Enteropathogenic Escherichia coli adherence to intestinal epithelial monolayers diminishes barrier function.
The mechanism by which enteropathogenic Escherichia coli (EPEC) causes diarrhea remains elusive. Several alterations within the host cell have been demonstrated to occur following EPEC attachment including increases in intracellular Ca2+ concentration and rearrangement and phosphorylation of several cytoskeletal proteins. The consequences of these intracellular perturbations on host cell function, however, have not been determined. The aim of this study was to examine the effect of EPEC adherence on intestinal epithelial barrier function. T84 cell monolayers were infected with either wild-type EPEC or a nonadherent isogenic derivative. Transepithelial electrical resistance, a measure of barrier function, decreased 33.5 +/- 6.4% after a 6-h incubation with the wild-type strain. Electron microscopy revealed ultrastructurally normal cells, and lactate dehydrogenase release assays failed to demonstrate cytotoxicity. Dual 22Na+ and [3H]mannitol flux studies localized the permeability defect to tight junctions. In addition, cumulative flux of the paracellular marker mannitol was four- to fivefold greater across monolayers infected with wild-type EPEC. Sequestration of intracellular calcium stores by dantrolene completely abrogated the resistance drop associated with EPEC attachment. These data demonstrate that adherence of EPEC to intestinal epithelial cell monolayers disrupts tight junction barrier function via a calcium-requiring event.
Spitz
J
J
Department of Medicine (Gastroenterology, University of Illinois at Chicago.
Yuhan
R
R
Koutsouris
A
A
Blatt
C
C
Alverdy
J
J
Hecht
G
G
D001422:Bacterial Adhesion; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D004553:Electric Conductivity; D004926:Escherichia coli / Q000502:physiology*; D007413:Intestinal Mucosa / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D007425:Intracellular Membranes / Q000378:metabolism; D008854:Microscopy, Electron; D010539:Permeability
D002118:Calcium