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Published on: October 22, 2013
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Clostridium difficile-mediated effects on human intestinal epithelia: Modelling host-pathogen interactions in a
Nazila V Jafari1, Sarah A Kuehne2, Nigel P Minton2
1Infection, Immunity, Inflammation and Physiological Medicine, Institute of Child Health, University College London, London, UK.
Anaerobe
|December 29, 2015
Summary
Clostridium difficile infection involves complex interactions between the bacterium and human intestinal cells. Anaerobic conditions significantly impact this crosstalk, influencing immune responses and epithelial barrier function.
Area of Science:
- Microbiology
- Immunology
- Gastroenterology
Background:
- Clostridium difficile infection (CDI) is a major cause of healthcare-associated diarrhea.
- While C. difficile toxins are understood, host-pathogen interactions in the gut remain unclear.
- The role of anaerobic environments in CDI pathogenesis needs further investigation.
Purpose of the Study:
- To investigate C. difficile strains' interactions with human intestinal epithelial cells (IECs).
- To determine if C. difficile mediates IEC antimicrobial immunity through toxin-dependent and independent mechanisms.
- To assess the impact of anaerobic conditions on bacterial-IEC interactions.
Main Methods:
- Co-culture of C. difficile PCR ribotypes (RT027, RT012, RT017) with IEC cell lines (HEK, Caco-2, T84).
- Measurement of IL-8 release from TLR-transfected HEK cells to assess immune engagement.
- Ussing chamber system to evaluate crosstalk under anaerobic versus standard CO2 conditions, measuring cytokine release and trans-epithelial electrical resistance (TEER).
Main Results:
- All C. difficile RTs engaged with human Toll-like receptors (TLR)-5, TLR2-CD14, and TLR2/6.
- Toxins TcdA and TcdB had minimal impact on bacterial adherence to Caco-2 cells.
- Anaerobic conditions in an Ussing chamber significantly increased C. difficile-T84 crosstalk, cytokine release, and TEER dysfunction compared to 5% CO2.
Conclusions:
- C. difficile engages with human IECs via TLRs.
- Bacterial adherence is minimally affected by toxins but influenced by anaerobic conditions.
- Anaerobic C. difficile interaction with human IECs is a complex process involving bacterial and toxin-mediated events impacting immunity and barrier function.
