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Epithelial barrier dysfunction in lymphocytic colitis through cytokine-dependent internalization of claudin-5 and -8

  • Original Article—Alimentary Tract
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Abstract

Background

Watery diarrhea is the cardinal symptom of lymphocytic colitis (LC). We have previously shown that colonic Na malabsorption is one of the major pathologic alterations of LC and found evidence for an epithelial barrier defect. On these grounds, this study aimed to identify the inherent mechanisms of this epithelial barrier dysfunction and its regulatory features.

Methods

Epithelial resistance (R epi) was determined by one-path impedance spectroscopy and 3H-mannitol fluxes were performed on biopsies from sigmoid colon in miniaturized Ussing chambers. Tight junction proteins were analyzed by Western blot and confocal microscopy. Inflammatory signaling was characterized in HT-29/B6 cells. Apoptosis and mucosal surface parameters were quantified morphologically.

Results

R epi was reduced to 53% and 3H-mannitol fluxes increased 1.7-fold in LC due to lower expression of claudin-4, -5, and -8 and altered subcellular claudin-5 and -8 distributions off the tight junction. TNFα and IFNγ could mimic subcellular redistribution in HT-29/B6 cells, a process which was independent on MLCK activation. Epithelial apoptosis did not contribute to barrier dysfunction in LC and mucosal surface area was unchanged.

Conclusions

Epithelial barrier dysfunction in LC occurs through downregulation of claudin-4, -5, and -8, and redistribution of claudin-5 and -8 off the tight junction, which contributes to diarrhea by a leak-flux mechanism. The key effector cytokines TNFα and IFNγ turned out to be the trigger for redistribution of claudin-5 and -8. Thus, alongside sodium malabsorption, leak-flux is yet another important diarrheal mechanism in LC.

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Acknowledgments

We thank Detlef Sorgenfrei († January 2015) for excellent technical support.

Author information

Author notes

  1. Christoph Loddenkemper

    Present address: Institute of Pathology PathoTres, Berlin, Germany

Authors and Affiliations

  1. Department of Gastroenterology, Infectious Diseases and Rheumatology, Charité, Campus Benjamin Franklin, Berlin, Germany

    Christian Barmeyer, Irene Erko, Christian Bojarski, Svenja Meissner, Britta Siegmund & Jörg-Dieter Schulzke

  2. Institute of Clinical Physiology, Charité, Campus Benjamin Franklin, Hindenburgdamm 30, 12203, Berlin, Germany

    Christian Barmeyer, Karem Awad, Anja Fromm, Michael Fromm & Jörg-Dieter Schulzke

  3. Institute of Pathology, Charité, Campus Benjamin Franklin, Berlin, Germany

    Christoph Loddenkemper

  4. Max Planck Institute for Molecular Genetics, Berlin, Germany

    Martin Kerick & Michal R. Schweiger

  5. Cologne Center for Genomics, University of Cologne, Cologne, Germany

    Michal R. Schweiger

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  1. Christian Barmeyer
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Correspondence to Jörg-Dieter Schulzke.

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Funding information

This work was supported by the Deutsche Forschungsgemeinschaft (DFG) Grants Schu 559/10-2, FOR 721/2 and the Volkswagenstiftung (Lichtenberg Program to MRS).

Conflict of interest

Britta Siegmund has served as consultant for Abbvie, Janssen, Hospira, MSD, Mundipharma, Takeda, received speaker fees from Abbvie, Falk, Ferring, Hospira, MSD, Takeda and received a research grant from Hospira. All other authors declare that they have no conflicts of interest.

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Barmeyer, C., Erko, I., Awad, K. et al. Epithelial barrier dysfunction in lymphocytic colitis through cytokine-dependent internalization of claudin-5 and -8. J Gastroenterol 52, 1090–1100 (2017). https://doi.org/10.1007/s00535-017-1309-2

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  • DOI: https://doi.org/10.1007/s00535-017-1309-2

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