Epithelial NF- (cid:2) B Enhances Transmucosal Fluid Movement by Altering Tight Junction Protein Composition after T Cell Activation

In inflammatory bowel disease (IBD), aberrant activation of innate and adaptive immune responses enhances mucosal permeability through mechanisms not completely understood. To examine the role of epithelial nuclear factor (NF- (cid:2) B) in IBD-induced enhanced permeability, epithelial-specific I (cid:2) B (cid:3) mutant (NF- (cid:2) B super repressor) transgenic (TG) mice were generated. NF-kB activation was inhibited in TG mice, relative to wild-type mice, following T cell-mediated immune cell activation using an anti-CD3 monoclonal antibody. Furthermore, epithelial NF- (cid:2) B super repressor protein inhibited diarrhea and blocked changes in transepithelial resistance and transmucosal flux of alexa350 (0.35 kDa) and dextran3000 (3 kDa). In vivo perfusion loop studies in TG mice revealed reversed net water secretion and reduced lumenal flux of different molecular probes (bovine serum albumin, alexa350, and dextran3000). T cell activation was induced via i.p. injection of a T cell receptor cross-linking monoclonal antibody (mAb) specific for the CD3 (cid:5) subunit. Anti-CD3 has been used in multiple in vivo studies to examine T cell activation on epithelial apoptosis, intestinal permeability, and fluid loss. Data from anti-CD3-treated wild-type and TNF recep-tor-1 knockout mice indicate that T cell-induced TNF reduces sodium and water absorption by epithelial cells, in part, by inhibiting epithelial Na/K ATPase. Further reveal that epithelial myosin light chain kinase (MLCK) activation contributes to T cell-induced diarrhea by reducing Analyses of human tissue confirms that epithelial Na/K ATPase activity, reduced and water and elevated myosin light chain phosphorylation, are consistent features of IBD compared with normal tissue. 3,14 Data from the current study suggest that T cell-induced NF- (cid:2) B activation opens paracellular spaces and enhances transmucosal fluid movement by altering TJ protein composition. Our findings support view that resulting from NF- (cid:2) B-mediated changes of epithelial TJs important for mucosal host defense

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