Skip to main content
SpringerLink
Log in

The Use of Hepatocytes to Investigate HDV Infection: The HDV/HepaRG Model

  • Protocol
  • First Online:
Hepatocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 640))

Abstract

Worldwide, it is estimated that more than 350 million people are chronically infected with hepatitis B virus (HBV), approximately 15 million of whom are coinfected with hepatitis D virus (HDV), a satellite of HBV that uses the envelope proteins of the latter to assemble its infectious particles. For a long time after HBV discovery, research on the viral life cycle, viral entry in particular, has been hampered by the lack of practical tissue culture systems. To date, in vitro isolation and serial propagation of HBV are still problematic, but the examination of the entire HBV life cycle is possible using two separate systems: (i) permissive human hepatoma cell lines to study HBV DNA replication, viral transcription, translation, assembly, and release of viral particles and (ii) primary cultures of human or chimpanzee hepatocytes or the susceptible HepaRG cell line for viral entry examination. The experimental model described here for analyzing the function of HBV envelope proteins at viral entry is based on this dual tissue culture system, in which HDV is substituted to HBV for practical reasons.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Seeger, C. and Mason, W.S. (2000) Hepatitis B virus biology. Microbiol. Mol. Biol. Rev. 64, 51–68.

    Article  PubMed  CAS  Google Scholar 

  2. Fournier, C., Sureau, C., Coste, J., Ducos, J., Pageaux, G., Larrey, D., Domergue, J., and Maurel, P. (1998) In vitro infection of adult normal human hepatocytes in primary culture by hepatitis C virus. J. Gen. Virol. 79 (Pt 10), 2367–2374.

    PubMed  CAS  Google Scholar 

  3. Lanford, R.E., Sureau, C., Jacob, J.R., White, R., and Fuerst, T.R. (1994) Demonstration of in vitro infection of chimpanzee hepatocytes with hepatitis C virus using strand-specific RT/PCR. Virology 202, 606–614.

    Article  PubMed  CAS  Google Scholar 

  4. Sureau, C. (1993) In vitro culture systems for hepatitis B and delta viruses. Arch. Virol. Suppl. 8, 3–14.

    PubMed  CAS  Google Scholar 

  5. Sureau, C., Romet-Lemonne, J.L., Mullins, J.I., and Essex, M. (1986) Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA. Cell 47, 37–47.

    Article  PubMed  CAS  Google Scholar 

  6. Chang, C.M., Jeng, K.S., Hu, C.P., Lo, S.J., Su, T.S., Ting, L.P., Chou, C.K., Han, S.H., Pfaff, E., Salfeld, J. et al. (1987) Production of hepatitis B virus in vitro by transient expression of cloned HBV DNA in a hepatoma cell line. EMBO J. 6, 675–680.

    PubMed  CAS  Google Scholar 

  7. Sells, M.A., Chen, M.L., and Acs, G. (1987) Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc. Natl. Acad. Sci. USA 84, 1005–1009.

    Article  PubMed  CAS  Google Scholar 

  8. Tsurimoto, T., Fujiyama, A., and Matsubara, K. (1987) Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. Proc. Natl. Acad. Sci. USA 84, 444–448.

    Article  PubMed  CAS  Google Scholar 

  9. Yaginuma, K., Shirakata, Y., Kobayashi, M., and Koike, K. (1987) Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA. Proc. Natl. Acad. Sci. USA 84, 2678–2682.

    Article  PubMed  CAS  Google Scholar 

  10. Jacob, J.R., Eichberg, J.W., and Lanford, R.E. (1989) In vitro replication and expression of hepatitis B virus from chronically infected primary chimpanzee hepatocytes. Hepatology 10, 921–927.

    Article  PubMed  CAS  Google Scholar 

  11. Gripon, P., Diot, C., and Guguen-Guillouzo, C. (1993) Reproducible high level infection of cultured adult human hepatocytes by hepatitis B virus: effect of polyethylene glycol on adsorption and penetration. Virology 192, 534–540.

    Article  PubMed  CAS  Google Scholar 

  12. Gripon, P., Diot, C., Theze, N., Fourel, I., Loreal, O., Brechot, C., and Guguen-Guillouzo, C. (1988) Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. J. Virol. 62, 4136–4143.

    PubMed  CAS  Google Scholar 

  13. Sureau, C., Jacob, J.R., Eichberg, J.W., and Lanford, R.E. (1991) Tissue culture system for infection with human hepatitis delta virus. J. Virol. 65, 3443–3450.

