1887

Abstract

is a frequent cause of severe, recurrent, post-antibiotic diarrhoea and pseudomembranous colitis. Its pathogenicity is mediated mainly by two toxins, TcdA and TcdB. However, different adhesins have also been described as important colonization factors which are implicated in the first step of the intestinal infection. In this study, we focused our interest on one of these adhesins, fibronectin-binding protein A (FbpA), and on its role in the intestinal colonization process. A mutant of FbpA (CDΔFbpA) was constructed in strain 630Δerm by using ClosTron technology. This mutant was characterized and and compared to the isogenic wild-type strain. Adhesion of the CDΔFbpA mutant to the human colonic epithelial cell line Caco-2 and to mucus-secreting HT29-MTX cells was examined. Surprisingly, the CDΔFbpA mutant adhered more than the wild-type parental strain. The CDΔFbpA mutant was also analysed in three different mouse models by following the intestinal implantation kinetics (faecal shedding) and caecal colonization (7 days post-challenge). We showed that in monoxenic mice, CDΔFbpA shed in faeces at the same rate as that of the isogenic wild-type strain but its colonization of the caecal wall was significantly reduced. In dixenic mice, the shedding rate was slower for the CDΔFbpA mutant than for the isogenic wild-type strain during the first days of infection, but no significant difference was observed in caecal colonization. Similar rates of intestinal implantation and caecal colonization were observed for both strains in assays performed in human microbiota-associated mice. Taken together, our data suggest that FbpA plays a role in intestinal colonization by .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.029553-0
2011-08-01
2019-10-22
Loading full text...

Full text loading...

/deliver/fulltext/jmm/60/8/1155.html?itemId=/content/journal/jmm/10.1099/jmm.0.029553-0&mimeType=html&fmt=ahah

