Journal of Gastroenterology

, Volume 52, Issue 4, pp 452–465

A human gut ecosystem protects against C. difficile disease by targeting TcdA

  • Sarah Lynn Martz
  • Mabel Guzman-Rodriguez
  • Shu-Mei He
  • Curtis Noordhof
  • David John Hurlbut
  • Gregory Brian Gloor
  • Christian Carlucci
  • Scott Weese
  • Emma Allen-Vercoe
  • Jun Sun
  • Erika Chiong Claud
  • Elaine Olga Petrof
Original Article—Alimentary Tract

Abstract

Background

A defined Microbial Ecosystem Therapeutic (MET-1, or “RePOOPulate”) derived from the feces of a healthy volunteer can cure recurrent C. difficile infection (rCDI) in humans. The mechanisms of action whereby healthy microbiota protect against rCDI remain unclear. Since C. difficile toxins are largely responsible for the disease pathology of CDI, we hypothesized that MET-1 exerts its protective effects by inhibiting the effects of these toxins on the host.

Methods

A combination of in vivo (antibiotic-associated mouse model of C. difficile colitis, mouse ileal loop model) and in vitro models (FITC-phalloidin staining, F actin Western blots and apoptosis assay in Caco2 cells, transepithelial electrical resistance measurements in T84 cells) were employed.

Results

MET-1 decreased both local and systemic inflammation in infection and decreased both the cytotoxicity and the amount of TcdA detected in stool, without an effect on C.difficile viability. MET-1 protected against TcdA-mediated damage in a murine ileal loop model. MET-1 protected the integrity of the cytoskeleton in cells treated with purified TcdA, as indicated by FITC-phalloidin staining, F:G actin assays and preservation of transepithelial electrical resistance. Finally, co-incubation of MET-1 with purified TcdA resulted in decreased detectable TcdA by Western blot analysis.

Conclusions

MET-1 intestinal microbiota confers protection against C. difficile and decreases C. difficile-mediated inflammation through its protective effects against C. difficile toxins, including enhancement of host barrier function and degradation of TcdA. The effect of MET-1 on C. difficile viability seems to offer little, if any, contribution to its protective effects on the host.

Keywords

Microbiota C. difficile TcdA 

Supplementary material

535_2016_1232_MOESM1_ESM.pdf (571 kb)
Supplementary material 1 (PDF 570 kb)
535_2016_1232_MOESM2_ESM.pdf (68 kb)
Supplementary material 2 (PDF 68 kb)
535_2016_1232_MOESM3_ESM.pdf (7.8 mb)
Supplementary material 3 (PDF 8016 kb)

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Copyright information

© Japanese Society of Gastroenterology 2016

Authors and Affiliations

  • Sarah Lynn Martz
    • 1
  • Mabel Guzman-Rodriguez
    • 1
  • Shu-Mei He
    • 1
  • Curtis Noordhof
    • 1
  • David John Hurlbut
    • 2
  • Gregory Brian Gloor
    • 3
  • Christian Carlucci
    • 4
  • Scott Weese
    • 4
  • Emma Allen-Vercoe
    • 4
  • Jun Sun
    • 5
  • Erika Chiong Claud
    • 6
  • Elaine Olga Petrof
    • 1
  1. 1.Division of Infectious Diseases/GI Diseases Research Unit Wing, Department of Medicine, Kingston General HospitalQueen’s UniversityKingstonCanada
  2. 2.Department of Pathology and Molecular MedicineQueen’s UniversityKingstonCanada
  3. 3.Department of BiochemistryUniversity of Western OntarioLondonCanada
  4. 4.Department of Molecular and Cellular BiologyUniversity of GuelphGuelphCanada
  5. 5.Division of Gastroenterology and Hepatology, Department of MedicineUniversity of Illinois at ChicagoChicagoUSA
  6. 6.Departments of Pediatrics and MedicineUniversity of ChicagoChicagoUSA

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