TY - JOUR
T1 - Notch-1 signaling regulates intestinal epithelial barrier function, through interaction with CD4+ T cells, in mice and humans
AU - Dahan, Stephanie
AU - Rabinowitz, Keren M.
AU - Martin, Andrea P.
AU - Berin, M. Cecilia
AU - Unkeless, Jay C.
AU - Mayer, Lloyd
N1 - Funding Information:
Funding Supported by the Crohn's and Colitis Foundation of America (2442) and National Institutes of Health grants AI044236 , AI084952 , DK072201 , and DK086605 . Confocal laser scanning microscopy was performed at the MSSM-Microscopy Shared Resource Facility, supported by funding from National Institutes of Health/National Cancer Institute–shared resources grant 5R24 CA095823-04 , National Science Foundation Major Research Instrumentation grant DBI-9724504 , and National Institutes of Health–shared instrumentation grant 1 S10 RR0 9145-01 .
PY - 2011/2
Y1 - 2011/2
N2 - Background & Aims Interactions between lymphocytes and intestinal epithelial cells occur in the subepithelial space of the gastrointestinal tract. Normal human lamina propria lymphocytes (LPLs) induce differentiation of intestinal epithelial cells. The absence of LPLs in mice, such as in RAG1 -/- mice, results in defects in epithelial cell differentiation. We investigated the role of lymphoepithelial interactions in epithelial differentiation and barrier function. Methods We used adoptive transfer to determine if CD4+ T cells (CD4+CD62L +CD45RbHi and/or CD4+CD62L +CD45RbLo) could overcome permeability defect (quantified in Ussing chambers). Immunofluorescence staining was performed to determine expression of cleaved Notch-1, villin, and claudin 5 in colon samples from mice and humans. Caco-2 cells were infected with a lentivirus containing a specific Notch-1 or scrambled short hairpin RNA sequence. Tight junction assembly was analyzed by immunoblot and immunofluorescence analyses, and transepithelial resistance was monitored. Results Expression of cleaved Notch-1, villin, or claudin 5 was not detected in RAG1-/- colonocytes; their loss correlated with increased intestinal permeability. Transfer of CD45Rb Hi and/or CD45RbLo cells into RAG1-/- mice induced expression of cleaved Notch, villin, and claudin 5 in colonocytes and significantly reduced the permeability of the distal colon. Loss of Notch-1 expression in Caco-2 cells correlated with decreased transepithelial resistance and dysregulated expression and localization of tight junction proteins. Levels of cleaved Notch-1 were increased in colonic epithelium of patients with Crohn's disease. Conclusions LPLs promote mucosal barrier function, which is associated with activation of the Notch-1 signaling pathway. LPLs maintain intestinal homeostasis by inducing intestinal epithelial cell differentiation, polarization, and barrier function.
AB - Background & Aims Interactions between lymphocytes and intestinal epithelial cells occur in the subepithelial space of the gastrointestinal tract. Normal human lamina propria lymphocytes (LPLs) induce differentiation of intestinal epithelial cells. The absence of LPLs in mice, such as in RAG1 -/- mice, results in defects in epithelial cell differentiation. We investigated the role of lymphoepithelial interactions in epithelial differentiation and barrier function. Methods We used adoptive transfer to determine if CD4+ T cells (CD4+CD62L +CD45RbHi and/or CD4+CD62L +CD45RbLo) could overcome permeability defect (quantified in Ussing chambers). Immunofluorescence staining was performed to determine expression of cleaved Notch-1, villin, and claudin 5 in colon samples from mice and humans. Caco-2 cells were infected with a lentivirus containing a specific Notch-1 or scrambled short hairpin RNA sequence. Tight junction assembly was analyzed by immunoblot and immunofluorescence analyses, and transepithelial resistance was monitored. Results Expression of cleaved Notch-1, villin, or claudin 5 was not detected in RAG1-/- colonocytes; their loss correlated with increased intestinal permeability. Transfer of CD45Rb Hi and/or CD45RbLo cells into RAG1-/- mice induced expression of cleaved Notch, villin, and claudin 5 in colonocytes and significantly reduced the permeability of the distal colon. Loss of Notch-1 expression in Caco-2 cells correlated with decreased transepithelial resistance and dysregulated expression and localization of tight junction proteins. Levels of cleaved Notch-1 were increased in colonic epithelium of patients with Crohn's disease. Conclusions LPLs promote mucosal barrier function, which is associated with activation of the Notch-1 signaling pathway. LPLs maintain intestinal homeostasis by inducing intestinal epithelial cell differentiation, polarization, and barrier function.
KW - Immune System
KW - Inflammatory Bowel Disease
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UR - http://www.scopus.com/inward/citedby.url?scp=79251537479&partnerID=8YFLogxK
U2 - 10.1053/j.gastro.2010.10.057
DO - 10.1053/j.gastro.2010.10.057
M3 - Article
C2 - 21056041
AN - SCOPUS:79251537479
SN - 0016-5085
VL - 140
SP - 550
EP - 559
JO - Gastroenterology
JF - Gastroenterology
IS - 2
ER -
