title: Interleukin-18 facilitates neutrophil transmigration via myosin light chain kinase-dependent disruption of occludin, without altering epithelial permeability.
abstract: Compromised epithelial barrier function and tight junction alterations are hallmarks of a number of gastrointestinal disorders, including inflammatory bowel disease (IBD). Increased levels of IL-18 have been observed in mucosal samples from Crohn's disease and ulcerative colitis patients. Remarkably, several reports have demonstrated that immunological or genetic blockage of IL-18 ameliorates the severity of colitis in multiple in vivo models of IBD. Nevertheless, the effects of IL-18 on intestinal epithelial barrier function remain unclear. We hypothesized that IL-18 could disrupt intestinal epithelial barrier structure and function, thus contributing to tissue damage in the context of IBD. The aims of the present study were to determine the effects of IL-18 on epithelial barrier structure and function and to characterize the mechanisms involved in these modulatory properties. Human colonic epithelial Caco-2 monolayers were coincubated with IL-18 for 24 h and processed for immunocytochemistry, immunoblotting, quantitative PCR, and permeability measurements (transepithelial resistance, FITC-dextran fluxes, and bacterial translocation). Our findings indicate that IL-18 selectively disrupts tight junctional occludin, without affecting the distribution pattern of claudin-4, claudin-5, zonula occludens-1, or E-cadherin. This effect coincided with a significant increase in myosin light chain kinase (MLCK) protein levels and activity. Pharmacological inhibition of MLCK and NF-κB prevented IL-18-induced loss of occludin. Although too subtle to alter paracellular permeability, these fine changes correlated with an MLCK-dependent increase in neutrophil transepithelial migration. In conclusion, our data suggest that IL-18 may potentiate inflammation in the context of IBD by facilitating neutrophil transepithelial migration via MLCK-dependent disruption of tight junctional occludin.
authors.affiliation: Dept. of Biological Sciences, Univ. of Calgary, Calgary, AB, Canada.
authors.forename: Tamia K
authors.initials: TK
authors.lastname: Lapointe
authors.forename: Andre G
authors.initials: AG
authors.lastname: Buret
mesh_terms: D001381:Azepines / Q000494:pharmacology; D018938:Caco-2 Cells; D015820:Cadherins / Q000378:metabolism; D024722:Cardiac Myosins / Q000378:metabolism; D062506:Claudin-4; D062507:Claudin-5; D057167:Claudins / Q000378:metabolism; D003911:Dextrans / Q000378:metabolism; D017097:Electric Impedance; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D006801:Humans; D020382:Interleukin-18 / Q000494:pharmacology*; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D018994:Myosin Light Chains / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects; D009570:Nitriles / Q000494:pharmacology; D062793:Occludin; D010539:Permeability / Q000187:drug effects; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D013450:Sulfones / Q000494:pharmacology; D057705:Transendothelial and Transepithelial Migration / Q000187:drug effects / Q000502:physiology*; D062826:Zonula Occludens-1 Protein
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D001381:Azepines; C568157:CLDN4 protein, human; C107466:CLDN5 protein, human; D015820:Cadherins; D062506:Claudin-4; D062507:Claudin-5; D057167:Claudins; D003911:Dextrans; D020382:Interleukin-18; D008565:Membrane Proteins; D018994:Myosin Light Chains; D016328:NF-kappa B; D009570:Nitriles; C568985:OCLN protein, human; D062793:Occludin; D010750:Phosphoproteins; D013450:Sulfones; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; C015219:fluorescein isothiocyanate dextran; C107184:myosin light chain 2; C056218:ML 9; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009219:Myosin-Light-Chain Kinase; D024722:Cardiac Myosins; D016650:Fluorescein-5-isothiocyanate