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HBV infection of cell culture: evidence for multivalent and cooperative attachment
Author(s) -
Paran Nir,
Geiger Benjamin,
Shaul Yosef
Publication year - 2001
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/20.16.4443
Subject(s) - biology , hepatitis b virus , epitope , hbsag , virology , virus , viral entry , cell , cell culture , microbiology and biotechnology , viral replication , antibody , biochemistry , immunology , genetics
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.