Yersinia enterocolitica induces epithelial barrier dysfunction through regional tight junction changes in colonic HT-29/B6 cell monolayers.
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 Ω cm² (P<0.001) 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.
Department of Gastroenterology, Infectious Diseases and Rheumatology, Charité Campus Benjamin Franklin, Berlin, Germany.
Nina A
NA
Hering
Jan F
JF
Richter
Susanne M
SM
Krug
Dorothee
D
Günzel
Anja
A
Fromm
Erwin
E
Bohn
Rita
R
Rosenthal
Roland
R
Bücker
Michael
M
Fromm
Hanno
H
Troeger
Jörg D
JD
Schulzke
D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D057167:Claudins / Q000378:metabolism; D003106:Colon / Q000166:cytology* / Q000382:microbiology; D003967:Diarrhea / Q000382:microbiology*; D058266:Dielectric Spectroscopy; D017097:Electric Impedance; D004848:Epithelium / Q000382:microbiology / Q000473:pathology*; D019793:Fluorescein / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D020287:In Situ Nick-End Labeling; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008353:Mannitol / Q000378:metabolism; D008854:Microscopy, Electron; D009336:Necrosis / Q000473:pathology*; D010539:Permeability; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019108:Tight Junctions / Q000473:pathology*; D015008:Yersinia enterocolitica / Q000502:physiology*
D057167:Claudins; D008353:Mannitol; D007770:L-Lactate Dehydrogenase; D002118:Calcium; D019793:Fluorescein