    PubMed  CAS  Google Scholar 

  14. Blanc, P., Desprez, D., Fabre, J.M., Pageaux, G., Daures, J.P., Larrey, D., Saint-Aubert, B., Michel, H., and Maurel, P. (1996) Contribution of primary cultures of adult human hepatocytes to the pathophysiology of hepatocellular carcinoma. J. Hepatol. 25, 663–669.

    Article  PubMed  CAS  Google Scholar 

  15. Ferrini, J.B., Ourlin, J.C., Pichard, L., Fabre, G., and Maurel, P. (1998) Human hepatocyte culture. Methods Mol. Biol. 107, 341–352.

    PubMed  CAS  Google Scholar 

  16. Pichard-Garcia, L., Gerbal-Chaloin, S., Ferrini, J.B., Fabre, J.M., and Maurel, P. (2002) Use of long-term cultures of human hepatocytes to study cytochrome P450 gene expression. Methods Enzymol. 357, 311–321.

    Article  PubMed  CAS  Google Scholar 

  17. Jacob, J.R., Burk, K.H., Eichberg, J.W., Dreesman, G.R., and Lanford, R.E. (1990) Expression of infectious viral particles by primary chimpanzee hepatocytes isolated during the acute phase of non-A, non-B hepatitis. J. Infect. Dis. 161, 1121–1127.

    Article  PubMed  CAS  Google Scholar 

  18. Gripon, P., Rumin, S., Urban, S., Le Seyec, J., Glaise, D., Cannie, I., Guyomard, C., Lucas, J., Trepo, C., and Guguen-Guillouzo, C. (2002) Infection of a human hepatoma cell line by hepatitis B virus. Proc. Natl. Acad. Sci. USA 99, 15655–15660.

    Article  PubMed  CAS  Google Scholar 

  19. Abou-Jaoude, G. and Sureau, C. (2007) Entry of hepatitis delta virus requires the conserved cysteine residues of the hepatitis B virus envelope protein antigenic loop and is blocked by inhibitors of thiol-disulfide exchange. J. Virol. 81, 13057–13066.

    Article  PubMed  CAS  Google Scholar 

  20. Blanchet, M. and Sureau, C. (2006) Analysis of the cytosolic domains of the hepatitis B virus envelope proteins for their function in viral particle assembly and infectivity. J. Virol. 80, 11935–11945.

    Article  PubMed  CAS  Google Scholar 

  21. Engelke, M., Mills, K., Seitz, S., Simon, P., Gripon, P., Schnolzer, M., and Urban, S. (2006) Characterization of a hepatitis B and hepatitis delta virus receptor binding site. Hepatology 43, 750–760.

    Article  PubMed  CAS  Google Scholar 

  22. Gripon, P., Cannie, I., and Urban, S. (2005) Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein. J. Virol. 79, 1613–1622.

    Article  PubMed  CAS  Google Scholar 

  23. Lucifora, J., Durantel, D., Belloni, L., Barraud, L., Villet, S., Vincent, I.E., Margeridon-Thermet, S., Hantz, O., Kay, A., Levrero, M., and Zoulim, F. (2008) Initiation of hepatitis B virus genome replication and production of infectious virus following delivery in HepG2 cells by novel recombinant baculovirus vector. J. Gen. Virol. 89, 1819–1828.

    Article  PubMed  CAS  Google Scholar 

  24. Schulze, A., Gripon, P., and Urban, S. (2007) Hepatitis B virus infection initiates with a large surface protein-dependent binding to heparan sulfate proteoglycans. Hepatology 46, 1759–1768.

    Article  PubMed  CAS  Google Scholar 

  25. Ganem, D. and Prince, A.M. (2004) Hepatitis B virus infection-natural history and clinical consequences. N. Engl. J. Med. 350, 1118–1129.

    Article  PubMed  CAS  Google Scholar 

  26. Ganem, D. and Schneider, R.J. (2001) Hepadnaviridae: the viruses and their replication, in Fields Virology (Knipe, D.M. and Howley, P.M. eds.), Lippincott Williams & Wilkins, Philadelphia PA USA, pp. 2923–2970.

    Google Scholar 

  27. Heermann, K.H. and Gerlich, W.H. (1992) Surface proteins of hepatitis B viruses, in Molecular Biology of HBV (Maclachlan, A. ed.), CRC Press, Boca Raton Florida USA.

