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Entry of hepatitis C virus pseudotypes into primary human hepatocytes by clathrin-dependent endocytosis

Audrey Codran^1,

¹ INSERM U544, EA 3770, Institut de Virologie, Université Louis Pasteur, 3 rue Koeberlé, F-67000 Strasbourg, France
² Centre de Chirurgie Viscérale et de Transplantation, Centre Hospitalier de Hautepierre, F-67000 Strasbourg, France
³ Department of Medicine II, University of Freiburg, Hugstetter Straße 55, D-79106 Freiburg, Germany
⁴ Instituto Butantan, Laboratório de Imunologia Viral, 05503-900 São Paulo, Brazil

Correspondence
Jean-Pierre Martin
jp.martin{at}viro-ulp.u-strasbg.fr

Hepatitis C virus (HCV) is a major cause of chronic hepatitis worldwide. Studies of the early steps of HCV infection have been hampered by the lack of convenient in vitro or in vivo models. Although several cell-surface molecules that mediate the binding of HCV envelope proteins to target cells have been identified, mechanisms of viral entry into human hepatocytes are still poorly understood. Vesicular stomatitis virus/HCV pseudotyped viruses expressing the HCV envelope glycoproteins on the viral envelope were generated and it was found that their entry into human hepatocytes required co-expression of E1 and E2 on the pseudotype surface. Neutralization of pseudotype infection by anti-HCV antibodies suggested that cellular entry was mediated by HCV envelope glycoproteins and by previously characterized cell-surface molecules, including CD81. An entry assay based on the release of a fluorochrome from labelled HCV pseudotypes provided evidence for a pH-dependent fusion of the pseudotype envelope with a cellular compartment. By using a panel of endocytosis inhibitors, it is postulated that penetration of HCV into primary cultures of hepatocytes takes place by clathrin-mediated endocytosis.

Present address: Centre INRA de Clermont-Theix, Unité Nutrition et Métabolisme Protéique, 63122 Ceyrat, France.

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