blood-Borne Virus, Beijing Institute of Transfusion Medicine, 27(9) Tai Ping Road, Beijing 100850, China. lszhan91@yahoo.com
Telephone: +86-10-66931222 Fax: +86-10-66931292
Received: May 27, 2010 Revised: August 15, 2010 Accepted: August 23, 2010 Published online: November 28, 2010
Abstract
AIM: To develop a sensitive assay for screening compounds against hepatitis C virus (HCV).
METHODS: The proteolytic cleavage of NS3/4A on enhanced yellow fluorescent protein (eYFP)-mitochondrial antiviral signaling protein (MAVS) was examined by reporter enzyme secreted placental alkaline phosphatase (SEAP), which enabled us to perform ongoing monitoring of anti-HCV drugs through repeated chemiluminescence. Subcellular localization of eYFP-MAVS was assessed by fluorescence microscopy. Cellular localization and protein levels were examined by Western blotting.
RESULTS: HCV NS3/4A protease cleaved eYFP-MAVS from mitochondria to block the activation of interferon (IFN)-β promoter, thus resulting in downregulation of SEAP activity. The decrease in SEAP activity was proportional to the dose of active NS3/4A protease. Also this reporter assay was used to detect anti-HCV activity of IFN-α and cyclosporine A.
CONCLUSION: Our data show that this reporter system is a sensitive and quantitative reporter of anti-HCV inhibitors. This system will constitute a new tool to allow the efficient screening of HCV inhibitors.
