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Porcine Colostrum Protects the IPEC-J2 Cells and Piglet Colon Epithelium against Clostridioides (syn. Clostridium ) difficile Toxin-Induced Effects

Authors
  • Grześkowiak, Łukasz1
  • Pieper, Robert1
  • Kröger, Susan1
  • Martínez-Vallespín, Beatriz1
  • Hauser, Anja E.2, 3
  • Niesner, Raluca2, 4
  • Vahjen, Wilfried1
  • Zentek, Jürgen1
  • 1 Institute of Animal Nutrition, Freie Universität Berlin, 14195 Berlin, Germany
  • 2 German Rheumatism Research Center, Berlin, A Leibniz Institute, 10117 Berlin, Germany
  • 3 Intravital Microscopy and Immune Dynamics, Charité–Universitätsmedizin Berlin, 10117 Berlin, Germany
  • 4 Dynamic and Functional in vivo Imaging, Veterinary Medicine, Freie Universität Berlin, 14163 Berlin, Germany
Type
Published Article
Journal
Microorganisms
Publisher
MDPI AG
Publication Date
Jan 20, 2020
Volume
8
Issue
1
Identifiers
DOI: 10.3390/microorganisms8010142
PMID: 31968636
PMCID: PMC7022787
Source
PubMed Central
Keywords
License
Green

Abstract

Clostridioides difficile toxins are one of the main causative agents for the clinical symptoms observed during C. difficile infection in piglets. Porcine milk has been shown to strengthen the epithelial barrier function in the piglet’s intestine and may have the potential to neutralise clostridial toxins. We hypothesised that porcine colostrum exerts protective effects against those toxins in the IPEC-J2 cells and in the colon epithelium of healthy piglets. The IPEC-J2 cells were treated with either the toxins or porcine colostrum or their combination. Analyses included measurement of trans-epithelial electrical resistance (TEER), cell viability using propidium iodide by flow cytometry, gene expression of tight junction (TJ) proteins and immune markers, immunofluorescence (IF) histology of the cytoskeleton and a TJ protein assessment. Colon tissue explants from one- and two-week-old suckling piglets and from five-week-old weaned piglets were treated with C. difficile toxins in Ussing chamber assays to assess the permeability to macromolecules (FITC-dextran, HRP), followed by analysis of gene expression of TJ proteins and immune markers. Toxins decreased viability and integrity of IPEC-J2 cells in a time-dependent manner. Porcine colostrum exerted a protective effect against toxins as indicated by TEER and IF in IPEC-J2 cells. Toxins tended to increase paracellular permeability to macromolecules in colon tissues of two-week-old piglets and downregulated gene expression of occludin in colon tissues of five-week-old piglets ( p = 0.05). Porcine milk including colostrum, besides other maternal factors, may be one of the important determinants of early immune programming towards protection from C. difficile infections in the offspring.

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