Entry of hepatitis C virus pseudotypes into primary human hepatocytes by clathrin-dependent endocytosis.
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.
Codran
Audrey
A
INSERM U544, EA 3770, Institut de Virologie, Université Louis Pasteur, 3 rue Koeberlé, F-67000 Strasbourg, France.
Royer
Cathy
C
INSERM U544, EA 3770, Institut de Virologie, Université Louis Pasteur, 3 rue Koeberlé, F-67000 Strasbourg, France.
Jaeck
Daniel
D
Centre de Chirurgie Viscérale et de Transplantation, Centre Hospitalier de Hautepierre, F-67000 Strasbourg, France.
Bastien-Valle
Michèle
M
INSERM U544, EA 3770, Institut de Virologie, Université Louis Pasteur, 3 rue Koeberlé, F-67000 Strasbourg, France.
Baumert
Thomas F
TF
Department of Medicine II, University of Freiburg, Hugstetter Straße 55, D-79106 Freiburg, Germany.
Kieny
Marie Paule
MP
INSERM U544, EA 3770, Institut de Virologie, Université Louis Pasteur, 3 rue Koeberlé, F-67000 Strasbourg, France.
Pereira
Carlos Augusto
CA
Instituto Butantan, Laboratório de Imunologia Viral, 05503-900 São Paulo, Brazil.
Martin
Jean-Pierre
JP
INSERM U544, EA 3770, Institut de Virologie, Université Louis Pasteur, 3 rue Koeberlé, F-67000 Strasbourg, France.
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000821:virology; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002966:Clathrin / Q000502:physiology*; D006224:Cricetinae; D004279:DNA, Viral / Q000235:genetics; D004705:Endocytosis; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D022781:Hepatocytes / Q000502:physiology / Q000821:virology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D060225:Tetraspanin 28; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology
D015703:Antigens, CD; C554798:CD81 protein, human; D002966:Clathrin; D004279:DNA, Viral; C087035:E1 protein, Hepatitis C virus; D011993:Recombinant Fusion Proteins; D060225:Tetraspanin 28; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus