title: CD11b/CD18-dependent interactions of neutrophils with intestinal epithelium are mediated by fucosylated proteoglycans.
abstract: CD11b/CD18-mediated adhesive interactions play a key role in regulating polymorphonuclear leukocytes (PMN)) migration across intestinal epithelium. However, the identity of epithelial ligands for migrating PMN remains obscure. In this study we investigated the role of carbohydrates in mediating adhesive interactions between T84 intestinal epithelial cells and CD11b/CD18 purified from PMN. Fucoidin, heparin/heparin sulfate, N-acetyl-D-glucosamine, mannose-6-phosphate, and laminarin were found to inhibit adhesion of T84 cells to CD11b/CD18. The most potent inhibitory effects were observed with fucoidin (50% inhibition at 1-5 x 10(-8) M). Binding assays demonstrated that fucoidin directly bound to CD11b/CD18 in a divalent cation- and sulfation-dependent fashion that was blocked by anti-CD11b mAbs. Experiments employing CD11b/CD18 as a probe to blot T84 cell fucosylated proteins purified via fucose-specific lectin column revealed several candidate CD11b/CD18 binding proteins with molecular masses of 95, 50, 30, 25, and 20 kDa. Fucosidase treatment of T84 cells resulted in significantly reduced cell adhesion to CD11b/CD18, while no inhibition was observed after neuraminidase treatment. Finally, significant inhibition of T84 cell adhesion to CD11b/CD18 was observed after blocking cell proteoglycan synthesis with p-nitrophenyl-beta-D-xylopyranoside. These findings implicate epithelial cell surface proteoglycans decorated with sulfated fucose moieties as ligands for CD11b/CD18 during PMN migration across mucosal surfaces.
authors.affiliation: Division of Gastrointestinal Pathology, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322, USA. kzen@emory.edu
authors.forename: Ke
authors.initials: K
authors.lastname: Zen
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
authors.forename: Dana
authors.initials: D
authors.lastname: Cairo
authors.forename: Charles A
authors.initials: CA
authors.lastname: Parkos
mesh_terms: D001667:Binding, Competitive / Q000276:immunology; D019921:Biotinylation; D039481:CD11b Antigen / Q000378:metabolism / Q000502:physiology*; D018821:CD18 Antigens / Q000378:metabolism / Q000502:physiology*; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D005643:Fucose / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism* / Q000502:physiology; D011134:Polysaccharides / Q000378:metabolism; D011485:Protein Binding / Q000276:immunology; D011506:Proteins / Q000378:metabolism; D011509:Proteoglycans / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism / Q000502:physiology*; D013431:Sulfates / Q000378:metabolism; D014407:Tumor Cells, Cultured; D005644:alpha-L-Fucosidase / Q000494:pharmacology
chemical_list: D039481:CD11b Antigen; D018821:CD18 Antigens; D002241:Carbohydrates; D011134:Polysaccharides; D011506:Proteins; D011509:Proteoglycans; D013431:Sulfates; D005643:Fucose; C007789:fucoidan; D005644:alpha-L-Fucosidase