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.
Highlights
The incubation of IPEC-J2 cells in the porcine colostrum vs. control medium led to a slight improvement in the cell integrity during the first five hours of measurement
We show that the colon epithelium of healthy suckling and weaned piglets and IPEC-J2 cells treated with porcine colostrum are at least for a certain time period protected from C. difficile toxin-induced effects
We have recently demonstrated that neonatal piglets fed a formula milk develop gut symptoms similar to that observed in C. difficile infection (CDI) piglets and they show lower expression of tight junction (TJ) proteins, as compared to suckling piglets [8]
Summary
Toxins A (TcdA) and B (TcdB) besides a binary toxin are the main infection agents leading to loss of epithelial integrity, immune response and intestinal damage [3] Their action is related to the modulation of the intestinal epithelial cell physiology and disruption of barrier function via inactivation of Rho proteins involved in the formation of the cytoskeleton. This leads to disruption of tight junction (TJ) proteins and to a loss of epithelial integrity, as demonstrated in porcine and human cell lines [4,5,6,7]. The reasons why some piglets of the same litter get sick and others do not remain unknown so far
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