The Journal of Experimental Medicine
Fluorescence In Vivo Endomicroscopy
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Published 3 March 2003. doi:10.1084/jem.20021756
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© Rockefeller University Press, 0022-1007/2003/3/633 $5.00
The Journal of Experimental Medicine, Volume 197, Number 5, 633-642

Infectious Hepatitis C Virus Pseudo-particles Containing Functional E1–E2 Envelope Protein Complexes

Birke Bartosch1, Jean Dubuisson2 and François-Loïc Cosset1

1 Laboratoire de Vectorologie Rétrovirale et Thérapie Génique, Institut National de la Santé et de la Recherche Médicale U412, IFR 128, Ecole Normale Supérieure de Lyon, 69364 Lyon Cedex 07, France
2 Centre National de la Recherche Scientifique-UPR2511, Institut de Biologie de Lille, Institut Pasteur de Lille, 59021 Lille Cedex, France

Address correspondence to François-Loïc Cosset, LVRTG, ENS de Lyon, 46 Allée d'Italie, 69364 Lyon Cedex 07, France. Phone: 33-472-72-87-32; Fax: 33-472-72-80-80; E-mail: flcosset{at}ens-lyon.fr

The study of hepatitis C virus (HCV), a major cause of chronic liver disease, has been hampered by the lack of a cell culture system supporting its replication. Here, we have successfully generated infectious pseudo-particles that were assembled by displaying unmodified and functional HCV glycoproteins onto retroviral and lentiviral core particles. The presence of a green fluorescent protein marker gene packaged within these HCV pseudo-particles allowed reliable and fast determination of infectivity mediated by the HCV glycoproteins. Primary hepatocytes as well as hepato-carcinoma cells were found to be the major targets of infection in vitro. High infectivity of the pseudo-particles required both E1 and E2 HCV glycoproteins, and was neutralized by sera from HCV-infected patients and by some anti-E2 monoclonal antibodies. In addition, these pseudo-particles allowed investigation of the role of putative HCV receptors. Although our results tend to confirm their involvement, they provide evidence that neither LDLr nor CD81 is sufficient to mediate HCV cell entry. Altogether, these studies indicate that these pseudo-particles may mimic the early infection steps of parental HCV and will be suitable for the development of much needed new antiviral therapies.

Key Words: hepatitis • viral assembly • glycoproteins • receptor • neutralization


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