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- Title
Identification and characterization of adhesive factors of <em>Clostridium difficile</em> involved in adhesion to human colonic enterocyte-like Caco-2 and mucus-secreting HT29 cells in culture.
- Authors
Eveillard, Matthieu; Fourel, Valérie; Barc, Marie-Claude; Kernéis, Sophie; Coconnier, Marie-Hélène; Karjalainen, Tuomo; Bourlioux, Pierre; Servin, Alain L.
- Abstract
Experiments reported in this communication showed that the highly toxinogenic Cd 79685, Cd 4784, and Wilkins <em>Clostridium difficile</em> strains and the moderately toxinogenic FD strain grown in the presence of blood adhere to polarized monolayers of two cultured human intestinal cell lines: the human colonic epithelial Caco-2 cells and the human mucus-secreting HT29-MTX cells. Scanning electron microscopy revealed that the bacteria interacted with well-defined apical microvilli of differentiated Caco-2 cells and that the bacteria strongly bind to the mucus layer that entirely covers the surface of the HT29-MTX cells. The binding of <em>C. difficile</em> to Caco-2 cells developed in parallel with the differentiation features of the Caco-2 cells, suggesting that the protein(s) which constitute <em>C. difficile</em>-binding sites are differentiation-related brush border protein(s). To better define this interaction, we tentatively characterized the mechanism(s) of adhesion of <em>C. difficile</em> with adherence assays. It was shown that heating of <em>C. difficile</em> grown in the presence of blood enhanced the bacterial interaction with the brush border of the enterocyte-like Caco-2 cells and the human mucus-secreting HT29-MTX cells. A labile surface-associated component was involved in <em>C. difficile</em> adhesion since washes of <em>C. difficile</em> grown in the presence of blood without heat shock decreased adhesion. After heating, washes of <em>C. difficile</em> grown in the presence of blood did not modify adhesion. Analysis of surface-associated proteins of <em>C. difficile</em> subjected to different culture conditions was conducted. After growth of <em>C. difficile</em> Cd 79685, Cd 4784, FD and Wilkins strains in the presence of blood and heating, two predominant SDS-extractable proteins with molecular masses of 12 and 27kDa were observed and two other proteins with masses of 48 and 31 kDa disappeared. Direct involvement of the 12 and 27kDa...
- Subjects
CLOSTRIDIOIDES difficile; CELLS; BACTERIA; PROTEINS; CLOSTRIDIUM
- Publication
Molecular Microbiology, 1993, Vol 7, Issue 3, p371
- ISSN
0950-382X
- Publication type
Article
- DOI
10.1111/j.1365-2958.1993.tb01129.x