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Completion of the entire hepatitis C virus life cycle in genetically humanized mice


More than 130 million people worldwide chronically infected with hepatitis C virus (HCV) are at risk of developing severe liver disease. Antiviral treatments are only partially effective against HCV infection, and a vaccine is not available. Development of more efficient therapies has been hampered by the lack of a small animal model. Building on the observation that CD81 and occludin (OCLN) comprise the minimal set of human factors required to render mouse cells permissive to HCV entry1, we previously showed that transient expression of these two human genes is sufficient to allow viral uptake into fully immunocompetent inbred mice2. Here we demonstrate that transgenic mice stably expressing human CD81 and OCLN also support HCV entry, but innate and adaptive immune responses restrict HCV infection in vivo. Blunting antiviral immunity in genetically humanized mice infected with HCV results in measurable viraemia over several weeks. In mice lacking the essential cellular co-factor cyclophilin A (CypA), HCV RNA replication is markedly diminished, providing genetic evidence that this process is faithfully recapitulated. Using a cell-based fluorescent reporter activated by the NS3-4A protease we visualize HCV infection in single hepatocytes in vivo. Persistently infected mice produce de novo infectious particles, which can be inhibited with directly acting antiviral drug treatment, thereby providing evidence for the completion of the entire HCV life cycle in inbred mice. This genetically humanized mouse model opens new opportunities to dissect genetically HCV infection in vivo and provides an important preclinical platform for testing and prioritizing drug candidates and may also have utility for evaluating vaccine efficacy.

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Figure 1: Transgenic expression of human CD81 and OCLN renders mice permissive to HCV entry.
Figure 2: Blunting of antiviral immune responses in mice expressing HCV entry factors augments HCV RNA replication.
Figure 3: Visualization and genetic and pharmacological interference with HCV infection.
Figure 4: HCV infection in 4hEF Stat1−/− mice leads to immune activation.
Figure 5: Evidence for production of infectious particles.


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We thank J. Sable, E. Castillo, B. Flatley, S. Shirley, A. Webson and E. Giang for laboratory support. A. North and the Rockefeller University Bioimaging Core Facility, S. Mazel and the Rockefeller University Flowcytometry Core Facility, C. Yang and the Gene Targeting Center and R. Tolwani and the staff of the Comparative Bioscience Center provided technical support. This study was supported in part by award number RC1DK087193 (to C.M.R. and A.P.) from the National Institute of Diabetes and Digestive and Kidney Diseases, R01AI072613, R01AI099284 (to C.M.R.), R01AI079031 (to M.L.) from the National Institute for Allergy and Infectious Disease, R01CA057973 (to C.M.R.) from the National Cancer Institute, The Starr Foundation, the Greenberg Medical Research Institute, the Richard Salomon Family Foundation, the Ronald A. Shellow, M.D. Memorial Fund, the MGM Mirage Voice Foundation, Gregory F. Lloyd Memorial contributions, and anonymous donors. M.D. was supported by a postdoctoral fellowship from the German Research Foundation (Deutsche Forschungsgesellschaft). M.T.C. is a recipient of The Rockefeller University Women & Science Fellowship. A.P. is a recipient of the Astella Young Investigator Award from the Infectious Disease Society of America and a Liver Scholar Award from the American Liver Foundation. The funding sources were not involved in the study design, collection, analysis and interpretation of data or in the writing of the report.

Author information




M.D. and J.A.H. planned and performed experiments and contributed to writing the manuscript. B.M.D., R.N.L., W.C.B., T.F., A.V. and M.T.C. performed the experimental work; T.K., T.S., S.A. and M.L. provided reagents. C.M.R. provided laboratory infrastructure, space, reagents, advice and edited the manuscript. A.P. planned and performed experiments and wrote the manuscript.

Corresponding authors

Correspondence to Charles M. Rice or Alexander Ploss.

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Competing interests

The following conflicts of interest are managed under University policy: C.M.R. has equity in Apath, LLC, which holds commercial licenses for the Huh-7.5 cell line, HCV cell culture system, the use of OCLN to construct HCV animal models and the fluorescent cell-based reporter system to detect HCV infection.

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Dorner, M., Horwitz, J., Donovan, B. et al. Completion of the entire hepatitis C virus life cycle in genetically humanized mice. Nature 501, 237–241 (2013).

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