Skip to main content
SpringerLink
Log in

Cell Culture-Based Assays to Test for Bacterial Adherence and Internalization

  • Protocol
  • First Online:
Helicobacter Species

Part of the book series: Methods in Molecular Biology ((MIMB,volume 921))

Abstract

Adherence and internalization of Helicobacter pylori into epithelial cells is a recently recognized event in the pathogen’s life cycle.

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 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

Notes

  1. 1.

    If a microaerophilic incubator is not available, evacuation anaerobic jars can be used to attain the necessary microaerophilic conditions by flushing a microaerophilic gaseous mixture into the sealed jars following evacuation. The sealed jars are then placed in a 37°C incubator for 72 h. Long-term storage of H. pylori can be achieved by adding 10% sterile glycerol to the Brucella bacterial culture broth, aliquoting the mixture into sterile 2 mL cryovials (Simport, T310-2A, Beloeil, QC), and storing at −70°C.

References

  1. Peek RM Jr, Blaser MJ (2002) Helicobacter pylori and gastrointestinal tract adenocarcinomas. Nat Rev Cancer 2:28–37

    Article  PubMed  CAS  Google Scholar 

  2. Peek RM Jr, Crabtree JE (2006) Helicobacter infection and gastric neoplasia. J Pathol 208:233–248

    Article  PubMed  CAS  Google Scholar 

  3. Amieva MR, Salama NR, Tompkins LS, Falkow S (2002) Helicobacter pylori enter and survive within multivesicular vacuoles of epithelial cells. Cell Microbiol 4:677–690

    Article  PubMed  CAS  Google Scholar 

  4. Terebiznik MR et al (2006) Helicobacter pylori VacA toxin promotes bacterial intracellular survival in gastric epithelial cells. Infect Immun 74:6599–6614

    Article  PubMed  CAS  Google Scholar 

  5. Dubois A (1995) Spiral bacteria in the human stomach: the gastric helicobacters. Emerg Infect Dis 1:79–85

    Article  PubMed  CAS  Google Scholar 

  6. Wyle FA, Tarnawski A, Schulman D, Dabros W (1990) Evidence for gastric mucosal cell invasion by C. pylori: an ultrastructural study. J Clin Gastroenterol 12(Suppl 1): S92–98

    Article  PubMed  Google Scholar 

  7. Petersen AM, Blom J, Andersen LP, Krogfelt KA (2000) Role of strain type, AGS cells and fetal calf serum in Helicobacter pylori adhesion and invasion assays. FEMS Immunol Med Microbiol 29:59–67

    Article  PubMed  CAS  Google Scholar 

  8. Petersen AM, Krogfelt KA (2003) Helicobacter pylori: an invading microorganism? A review. FEMS Immunol Med Microbiol 36: 117–126

    Article  PubMed  CAS  Google Scholar 

  9. Petersen AM, Sorensen K, Blom J, Krogfelt KA (2001) Reduced intracellular survival of Helicobacter pylori vacA mutants in comparison with their wild-types indicates the role of VacA in pathogenesis. FEMS Immunol Med Microbiol 30:103–108

    Article  PubMed  CAS  Google Scholar 

  10. Rittig MG et al (2003) Helicobacter pylori-induced homotypic phagosome fusion in human monocytes is independent of the bacterial vacA and cag status. Cell Microbiol 5:887–899

    Article  PubMed  CAS  Google Scholar 

  11. Oh JD, Karam SM, Gordon JI (2005) Intracellular Helicobacter pylori in gastric epithelial progenitors. Proc Natl Acad Sci USA 102:5186–5191

    Article  PubMed  CAS  Google Scholar 

  12. Rieder G, Fischer W, Haas R (2005) Interaction of Helicobacter pylori with host cells: function of secreted and translocated molecules. Curr Opin Microbiol 8:67–73

    Article  PubMed  CAS  Google Scholar 

  13. Bronte-Tinkew DM et al (2009) Helicobacter pylori cytotoxin-associated gene A activates the signal transducer and activator of transcription 3 pathway in vitro and in vivo. Cancer Res 69:632–639

    Article  PubMed  CAS  Google Scholar 

  14. Terebiznik MR et al (2009) Effect of Helicobacter pylori’s vacuolating cytotoxin on the autophagy pathway in gastric epithelial cells. Autophagy 5:370–379

    Article  PubMed  CAS  Google Scholar 

  15. Bjorkholm B et al (2000) Helicobacter pylori entry into human gastric epithelial cells: a potential determinant of virulence, persistence, and treatment failures. Helicobacter 5:148–154

    Article  PubMed  CAS  Google Scholar 

  16. Correa P, Houghton J (2007) Carcinogenesis of Helicobacter pylori. Gastroenterology 133:659–672

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

NLJ is supported by a Canadian Institute for Health Research operating grant (No. 17886).

Author information

Authors and Affiliations

  1. Cell Biology Program, Research Institute, Hospital for Sick Children, Toronto, ON, Canada

    Deepa Raju, David Rizzuti & Nicola L. Jones

Authors
  1. Deepa Raju
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David Rizzuti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nicola L. Jones
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nicola L. Jones .

Editor information

Editors and Affiliations

  1. Medical School, Dept. Medicine, University of Massachusetts, Plantation Street 364, Worcester, 01635, Massachusetts, USA

    JeanMarie Houghton

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Raju, D., Rizzuti, D., Jones, N.L. (2012). Cell Culture-Based Assays to Test for Bacterial Adherence and Internalization. In: Houghton, J. (eds) Helicobacter Species. Methods in Molecular Biology, vol 921. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-005-2_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-005-2_10

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-004-5

  • Online ISBN: 978-1-62703-005-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation