Proteoglycans act as cellular hepatitis delta virus attachment receptors

PLoS One. 2013;8(3):e58340. doi: 10.1371/journal.pone.0058340. Epub 2013 Mar 7.

Abstract

The hepatitis delta virus (HDV) is a small, defective RNA virus that requires the presence of the hepatitis B virus (HBV) for its life cycle. Worldwide more than 15 million people are co-infected with HBV and HDV. Although much effort has been made, the early steps of the HBV/HDV entry process, including hepatocyte attachment and receptor interaction are still not fully understood. Numerous possible cellular HBV/HDV binding partners have been described over the last years; however, so far only heparan sulfate proteoglycans have been functionally confirmed as cell-associated HBV attachment factors. Recently, it has been suggested that ionotrophic purinergic receptors (P2XR) participate as receptors in HBV/HDV entry. Using the HBV/HDV susceptible HepaRG cell line and primary human hepatocytes (PHH), we here demonstrate that HDV entry into hepatocytes depends on the interaction with the glycosaminoglycan (GAG) side chains of cellular heparan sulfate proteoglycans. We furthermore provide evidence that P2XR are not involved in HBV/HDV entry and that effects observed with inhibitors for these receptors are a consequence of their negative charge. HDV infection was abrogated by soluble GAGs and other highly sulfated compounds. Enzymatic removal of defined carbohydrate structures from the cell surface using heparinase III or the obstruction of GAG synthesis by sodium chlorate inhibited HDV infection of HepaRG cells. Highly sulfated P2XR antagonists blocked HBV/HDV infection of HepaRG cells and PHH. In contrast, no effect on HBV/HDV infection was found when uncharged P2XR antagonists or agonists were applied. In summary, HDV infection, comparable to HBV infection, requires binding to the carbohydrate side chains of hepatocyte-associated heparan sulfate proteoglycans as attachment receptors, while P2XR are not actively involved.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Glycosaminoglycans / chemistry
  • Glycosaminoglycans / metabolism
  • Hepatitis Delta Virus / physiology*
  • Humans
  • Proteoglycans / chemistry
  • Proteoglycans / metabolism*
  • Receptors, Virus / metabolism*
  • Static Electricity
  • Sulfates / chemistry
  • Virus Attachment*

Substances

  • Glycosaminoglycans
  • Proteoglycans
  • Receptors, Virus
  • Sulfates

Grants and funding

Oscar Lamas was supported by a Sheila Dame Postdoctoral Grant awarded by the European Association for the Study of the Liver (EASL) (www.easl.eu). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.