Infection of primary human hepatocytes with recombinant hepatitis C virus

Attachment of the virus to the cell surface followed by viral entry is the first step in a cascade of events required for initiation of infection. Results obtained with surrogate model systems for HCV infection using human hepatoma cell lines suggest that HCV entry is most likely a multi-step process which requires a set of initial binding molecules such as glycosaminoglycans, as well as (co)-receptors for viral entry such as CD81, SR-BI and claudin–1. However, it still remains elusive which host cell factors are required for HCV entry into its natural target cell, the human hepatocyte. In this study, we used the recently established HCV-JFH–1 based-infectious tissue culture model system to generate high titer virion preparations (HCVcc) for infection of primary human hepatocytes (PHH). Cells were incubated with cell culture adapted JFH–1 with proven in vivo viability in the presence or absence of an NS3 protease inhibitor. Infection was assessed by measuring HCV RNA by quantitative RT-PCR at different time-points post infection. PHH could be reproducibly infected with HCVcc as shown by a time-dependent increase of HCV RNA in infected hepatocytes which was not observed when cells were treated with the NS3 protease inhibitor. Studies to assess the impact of CD81, SR-BI and claudin–1 as host entry factor for HCV infection of primary human hepatocytes are ongoing. In conclusion, our results suggest that PHH can be infected with cell culture grown HCV. This model system will allow further dissecting molecular mechanisms of HCV infection of its natural target cell – the human hepatocyte. cell culture-derived HCV - hepatitis c virus - primary human hepatocytes