Epithelial myosin light chain kinase-dependent barrier dysfunction mediates T cell activation-induced diarrhea in vivo.
Disruption of the intestinal epithelial barrier occurs in many intestinal diseases, but neither the mechanisms nor the contribution of barrier dysfunction to disease pathogenesis have been defined. We utilized a murine model of T cell-mediated acute diarrhea to investigate the role of the epithelial barrier in diarrheal disease. We show that epithelial barrier dysfunction is required for the development of diarrhea. This diarrhea is characterized by reversal of net water flux, from absorption to secretion; increased leak of serum protein into the intestinal lumen; and altered tight junction structure. Phosphorylation of epithelial myosin II regulatory light chain (MLC), which has been correlated with tight junction regulation in vitro, increased abruptly after T cell activation and coincided with the development of diarrhea. Genetic knockout of long myosin light chain kinase (MLCK) or treatment of wild-type mice with a highly specific peptide MLCK inhibitor prevented epithelial MLC phosphorylation, tight junction disruption, protein leak, and diarrhea following T cell activation. These data show that epithelial MLCK is essential for intestinal barrier dysfunction and that this barrier dysfunction is critical to pathogenesis of diarrheal disease. The data also indicate that inhibition of epithelial MLCK may be an effective non-immunosuppressive therapy for treatment of immune-mediated intestinal disease.
Clayburgh
Daniel R
DR
Department of Pathology, The University of Chicago, Chicago, Illinois, USA.
Barrett
Terrence A
TA
Tang
Yueming
Y
Meddings
Jon B
JB
Van Eldik
Linda J
LJ
Watterson
D Martin
DM
Clarke
Lane L
LL
Mrsny
Randall J
RJ
Turner
Jerrold R
JR
D000818:Animals; D001327:Autoimmune Diseases / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D001798:Blood Proteins / Q000378:metabolism; D003967:Diarrhea / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D007413:Intestinal Mucosa / Q000378:metabolism* / Q000648:ultrastructure; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D018345:Mice, Knockout; D018994:Myosin Light Chains / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000235:genetics / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism* / Q000648:ultrastructure; D019108:Tight Junctions / Q000378:metabolism / Q000648:ultrastructure; D014867:Water / Q000378:metabolism
D001798:Blood Proteins; D018994:Myosin Light Chains; D014867:Water; D009219:Myosin-Light-Chain Kinase