Yersinia enterocolitica induces epithelial barrier dysfunction through regional tight junction changes in colonic HT-29/B6 cell monolayers

Abstract

Yersinia enterocolitica is a common cause of acute gastroenteritis. This study aimed to clarify the mechanisms leading to barrier dysfunction and diarrhea. Exposure of human colonic HT-29/B6 cells to Y. enterocolitica resulted in a decrease in transepithelial resistance from 404 ± 23 to 163 ± 21 O cm 2 (Po0.001) in parallel with an increase in mannitol (182 Da) and fluorescein (332 Da) permeability, whereas short circuit current did not change. This effect was time dependent, required the presence of living bacteria, could not be triggered by bacterial supernatants and was not due to Yersinia outer proteins. Concomitantly, Y. enterocolitica induced necrosis as indicated by an increase in lactate dehydrogenase-release, whereas epithelial apoptosis was not upregulated. Local changes in conductivity were detected by conductance scanning, indicating 'leaky regions' within the epithelium that were visualized by biotinylation and confocal microscopy. In these regions, claudin-3 and -4 and, especially claudin-8, were redistributed off the tight junction (TJ) into the cytoplasm. In addition, the expression of claudin-2, -3, -8, -10 and ZO-1 was diminished as quantified by immunoblotting. Moreover, we found claudin-8 to be regulated by the c-Jun N-terminal kinase, the inhibition of which attenuated the Y. enterocolitica-induced decrease in transepithelial resistance and restored claudin-8 protein level. In conclusion, barrier dysfunction in Y. enterocolitica infection is due to circumscribed epithelial TJ protein changes and necrotic cell loss, as a consequence of which leak flux diarrhea and antigen-uptake provoking extraintestinal arthritis may be triggered.

Claudin-2 as a mediator of leaky gut barrier during intestinal inflammation

“…One possibility is that impaired production of antimicrobial defensins associated with CARD15/NOD2 mutations could facilitate bacterial translocation into the mucosa, leading to inflammation and cytokine-mediated TJ changes. [72][73][74][75][76] In support of this thesis, CARD15/NOD2 knockout mice, which are more susceptible to TNBS colitis, show intestinal changes that include enhanced bacterial translocation, elevated TNFa, IFNg, and IL-4, decreased ZO-1 and -2, and increased permeability. 77 Although potential changes in the expression of claudin proteins were not reported, all 3 cytokines are known to have significant effects on TJ protein expression, including the regulation of claudin-2.…”

Toxoplasma gondii Infection Promotes Epithelial Barrier Dysfunction of Caco-2 Cells

“…38,39 Also, Yersinia enterocolitica induces epithelial barrier dysfunction through regional TJ changes. 40 These findings suggest that TJ proteins are frequent targets of intestinal pathogens in the process of invasion and infection. It is known that NF-κB enhances the permeability of T84 or Caco-2 cells, decreasing the levels of TJ proteins and disturbance in TJ localization.…”

Germ‐free mice as a model to study effect of gut microbiota on host physiology

“…4 Yersinia enterocolitica is associated with altered tight junction proteins and induction of cell necrosis, both resulting in a decrease in transepithelial resistance in colonic HT-29/B6 cell monolayers. 5 Entamoeba histolytica infection causes constitutive production and secretion of prostaglandin E 2 which results in altered paracellular permeability of T84 monolayers resulting in increased sodium permeability and chloride secretion by activation of cystic fibrosis transmembrane conductance regulator. 6 Campylobacter infection disrupts barrier function, with decreased transepithelial electrical resistance and a change in the distribution of the tight junction protein occludin within Caco-2 cell monolayers.…”