TY - JOUR
T1 - Enteropathogenic E. coli disrupts tight junction barrier function and structure in vivo
AU - Shifflett, Donnie E.
AU - Clayburgh, Daniel R.
AU - Koutsouris, Athanasia
AU - Turner, Jerrold R.
AU - Hecht, Gail A.
N1 - Funding Information:
This work was presented in preliminary form at the annual meeting of the American Gastroenterological Association in May 2005. We thank Dr Suzana Savkovic for her technical guidance during initiation of this project. This work was supported by grants from the National Institutes of Health #DK50694 and #DK58964 to GH, DK61931 and DK68271 to JRT, and the Department of Veterans Affairs (Merit award) to GH. DRC is a predoctoral fellow of the National Institutes of Health (The University of Chicago Medical Scientist Training Program T32 GM07281).
PY - 2005/10
Y1 - 2005/10
N2 - Enteropathogenic Escherichia coli (EPEC) infection disrupts tight junctions (TJs) and perturbs intestinal barrier function in vitro. E. coli secreted protein F (EspF) is, in large part, responsible for these physiological and morphological alterations. We recently reported that the C57BL/6J mouse is a valid in vivo model of EPEC infection as EPEC colonizes the intestinal epithelium and effaces microvilli. Our current aim was to examine the effects of EPEC on TJ structure and barrier function of the mouse intestine and to determine the role of EspF in vivo. C57BL/6J mice were gavaged with ∼2 × 108 EPEC organisms or PBS. At 1 or 5 days postinfection, mice were killed and ileal and colonic tissue was mounted in Üssing chambers to determine barrier function (measured as transepithelial resistance) and short circuit current. TJ structure was analyzed by immunofluorescence microscopy. Wild-type (WT) EPEC significantly diminished the barrier function of ileal and colonic mucosa at 1 and 5 days postinfection. Deficits in barrier function correlated with redistribution of occludin in both tissues. Infection with an EPEC strain deficient of EspF (ΔespF) had no effect on barrier function at 1 day postinfection. Furthermore, ΔespF had no effect on ileal TJ morphology and minor alterations of colonic TJ morphology at 1 day postinfection. In contrast, at 5 days postinfection, WT EPEC and ΔespF had similar effects on barrier function and occludin localization. In both cases this was associated with immune activation, as demonstrated by increased mucosal tumor necrosis factor-α levels 5 days postinfection. In conclusion, these data demonstrate that WT EPEC infection of 6-8-week-old C57BL/6J mice (1) significantly decreases barrier function in the ileum and colon (2) redistributes occludin in the ileum and colon and (3) is dependent upon EspF to induce TJ barrier defects at early, but not late, times postinfection.
AB - Enteropathogenic Escherichia coli (EPEC) infection disrupts tight junctions (TJs) and perturbs intestinal barrier function in vitro. E. coli secreted protein F (EspF) is, in large part, responsible for these physiological and morphological alterations. We recently reported that the C57BL/6J mouse is a valid in vivo model of EPEC infection as EPEC colonizes the intestinal epithelium and effaces microvilli. Our current aim was to examine the effects of EPEC on TJ structure and barrier function of the mouse intestine and to determine the role of EspF in vivo. C57BL/6J mice were gavaged with ∼2 × 108 EPEC organisms or PBS. At 1 or 5 days postinfection, mice were killed and ileal and colonic tissue was mounted in Üssing chambers to determine barrier function (measured as transepithelial resistance) and short circuit current. TJ structure was analyzed by immunofluorescence microscopy. Wild-type (WT) EPEC significantly diminished the barrier function of ileal and colonic mucosa at 1 and 5 days postinfection. Deficits in barrier function correlated with redistribution of occludin in both tissues. Infection with an EPEC strain deficient of EspF (ΔespF) had no effect on barrier function at 1 day postinfection. Furthermore, ΔespF had no effect on ileal TJ morphology and minor alterations of colonic TJ morphology at 1 day postinfection. In contrast, at 5 days postinfection, WT EPEC and ΔespF had similar effects on barrier function and occludin localization. In both cases this was associated with immune activation, as demonstrated by increased mucosal tumor necrosis factor-α levels 5 days postinfection. In conclusion, these data demonstrate that WT EPEC infection of 6-8-week-old C57BL/6J mice (1) significantly decreases barrier function in the ileum and colon (2) redistributes occludin in the ileum and colon and (3) is dependent upon EspF to induce TJ barrier defects at early, but not late, times postinfection.
KW - EPEC
KW - Enteric pathogen
KW - EspF
KW - Occludin
KW - TNF-α
KW - ZO-1
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U2 - 10.1038/labinvest.3700330
DO - 10.1038/labinvest.3700330
M3 - Article
C2 - 16127426
AN - SCOPUS:25144442798
SN - 0023-6837
VL - 85
SP - 1308
EP - 1324
JO - Laboratory Investigation
JF - Laboratory Investigation
IS - 10
ER -