Skip to main content

Human Papillomavirus Type 31 Life Cycle Methods for Study Using Tissue Culture Models

  • Protocol
DNA Viruses

Part of the book series: Methods in Molecular Biology ((MIMB,volume 292))

Abstract

The life cycle of human papillomaviruses (HPVs) has been difficult to study in tissue culture owing to its dependence on epithelial differentiation. In this chapter several methods are described to imitate the important steps in the HPV life cycle. Normal human keratinocytes (NHKs) harvested from neonatal foreskins were transfected with HPV type 31 genomes in order to generate stable cell lines containing episomal copies of HPV genomes. HPV-positive keratinocyte cultures were maintained in E medium in the presence of mitomycin C-treated J2 3T3 fibroblast feeders. Finally, the keratinocytes were induced to undergo epithelial differentiation in semisolid medium to provoke viral late functions like genomic amplification and late transcription.

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 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
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. Howley, P. M. and Lowy, D. R. (2001) Papillomaviruses and their replication, in Virology, vol. 2 (Fields, B. N., Knipe, J. B., and Howley, P. M., eds.), Lippincott Williams & Wilkins, Philadelphia, pp. 2197–2229.

    Google Scholar 

  2. Stubenrauch, F. and Laimins, L. A. (1999) Human papillomavirus life cycle: active and latent phases. Semin. Cancer Biol. 9, 379–386.

    Article  PubMed  CAS  Google Scholar 

  3. Lambert, P. F. (1991) Papillomavirus DNA replication. J. Virol. 65, 3417–3420.

    PubMed  CAS  Google Scholar 

  4. Laimins, L. A. (1996) Human papillomaviruses target differentiating epithelia for virion production and malignant conversion. Semin. Virol. 7, 305–313.

    Article  CAS  Google Scholar 

  5. Meyers, C., Mayer, T. J., and Ozbun, M. A. (1997) Synthesis of infectious human papillomavirus type 18 in differentiating epithelium transfected with viral DNA. J. Virol. 71, 7381–7386.

    PubMed  CAS  Google Scholar 

  6. Frattini, M. G., Lim, H. B., Doorbar, J., and Laimins, L. A. (1997) Induction of human papillomavirus type 18 late gene expression and genomic amplification in organotypic cultures from transfected DNA templates. J. Virol. 71, 7068–7072.

    PubMed  CAS  Google Scholar 

  7. Frattini, M. G., Lim, H. B., and Laimins, L. A. (1996) In vitro synthesis of oncogenic human papillomaviruses requires episomal genomes for differentiationdependent late expression. Proc. Natl. Acad. Sci. USA 93, 3062–3067.

    Article  PubMed  CAS  Google Scholar 

  8. McCance, D. J., Kopan, R., Fuchs, E., and Laimins, L. A. (1988) Human papillomavirus type 16 alters human epithelial cell differentiation in vitro. Proc. Natl. Acad. Sci. USA 85, 7169–7173.

    Article  PubMed  CAS  Google Scholar 

  9. Meyers, C. and Laimins, L. A. (1994) In vitro systems for the study and propagation of human papillomaviruses. Curr. Top. Microbiol. Immunol. 186, 199–215.

    PubMed  CAS  Google Scholar 

  10. Flores, E. R. and Lambert, P. F. (1997) Evidence for a switch in the mode of human papillomavirus type 16 DNA replication during the viral life cycle. J. Virol. 71, 7167–7179.

    PubMed  CAS  Google Scholar 

  11. Green, H. (1977) Terminal differentiation of cultured human epidermal cells. Cell 11, 405–416.

    Article  PubMed  CAS  Google Scholar 

  12. Ruesch, M. N. and Laimins, L. A. (1998) Human papillomavirus oncoproteins alter differentiation-dependent cell cycle exit on suspension in semisolid medium. Virology 250, 19–29.

    Article  PubMed  CAS  Google Scholar 

  13. Ruesch, M. N., Stubenrauch, F. and Laimins, L. A. (1998) Activation of papillomavirus late gene transcription and genome amplification upon differentiation in semisolid medium is coincident with expression of involucrin and transglutaminase but not keratin-10. J. Virol. 72, 5016–5024.

    PubMed  CAS  Google Scholar 

  14. Fehrmann, F., Klumpp, D. J. and Laimins, L. A. (2003) Human papillomavirus type 31 E5 protein supports cell cycle progression and activates late viral functions upon epithelial differentiation. J. Virol. 77, 2819–2831.

    Article  PubMed  CAS  Google Scholar 

  15. Thomas, J. T., Oh, S. T., Terhune, S. S., and Laimins, L. A. (2001) Cellular changes induced by low-risk human papillomavirus type 11 in keratinocytes that stably maintain viral episomes. J. Virol. 75, 7564–7571.

    Article  PubMed  CAS  Google Scholar 

  16. Terhune, S. S., Hubert, W. G., Thomas, J. T., and Laimins, L. A. (2001) Early polyadenylation signals of human papillomavirus type 31 negatively regulate capsid gene expression. J. Virol. 75, 8147–8157.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Humana Press Inc.

About this protocol

Cite this protocol

Fehrmann, F., Laimins, L.A. (2005). Human Papillomavirus Type 31 Life Cycle Methods for Study Using Tissue Culture Models. In: Lieberman, P.M. (eds) DNA Viruses. Methods in Molecular Biology, vol 292. Humana Press. https://doi.org/10.1385/1-59259-848-X:317

Download citation

  • DOI: https://doi.org/10.1385/1-59259-848-X:317

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-353-4

  • Online ISBN: 978-1-59259-848-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics