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Difference in the biological effects of Clostridium difficile toxin B in proliferating and non-proliferating cells

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Abstract

Toxin A (TcdA) and toxin B (TcdB) from Clostridium difficile are the causative agents of the C. difficile-associated diarrhea (CDAD) and its severe form, the pseudomembranous colitis. TcdA and TcdB both glucosylate and thereby inactivate low molecular weight GTP-binding proteins of the Rho, Rac, and Cdc42 subfamilies. In cultured cell lines, TcdB induces actin re-organization and bi-nucleation (“cytopathic effects”) and cell death (“cytotoxic effects”). In this study, the role of cell cycle progression in the cytopathic and the cytotoxic effects of TcdB is evaluated by a differential analysis of these effects in proliferating and non-proliferating cells. Density-synchronized murine fibroblasts and confluent HT29 colonocytes are exploited as cell culture models for non-proliferating cells. Cell death is analyzed in terms of a loss of cell viability, phosphatidylserine exposure, and DNA fragmentation. In proliferating cells, TcdB blocks cell proliferation and induces apoptotic cell death. In contrast, TcdB induces non-apoptotic cell death in non-proliferating cells. TcdB-induced cell rounding turns out to be independent of cell cycle progression. Cell cycle progression is an important determinant in the biological effects of TcdB. With respect to the pathology of CDAD, this study leads to the new hypothesis that necrotic cell death of terminally differentiated colonocytes and inhibition of epithelial renewal of the colon contribute to the pathogenesis of CDAD.

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Abbreviations

CDAD:

C. difficile-associated diarrhea

PMC:

Pseudomembranous colitis

TcdA:

Toxin A from the C. difficile strain VPI10463

TcdB:

Toxin B from the C. difficile strain VPI10463

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (priority program 1150 grant GE 1247/1-3).

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Authors and Affiliations

  1. Institut für Toxikologie, Medizinische Hochschule Hannover, 30625, Hannover, Germany

    Marta Lica, Florian Schulz, Ilona Schelle, Martin May, Ingo Just & Harald Genth

  2. Institut für Toxikologie OE 5340, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Harald Genth

Authors
  1. Marta Lica
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  2. Florian Schulz
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  3. Ilona Schelle
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  4. Martin May
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  5. Ingo Just
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  6. Harald Genth
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Correspondence to Harald Genth.

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This article is published as part of the special issue on bacterial toxins.

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Lica, M., Schulz, F., Schelle, I. et al. Difference in the biological effects of Clostridium difficile toxin B in proliferating and non-proliferating cells. Naunyn-Schmied Arch Pharmacol 383, 275–283 (2011). https://doi.org/10.1007/s00210-010-0595-5

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  • DOI: https://doi.org/10.1007/s00210-010-0595-5

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