HBV infection of cell culture: evidence for multivalent and cooperative attachment

EMBO J. 2001 Aug 15;20(16):4443-53. doi: 10.1093/emboj/20.16.4443.


Hepadnaviruses do not infect cultured cells, therefore our knowledge of the mechanism of the early stages of virus-cell interaction is rather poor. In this study, we show that dimethylsulfoxide (DMSO)-treated HepG2 hepatoblastoma cells are infected efficiently by serum-derived hepatitis B virus (HBV) as monitored by viral gene expression and replication markers. To measure virus attachment, a variety of HBV surface proteins (HBsAgs) were conjugated to polystyrene beads and their capacity to attach cells was visualized and quantified by light microscopy at a single-cell resolution. Remarkably, DMSO increases the attachment efficiency by >200-fold. We further identify the QLDPAF sequence within preS1 as the receptor-binding viral domain epitope. Interestingly, a similar sequence is shared by several cellular, bacterial and viral proteins involved in cell adhesion, attachment and fusion. We also found that the small HBsAg contains a secondary attachment site that recognizes a distinct receptor on the cell membrane. Furthermore, we provide evidence in support of multivalent HBV attachment with synergistic interplay. Our data depict a mechanistic view of virus attachment and ingestion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Cell Culture Techniques
  • Detergents / pharmacology
  • Dimethyl Sulfoxide / pharmacology
  • Endocytosis
  • Hepatitis B Surface Antigens / genetics
  • Hepatitis B Surface Antigens / metabolism*
  • Hepatitis B virus / genetics
  • Hepatitis B virus / metabolism*
  • Humans
  • Membrane Fusion / physiology
  • Molecular Sequence Data
  • Protein Precursors / genetics
  • Protein Precursors / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Tumor Cells, Cultured
  • Virion


  • Detergents
  • Hepatitis B Surface Antigens
  • Protein Precursors
  • Recombinant Fusion Proteins
  • presurface protein 1, hepatitis B surface antigen
  • Dimethyl Sulfoxide