    Google Scholar 

  28. Bonino, F., Heermann, K.H., Rizzetto, M., and Gerlich, W.H. (1986) Hepatitis delta virus: protein composition of delta antigen and its hepatitis B virus-derived envelope. J. Virol. 58, 945–950.

    PubMed  CAS  Google Scholar 

  29. Gerlich, W.H., Heermann, K.H., Ponzetto, A., Crivelli, O., and Bonino, F. (1987) Proteins of hepatitis delta virus. Prog. Clin. Biol. Res. 234, 97–103.

    PubMed  CAS  Google Scholar 

  30. Lai, M.M. (1995) The molecular biology of hepatitis delta virus. Annu. Rev. Biochem. 64, 259–286.

    Article  PubMed  CAS  Google Scholar 

  31. Taylor, J.M. (2006) Structure and replication of hepatitis delta virus RNA. Curr. Top. Microbiol. Immunol. 307, 1–23.

    Article  PubMed  CAS  Google Scholar 

  32. Taylor, J.M. (2006) Hepatitis delta virus. Virology 344, 71–76.

    Article  PubMed  CAS  Google Scholar 

  33. Sureau, C., Guerra, B., and Lanford, R.E. (1993) Role of the large hepatitis B virus envelope protein in infectivity of the hepatitis delta virion. J. Virol. 67, 366–372.

    PubMed  CAS  Google Scholar 

  34. Sureau, C. and Lanford, R. (1993) Analysis of hepatitis B virus envelope proteins in assembly and infectivity of human hepatitis delta virus. Prog. Clin. Biol. Res. 382, 45–51.

    PubMed  CAS  Google Scholar 

  35. Wang, C.J., Chen, P.J., Wu, J.C., Patel, D., and Chen, D.S. (1991) Small-form hepatitis B surface antigen is sufficient to help in the assembly of hepatitis delta virus-like particles. J. Virol. 65, 6630–6636.

    PubMed  CAS  Google Scholar 

  36. Barrera, A., Guerra, B., Lee, H., and Lanford, R.E. (2004) Analysis of host range phenotypes of primate hepadnaviruses by in vitro infections of hepatitis D virus pseudotypes. J. Virol. 78, 5233–5243.

    Article  PubMed  CAS  Google Scholar 

  37. Barrera, A., Guerra, B., Notvall, L., and Lanford, R.E. (2005) Mapping of the hepatitis B virus pre-S1 domain involved in receptor recognition. J. Virol. 79, 9786–9798.

    Article  PubMed  CAS  Google Scholar 

  38. Barrera, A. and Lanford, R.E. (2004) Infection of primary chimpanzee hepatocytes with recombinant hepatitis D virus particles: a surrogate model for hepatitis B virus. Methods Mol. Med. 96, 131–142.

    PubMed  Google Scholar 

  39. Blanchet, M. and Sureau, C. (2007) Infectivity determinants of the hepatitis B virus pre-S domain are confined to the N-terminal 75 amino acid residues. J. Virol. 81, 5841–5849.

    Article  PubMed  CAS  Google Scholar 

  40. Kuo, M.Y., Chao, M., and Taylor, J. (1989) Initiation of replication of the human hepatitis delta virus genome from cloned DNA: role of delta antigen. J. Virol. 63, 1945–1950.

    PubMed  CAS  Google Scholar 

  41. Sureau, C., Guerra, B., and Lee, H. (1994) The middle hepatitis B virus envelope protein is not necessary for infectivity of hepatitis delta virus. J. Virol. 68, 4063–4066.

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The author acknowledges O. Hantz and C. Trépo for the gift of the HepaRG cell line. CS is a CNRS investigator; he is supported by contracts with ANRS and INTS.

Author information

Authors and Affiliations

  1. Laboratoire de Virologie Moléculaire, Institut National de la Transfusion Sanguine, Paris, France

    Camille Sureau

Authors
  1. Camille Sureau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Camille Sureau .

Editor information

Editors and Affiliations

  1. INSERM U632, Labo. Physiopathologie Hépatique, Université Montpellier I, rte. Mende 1919, Montpellier, 34293, France

    Patrick Maurel

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Sureau, C. (2010). The Use of Hepatocytes to Investigate HDV Infection: The HDV/HepaRG Model. In: Maurel, P. (eds) Hepatocytes. Methods in Molecular Biology, vol 640. Humana Press. https://doi.org/10.1007/978-1-60761-688-7_25

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-688-7_25

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-687-0

  • Online ISBN: 978-1-60761-688-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation