Molecular mechanisms underlying the probiotic effects of Escherichia coli Nissle 1917 involve ZO-2 and PKCζ redistribution resulting in tight junction and epithelial barrier repair

Authors

  • Agata A. Zyrek,

    1. Institut für Infektiologie, Zentrum für Molekularbiologie der Entzündung (ZMBE), Westfälische Wilhelms-Universität/Universitätsklinikum Münster, von-Esmarch-Str. 56, D-48149 Münster, Germany.
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  • Christoph Cichon,

    1. Institut für Infektiologie, Zentrum für Molekularbiologie der Entzündung (ZMBE), Westfälische Wilhelms-Universität/Universitätsklinikum Münster, von-Esmarch-Str. 56, D-48149 Münster, Germany.
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  • Simone Helms,

    1. Institut für Infektiologie, Zentrum für Molekularbiologie der Entzündung (ZMBE), Westfälische Wilhelms-Universität/Universitätsklinikum Münster, von-Esmarch-Str. 56, D-48149 Münster, Germany.
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  • Corinne Enders,

    1. Ardeypharm GmbH, Abteilung für Biologische Forschung, Loerfeldstr. 20, D-59313 Herdecke, Germany.
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  • Ulrich Sonnenborn,

    1. Ardeypharm GmbH, Abteilung für Biologische Forschung, Loerfeldstr. 20, D-59313 Herdecke, Germany.
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  • M. Alexander Schmidt

    Corresponding author
    1. Institut für Infektiologie, Zentrum für Molekularbiologie der Entzündung (ZMBE), Westfälische Wilhelms-Universität/Universitätsklinikum Münster, von-Esmarch-Str. 56, D-48149 Münster, Germany.
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*E-mail infekt@uni-muenster.de; Tel. (+49) 251 8356466; Fax (+49) 251 8356467.

Summary

The probiotic Escherichia coli strain Nissle 1917 (EcN) has been used for decades in human medicine in Central Europe for the treatment and prevention of intestinal disorders and diseases. However, the molecular mechanisms underlying its beneficial effects are only partially understood. To identify molecular responses induced by EcN that might contribute to its probiotic properties polarized T84 cells were investigated employing DNA microarrays, quantitative RT-PCR, Western blotting, immunofluorescence and specific protein kinase C (PKC) inhibitors. Polarized T84 epithelial cell monolayers were used as a model to monitor barrier disruption by infection with the enteropathogenic E. coli (EPEC) strain E2348/69. Co-incubation of EPEC with EcN or addition of EcN following EPEC infection abolished barrier disruption and, moreover, restored barrier integrity as monitored by transepithelial resistance. DNA-microarray analysis of T84 cells incubated with EcN identified 300+ genes exhibiting altered expression. EcN altered the expression, distribution of zonula occludens-2 (ZO-2) protein and of distinct PKC isotypes. ZO-2 expression was enhanced in parallel to its redistribution towards the cell boundaries. This study provides evidence that EcN induces an overriding signalling effect leading to restoration of a disrupted epithelial barrier. This is transmitted via silencing of PKCζ and the redistribution of ZO-2. We suggest that these properties contribute to the reported efficacy in the treatment of inflammatory bowel diseases and in part rationalize the probiotic nature of EcN.

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