Skip to main content

Viral Nucleic Acid Recognition in Human Nonimmune Cells: In Vitro Systems

  • Protocol
  • First Online:
Innate DNA and RNA Recognition

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1169))

  • 1668 Accesses

Abstract

The innate immune system detects viral infections through a variety of receptors that sense the virus via their nucleic acid genome and evokes initial antiviral responses. Nonimmune cells possess pathogen-recognition mechanisms that enable them to respond to virus by the expression of RNA or DNA recognition receptors, thereby initiating the first steps in the host-pathogen interaction and inducing the production of pro-inflammatory and antiviral mediators. To better understand the activation of innate-immune defense mechanisms by nonimmune cells, we describe an experimental procedure that mimics viral infection, using four human nonimmune cell culture models under stimulation with synthetic analogues of RNA and DNA virus. Furthermore, we exemplify two viral infection models using cultured nonimmune cells and hepatitis C virus (HCV) or polyomavirus BK (BKV). In addition, we report the importance of siRNA technique in gene regulation in order to identify specific markers involved in innate antiviral responses in these cells.

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 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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. Takeushi O, Akira S (2009) Innate immunity to virus infection. Immunol Rev 227:75–86

    Article  Google Scholar 

  2. Kawai T, Akira S (2009) The roles of TLRs, RLRs and NLRs in pathogen recognition. Int Immunol 21:317–337

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Hornung V, Ablasser A, Charrel-Dennis M, Bauernfeind F, Horvath G, Caffrey DR, Latz E, Fitzgerald KA (2009) AIM2 recognizes cytosolic dsDNA and forms a caspase-1 activating inflammasome with ASC. Nature 458:514–518

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Akira S, Uematsu S, Takeuchi O (2006) Pathogen recognition and innate immunity. Cell 124:783–801

    Article  CAS  PubMed  Google Scholar 

  5. Abreu MT, Fukata M, Arditi M (2005) TLR signaling in the gut in health and disease. J Immunol 174:4453–4460

    Article  CAS  PubMed  Google Scholar 

  6. Fritz JH, Le Bourhis L, Magalhaes JG, Philpott DJ (2008) Innate immune recognition at the epithelial barrier drives adaptive immunity: APCs take the back seat. Trends Immunol 29:41–49

    Article  CAS  PubMed  Google Scholar 

  7. Ueta M, Hamuro J, Kiyono H, Kinoshita S (2005) Triggering of TLR3 by polyI:C in human corneal epithelial cells to induce inflammatory cytokines. Biochem Biophys Res Commun 331:285–294

    Article  CAS  PubMed  Google Scholar 

  8. Guillot L, Le Goffic R, Bloch S, Escriou N, Akira S, Chignard M, Si-Tahar M (2005) Involvement of toll-like receptor 3 in the immune response of lung epithelial cells to double-stranded RNA and influenza A virus. J Biol Chem 280:5571–5580

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Andrea Ribeiro .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, New York

About this protocol

Cite this protocol

Ribeiro, A., Wörnle, M. (2014). Viral Nucleic Acid Recognition in Human Nonimmune Cells: In Vitro Systems. In: Anders, HJ., Migliorini, A. (eds) Innate DNA and RNA Recognition. Methods in Molecular Biology, vol 1169. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0882-0_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0882-0_7

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0881-3

  • Online ISBN: 978-1-4939-0882-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics