Creation and characterization of a cell-death reporter cell line for hepatitis C virus infection.
The present study describes the creation and characterization of a hepatoma cell line, n4mBid, that supports all stages of the hepatitis C virus (HCV) life cycle and strongly reports HCV infection by a cell-death phenotype. The n4mBid cell line is derived from the highly HCV-permissive Huh-7.5 hepatoma cell line and contains a modified Bid protein (mBid) that is cleaved and activated by the HCV serine protease NS3-4A. N4mBid exhibited a 10-20-fold difference in cell viability between the HCV-infected and mock-infected states, while the parental Huh-7.5 cells showed <2-fold difference under the same conditions. The pronounced difference in n4mBid cell viability between the HCV- and mock-infected states in a 96-well plate format points to its usefulness in cell survival-based high-throughput screens for anti-HCV molecules. The degree of cell death was found to be proportional to the intracellular load of HCV. HCV-low n4mBid cells, expressing an anti-HCV short hairpin RNA, showed a significant growth advantage over naïve cells and could be rapidly enriched after HCV infection, suggesting the possibility of using n4mBid cells for the cell survival-based selection of genetic anti-HCV factors.
Chen
Zhilei
Z
Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065, USA. zhilei.chen@chemail.tamu.edu
Simeon
Rudo L
RL
Chockalingam
Karuppiah
K
Rice
Charles M
CM
D018929:Cell Culture Techniques / Q000379:methods; D016923:Cell Death; D045744:Cell Line, Tumor; D002470:Cell Survival; D016174:Hepacivirus / Q000254:growth & development / Q000472:pathogenicity*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D014773:Virology / Q000379:methods*