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Journal of Virology, April 2007, p. 3608-3617, Vol. 81, No. 7
0022-538X/07/$08.00+0     doi:10.1128/JVI.02277-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Assembly of Hepatitis Delta Virus: Particle Characterization, Including the Ability To Infect Primary Human Hepatocytes{triangledown}

Severin Gudima,1 Yiping He,1 Anja Meier,1 Jinhong Chang,1 Rongji Chen,1 Michal Jarnik,1 Emmanuelle Nicolas,1 Volker Bruss,2 and John Taylor1*

Fox Chase Cancer Center, Philadelphia, Pennsylvania,1 Department of Virology, University of Göttingen, Göttingen, Germany2

Received 17 October 2006/ Accepted 8 January 2007

Efficient assembly of hepatitis delta virus (HDV) was achieved by cotransfection of Huh7 cells with two plasmids: one to provide expression of the large, middle, and small envelope proteins of hepatitis B virus (HBV), the natural helper of HDV, and another to initiate replication of the HDV RNA genome. HDV released into the media was assayed for HDV RNA and HBV envelope proteins and characterized by rate-zonal sedimentation, immunoaffinity purification, electron microscopy, and the ability to infect primary human hepatocytes. Among the novel findings were that (i) immunostaining for delta antigen 6 days after infection with 300 genome equivalents (GE) per cell showed only 1% of cells as infected, but this was increased to 16% when 5% polyethylene glycol was present during infection; (ii) uninfected cells did not differ from infected cells in terms of albumin accumulation or the presence of E-cadherin at cell junctions; and (iii) sensitive quantitative real-time PCR assays detected HDV replication even when the multiplicity of infection was 0.2 GE/cell. In the future, this HDV assembly and infection system can be further developed to better understand the mechanisms shared by HBV and HDV for attachment and entry into host cells.

* Corresponding author. Mailing address: Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111-2497. Phone: (215) 728-2436. Fax: (215) 728-3105. E-mail: john.taylor{at}fccc.edu.

{triangledown} Published ahead of print on 17 January 2007.

Journal of Virology, April 2007, p. 3608-3617, Vol. 81, No. 7
0022-538X/07/$08.00+0     doi:10.1128/JVI.02277-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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