Disintegration of the colonic epithelial barrier is considered a key event in the initiation and progression of inflammatory bowel and celiac disease. As the primary etiology of these diseases remains unknown, we hypothesized that the trichothecene deoxynivalenol (DON), a fungal metabolite found in grain-based human diets, might be one of the triggers resulting in an impairment of the intestinal tight junction network preceding an inflammatory response. Using horizontal impedance measurements, we demonstrate that DON disintegrates a human Caco-2 cell monolayer within <1 h after exposure to concentrations as low as 1.39 μM. This initial trigger is followed by a decrease in transepithelial resistance and an increased permeability of marker molecules, such as lucifer yellow and FITC-labeled dextran. In parallel, the increase in paracellular transport of FITC-dextran is demonstrated in vivo in B6C3F1 mice, challenged orally with DON. In vitro claudin protein levels are decreased and correlated with a displacement within the cells in vitro and in vivo, accompanied by a compensatory up-regulation of mRNA levels of claudins and their binding partner ZO-1. In treated mice, alterations in villus architecture in the entire intestinal tract resemble the disintegration of the epithelial barrier, a characteristic of chronic inflammatory bowel disease.-Akbari, P., Braber, S., Gremmels, H., Koelink, P. J., Verheijden, K. A. T., Garssen, J., Fink-Gremmels, J. Deoxynivalenol: a trigger for intestinal integrity breakdown.