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Effects of Ex Vivo Infection with ETEC on Jejunal Barrier Properties and Cytokine Expression in Probiotic-Supplemented Pigs

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Abstract

Background and Aim

Enterotoxigenic Escherichia coli (ETEC) strains are involved in piglet post-weaning diarrhea. Prophylactic measures including probiotics have been examined in infection experiments with live piglets. In the present study, we have tested whether the early effects of ETEC infection can also be evoked and studied in a model in which ETEC is added to whole mucosal tissues ex vivo, and whether this response can be modulated by prior supplementation of the piglets with probiotics.

Methods

Jejunal barrier and transport properties of Enterococcus faecium-supplemented or control piglets were assessed in Ussing chambers. Part of the epithelia was challenged with an ETEC strain at the mucosal side. Fluxes of fluorescein as a marker of paracellular permeability, and the expression of selected tight junction (TJ) proteins and of proinflammatory cytokines were measured.

Results

The addition of ETEC ex vivo induced an increase in transepithelial resistance peaking in the first 2 h with a concomitant reduction in fluorescein fluxes, indicating tightening effects on barrier function. The response of short-circuit current after stimulation with PGE2 or glucose was reduced in epithelia treated with ETEC. ETEC induced a decrease in the TJ protein claudin-4 in the control diet group after 280 min and an increase in the mRNA expression of the proinflammatory cytokines interleukin-8 and TNF-α in both groups after 180 min.

Conclusions

The addition of ETEC ex vivo affected barrier function and transport properties of the jejunal tissues and enhanced cytokine expression. The differences in claudin-4 expression in the jejunum might indicate a beneficial effect of E. faecium prefeeding.

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Acknowledgments

We thank K. Wolf, U. Tietjen, and G. Greco for technical support, and K. Wolf for help with the analysis of data. This research was financially supported by the German Research Foundation, Grant No. SFB 852/2 and LO 2058/1-1.

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Authors and Affiliations

  1. Institute of Veterinary Physiology, Department of Veterinary Medicine, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany

    Ulrike Lodemann, Salah Amasheh, Judith Radloff, Martina Kern & Jörg R. Aschenbach

  2. Institute of Microbiology and Epizootics, Department of Veterinary Medicine, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany

    Astrid Bethe & Lothar H. Wieler

  3. Robert Koch Institute, Nordufer 20, 13302, Berlin, Germany

    Lothar H. Wieler

  4. Institute of Animal Nutrition, Department of Veterinary Medicine, Freie Universität Berlin, Königin-Luise-Str. 49, 14195, Berlin, Germany

    Robert Pieper & Jürgen Zentek

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  1. Ulrike Lodemann
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  2. Salah Amasheh
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Correspondence to Ulrike Lodemann.

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Lodemann, U., Amasheh, S., Radloff, J. et al. Effects of Ex Vivo Infection with ETEC on Jejunal Barrier Properties and Cytokine Expression in Probiotic-Supplemented Pigs. Dig Dis Sci 62, 922–933 (2017). https://doi.org/10.1007/s10620-016-4413-x

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