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Updated: Jun 4, 2026
A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Studies on virus kinetics using infectious fluorescence-tagged hepatitis C virus cell culture
Machi Yamamoto1, Naoya Sakamoto, Tetsuya Nakamura
1Department of Gastroenterology and Hepatology Department for Hepatitis Control Department of Advanced Therapeutics in Gastrointestinal Diseases, Tokyo Medical and Dental University Department of Virology II, National Institute of Infectious Disease, Tokyo Department of Internal Medicine, Soka Municipal Hospital, Saitama, Japan.
Aim:
Studies of the complete hepatitis C virus (HCV) life cycle have become possible with the development of a HCV-JFH1 cell culture system.
Methods:
In this study, we constructed two fluorescence protein-tagged recombinant JFH1 virus clones, JFH1-EYFP and JFH1-AsRed, as well as two corresponding clones with adaptive mutations, JFH1-EYFP mutant and JFH1-AsRed mutant, that and were as effective as JFH1 in producing infectious virus particles, and investigated their viral infection life cycles.
Results:
After infection of the fluorescence-tagged mutant viruses, infected cells increased exponentially. In cells, EYFP or AsRed and NS5A were expressed as a fusion protein and co-localized in core proteins. The rate of the cell-cell spread was dependent on the cell densities with a maximum of 10(2.5) /day. Treatment of cells with interferon or a protease inhibitor suppressed expansion of virus-positive cells.
Conclusion:
Taken together, these results indicate that fluorescence-tagged HCV is a useful tool to study virus infection life cycles and to assist in the search for novel antiviral compounds.
