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Journal of Virology, June 2001, p. 5084-5089, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.5084-5089.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Efficient Infection of Primary Tupaia Hepatocytes with Purified Human and Woolly Monkey Hepatitis B Virus

Josef Köck, Michael Nassal, Sabine MacNelly, Thomas F. Baumert, Hubert E. Blum, and Fritz von Weizsäcker*

Department of Medicine II, University of Freiburg, Freiburg, Germany

Received 17 November 2000/Accepted 6 March 2001

The Asian tree shrew, Tupaia belangeri, has been proposed as a novel animal model for studying hepatitis B virus (HBV) infection. Here, we describe a protocol for efficient and reproducible infection of primary tupaia hepatocytes with HBV. We report that human serum interferes with HBV binding to the hepatocytes, thus limiting the maximum multiplicity of infection. Purification of HBV virions by gradient sedimentation greatly enhances virus binding and infectivity. Covalently closed circular DNA was clearly detectable by Southern blot analysis and newly synthesized single-stranded HBV DNA was visible 2 weeks postinoculation. Primary tupaia hepatocytes are also susceptible to infection with the recently discovered woolly monkey hepatitis B virus (WMHBV) but not to woodchuck hepatitis virus infection. Compared to HBV, WMHBV replicated at a higher rate with single-stranded DNA detectable within the first week postinoculation. Primary tupaia hepatocytes should represent a useful system for studying early steps of HBV and WMHBV infection.

* Corresponding author. Mailing address: Department of Medicine II, University of Freiburg, Hugstetter Strasse 55, D-79106 Freiburg, Germany. Phone: 49-761-2703401. Fax: 49-761-2703610. E-mail: weiz{at}ukl.uni-freiburg.de.

Journal of Virology, June 2001, p. 5084-5089, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.5084-5089.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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