Skip to main content

Enhancement of Infection of HepG2 Cells in Culture by Predigestion of Hepadnavirus with V8 Protease

  • Protocol
Hepatitis B and D Protocols

Part of the book series: Methods in Molecular Medicineā„¢ ((MIMM,volume 96))

  • 549 Accesses

Abstract

In this chapter, a method for infecting HepG2 cells with human hepatitis B virus (HBV) and woodchuck hepatitis virus (WHV) is reviewed. This chapter provides a step-wise, experimentally sensitive description of how to perform the protease infectivity assay. The assay is highly technical and sensitive to certain cell culture and virological steps. Therefore, essential technical points are emphasized, to assist the experimentalist as much as possible.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Lu, X., Block, T., and Gerlich, W. H. (1996) Protease-induced infectivity of hepatitis B virus for a human hepatoma cell line. J. Virol. 70, 2277ā€“2285.

    PubMedĀ  CASĀ  Google ScholarĀ 

  2. Lu, X., Hazboun, T. and Block, T.M. (2001) Limited proteolysis induces woodchuck hepatitis virus infectivity for human HepG2 cells. Virus Res. 73, 27ā€“40.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  3. Qiao, M., MacNaughton, T.B., and Gowans, E.J. (1994) Adsorption and penetration of hepatitis B virus in a nonpermissive cell line. Virology 201, 356ā€“363.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  4. Sells, M. A., Chen, M.L., and Acs, G. (1987) Production of hepatitis B virus particles in HepG2 cells transfected with cloned hepatitis B virus DNA. Proc. Natl. Acad. Sci. USA 84, 1005ā€“1009.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  5. Gripon, P., Diot, C., Theze, N., et al. (1988) Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. J. Virol. 62, 4136ā€“4143.

    PubMedĀ  CASĀ  Google ScholarĀ 

  6. 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, 534ā€“540.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  7. Ochiya, T., Tsurimoto, T., Ueda, K., Okubo, K., Shiozawa, M., and Matsubara, K. (1989) An in vitro system for infection with hepatitis B virus that uses primary human fetal hepatocytes. Proc Natl. Acad. Sci. USA 86, 1875ā€“1879.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  8. Galle, P.R., Haglestein, J., and Kommerell, B. (1994) In vitro experimental infection of primary human heptocytes with hepatitis B virus. Gastroenterology 106, 664ā€“673.

    PubMedĀ  CASĀ  Google ScholarĀ 

  9. Paran, N., Geiger, B., and Shaul, Y. (2001) HBV infection of cell culture: evidence for multivalent and cooperative attachment. EMBO J. 20, 4443ā€“4453.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  10. Li, J.S., Tong, S. P., and Wands, J. R. (1996) Characterization of a 120-kilodalton pre-S-binding protein as a candidate duck hepatitis B virus receptor. J. Viro. 70, 6029ā€“6035.

    CASĀ  Google ScholarĀ 

  11. Eng, F. J., Novikova, E. G., Kuroki, K., Ganem, D., and Fricker, L. D. (1998) gp180, a protein that binds duck hepatitis B virus particles, has metallocarboxypeptidase D-like enzymatic activity. J. Biol. Chem. 273, 8382ā€“8388.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  12. Qiao, M., Scougall, C.A., Duszynski, A., and Burrell1, C. J. (1999) Kinetics of early molecular events in duck hepatitis B virus replication in primary duck hepatocytes. J. Gen. Virol. 80, 2127ā€“2135.

    PubMedĀ  CASĀ  Google ScholarĀ 

  13. Gerlich, W.H., Lu, X., and Heermann, K.H. (1993) Studies on the attachment and penetration of hepatitis B virus. J. Hepatol. 17(Suppl. 3), S10ā€“S14.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  14. Oess, S. and Hildt, E. (2000) Novel cell permeable motif derived from the PreS2-domain of hepatitis-B virus surface antigens. Gene Ther. 7, 750ā€“758.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  15. Simsek, E., Ouzounov, S., Hazboum, T., Block, T.M. and Lu, X. A specific and sensitive method for quantification of HBV DNA. In preparation.

    Google ScholarĀ 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

Ā© 2004 Humana Press Inc.

About this protocol

Cite this protocol

Lu, X., Block, T.M. (2004). Enhancement of Infection of HepG2 Cells in Culture by Predigestion of Hepadnavirus with V8 Protease. In: Hamatake, R.K., Lau, J.Y.N. (eds) Hepatitis B and D Protocols. Methods in Molecular Medicineā„¢, vol 96. Humana Press. https://doi.org/10.1385/1-59259-670-3:199

Download citation

  • DOI: https://doi.org/10.1385/1-59259-670-3:199

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-108-0

  • Online ISBN: 978-1-59259-670-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics