The Role of Flagella in Clostridium difficile Pathogenesis: Comparison between a Non-Epidemic and an Epidemic Strain
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1 The Role of Flagella in Clostridium difficile Pathogenesis: Comparison between a Non-Epidemic and an Epidemic Strain Soza T. Baban, Sarah A. Kuehne, Amira Barketi-Klai, Stephen T. Cartman, Michelle Kelly, Kim R. Hardie, Imad Kansau, Anne Collignon, Nigel P. Minton Clostridia Research Group NIHR Biomedical Research Unit in GI Disease Centre for Biomolecular Sciences The University of Nottingham United Kingdom
2 Background- Clostridium difficile- the organism and the disease Clostridium difficile- The worldwide leading cause of hospital-associated infections. The incidence and severity of CDI continue to increase particularly following the global emergence of epidemic hypervirulent C. difficile strains (027/BI/NAP1). The prevention and control of C. difficile infection in health care settings has become a global public health challenge.
3 C. difficile virulence factors C. difficile produce two major toxins, toxin A and toxin B, both of which are responsible of the clinical manifestations of the disease. C. difficile expresses multiple surface adhesins, potentially functioning as colonization factors.
4 Role of flagella & motility in virulence Flagella function as an adhesin in mediating adherence of C. difficile bacteria to intestinal epithelial cells. Potential functions for flagella in virulence: Motility apparatus Surface adhesins and invasins Secretion apparatus
5 Aims of this study To elucidate the mechanism by which flagella contribute to C. difficile adhesion to human intestinal epithelial cells To assess the contribution of flagellum-mediated motility in colonisation of the intestinal tract in mice Does some part of the polar flagellum act as an adhesin, or is it active motility that is needed for adherence and intestinal colonization?
6 Construction and phenotypic characterization of flagella-associated mutants Southern blot analysis
7 Phenotypic characterization of flagella-associated mutants for motility
8 Phenotypic characterization of flagella-associated mutants for flagellation
9 Importance of flagella of R20291 for in vitro cell adherence The flagellum of R20291 is used as an adhesin to enhance bacterial association to human intestinal epithelial cells in vitro
10 Importance of flagella of C. difficile 630Δerm for in vitro cell adherence Surprisingly, all flagellar mutants adhered significantly more strongly to human intestinal caco-2 cells than the wild-type strains.
11 In vivo analysis: Importance of flagellum of R20291 strain in intestinal colonization of axenic mice Monoxenic mouse model of colonization (Individual mouse infections)
12 Dixenic mouse model of colonization (coinfection study)
13 Dixenic mouse model of colonization (coinfection study)
14 In vivo analysis: Importance of flagellum of C. difficile 630Δerm strain in intestinal colonization of axenic mice Human Microbial Flora mouse model
15 Monoxenic mouse model of colonization (Individual mouse infections)
16 Dixenic mouse model of colonization (coinfection study)
17 Conclusion The flagellum of R20291 functions as a surface adhesin, not as motility apparatus to enhance bacterial association to human intestinal epithelial cells and establish intestinal colonization. Flagella of C. difficile 630Δerm are not required for adherence and intestinal colonisation in this strain, but flagella-mediated motility might contribute to an overall fitness of the bacteria. The flagellum plays an important role as a virulence factor in contributing to the hypervirulence of the epidemic C. difficile R20291 which causes more severe infections than historical strain.
18 Acknowledgments Prof. Nigel P. Minton Dr. Kim R. Hardie Dr. Sarah A. Kuehne Dr. Stephen T. Cartman Michelle Kelly Prof. Anne Collignon Dr. Imad Kansau Dr. Amira Barketi-Klai
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