References

  1. Bellon-Fontaine M.-N., Rault J., van Oss C. J.. ( 1996;). Microbial adhesion to solvents: a novel method to determine the electron-donor/electron-acceptor or Lewis acid-base properties of microbial cells. . Colloids Surf B Biointerfaces 7:, 47–53. [CrossRef]
    [Google Scholar]
  2. Calabi E., Calabi F., Phillips A. D., Fairweather N. F.. ( 2002;). Binding of Clostridium difficile surface layer proteins to gastrointestinal tissues. . Infect Immun 70:, 5770–5778. [CrossRef].[PubMed]
    [Google Scholar]
  3. Cartman S. T., Heap J. T., Kuehne S. A., Cockayne A., Minton N. P.. ( 2010;). The emergence of ‘hypervirulence’ in Clostridium difficile. . Int J Med Microbiol 300:, 387–395. [CrossRef].[PubMed]
    [Google Scholar]
  4. Cerquetti M., Molinari A., Sebastianelli A., Diociaiuti M., Petruzzelli R., Capo C., Mastrantonio P.. ( 2000;). Characterization of surface layer proteins from different Clostridium difficile clinical isolates. . Microb Pathog 28:, 363–372. [CrossRef].[PubMed]
    [Google Scholar]
  5. Courtney H. S., Hasty D. L., Li Y., Chiang H. C., Thacker J. L., Dale J. B.. ( 1999;). Serum opacity factor is a major fibronectin-binding protein and a virulence determinant of M type 2 Streptococcus pyogenes. . Mol Microbiol 32:, 89–98. [CrossRef].[PubMed]
    [Google Scholar]
  6. Dramsi S., Bourdichon F., Cabanes D., Lecuit M., Fsihi H., Cossart P.. ( 2004;). FbpA, a novel multifunctional Listeria monocytogenes virulence factor. . Mol Microbiol 53:, 639–649. [CrossRef].[PubMed]
    [Google Scholar]
  7. Genth H., Dreger S. C., Huelsenbeck J., Just I.. ( 2008;). Clostridium difficile toxins: more than mere inhibitors of Rho proteins. . Int J Biochem Cell Biol 40:, 592–597. [CrossRef].[PubMed]
    [Google Scholar]
  8. Heap J. T., Pennington O. J., Cartman S. T., Carter G. P., Minton N. P.. ( 2007;). The ClosTron: a universal gene knock-out system for the genus Clostridium. . J Microbiol Methods 70:, 452–464. [CrossRef].[PubMed]
    [Google Scholar]
  9. Hennequin C., Porcheray F., Waligora-Dupriet A., Collignon A., Barc M., Bourlioux P., Karjalainen T.. ( 2001;). GroEL (Hsp60) of Clostridium difficile is involved in cell adherence. . Microbiology 147:, 87–96.[PubMed]
    [Google Scholar]
  10. Hennequin C., Janoir C., Barc M. C., Collignon A., Karjalainen T.. ( 2003;). Identification and characterization of a fibronectin-binding protein from Clostridium difficile. . Microbiology 149:, 2779–2787. [CrossRef].[PubMed]
    [Google Scholar]
  11. Holmes A. R., McNab R., Millsap K. W., Rohde M., Hammerschmidt S., Mawdsley J. L., Jenkinson H. F.. ( 2001;). The pavA gene of Streptococcus pneumoniae encodes a fibronectin-binding protein that is essential for virulence. . Mol Microbiol 41:, 1395–1408. [CrossRef].[PubMed]
    [Google Scholar]
  12. Janoir C., Péchiné S., Grosdidier C., Collignon A.. ( 2007;). Cwp84, a surface-associated protein of Clostridium difficile, is a cysteine protease with degrading activity on extracellular matrix proteins. . J Bacteriol 189:, 7174–7180. [CrossRef].[PubMed]
    [Google Scholar]
  13. Joh D., Wann E. R., Kreikemeyer B., Speziale P., Höök M.. ( 1999;). Role of fibronectin-binding MSCRAMMs in bacterial adherence and entry into mammalian cells. . Matrix Biol 18:, 211–223. [CrossRef].[PubMed]
    [Google Scholar]
  14. Lin Y. P., Kuo C. J., Koleci X., McDonough S. P., Chang Y. F.. ( 2011;). Manganese binds to Clostridium difficile Fbp68 and is essential for fibronectin binding. . J Biol Chem 286:, 3957–3969. [CrossRef].[PubMed]
    [Google Scholar]
  15. Molinari G., Talay S. R., Valentin-Weigand P., Rohde M., Chhatwal G. S.. ( 1997;). The fibronectin-binding protein of Streptococcus pyogenes, SfbI, is involved in the internalization of group A streptococci by epithelial cells. . Infect Immun 65:, 1357–1363.[PubMed]
    [Google Scholar]
  16. Muñoz-Provencio D., Pérez-Martínez G., Monedero V.. ( 2010;). Characterization of a fibronectin-binding protein from Lactobacillus casei BL23. . J Appl Microbiol 108:, 1050–1059. [CrossRef].[PubMed]
    [Google Scholar]
  17. Poxton I. R., McCoubrey J., Blair G.. ( 2001;). The pathogenicity of Clostridium difficile. . Clin Microbiol Infect 7:, 421–427. [CrossRef].[PubMed]
    [Google Scholar]
  18. Tasteyre A., Barc M. C., Collignon A., Boureau H., Karjalainen T.. ( 2001;). Role of FliC and FliD flagellar proteins of Clostridium difficile in adherence and gut colonization. . Infect Immun 69:, 7937–7940. [CrossRef].[PubMed]
    [Google Scholar]
  19. Terao Y., Kawabata S., Kunitomo E., Murakami J., Nakagawa I., Hamada S.. ( 2001;). Fba, a novel fibronectin-binding protein from Streptococcus pyogenes, promotes bacterial entry into epithelial cells, and the fba gene is positively transcribed under the Mga regulator. . Mol Microbiol 42:, 75–86. [CrossRef].[PubMed]
    [Google Scholar]
  20. Voth D. E., Ballard J. D.. ( 2005;). Clostridium difficile toxins: mechanism of action and role in disease. . Clin Microbiol Rev 18:, 247–263. [CrossRef].[PubMed]
    [Google Scholar]
  21. Waligora A. J., Hennequin C., Mullany P., Bourlioux P., Collignon A., Karjalainen T.. ( 2001;). Characterization of a cell surface protein of Clostridium difficile with adhesive properties. . Infect Immun 69:, 2144–2153. [CrossRef].[PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.029553-0
Loading
/content/journal/jmm/10.1099/jmm.0.029553-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error