title: Reactive oxygen species rapidly increase endothelial ICAM-1 ability to bind neutrophils without detectable upregulation.
abstract: We compared the effects of phorbol 12-myristate 13-acetate (PMA) and thrombin with those of nonlytic concentrations of reactive oxygen species (ROS) generated by hypoxanthine (HX)-xanthine oxidase (XO) on the adhesion properties of human umbilical cord vein endothelial cells (HUVEC) to resting polymorphonuclear neutrophils (PMN). PMN adherence to HX-XO-treated HUVEC was increased approximately twofold to 2.5-fold relative to untreated HUVEC, both immediately and after 2 hours. It was not additive to that induced by PMA or thrombin stimulation of HUVEC. ROS-induced adherence was not due to platelet-activating factor (PAF) or P-selectin expression, as it was neither antagonized by BN52021 (PAF receptor antagonist) nor inhibited by anti-P-selectin monoclonal antibody (MoAb), contrary to the increased adhesion of PMA- and thrombin-stimulated HUVEC. PMN preincubated with mannose-6-P or N-acetylneuraminic acid (sialic acid), but not mannose or galactose-6-P, showed reduced adherence to ROS-treated HUVEC, suggesting that carbohydrate molecules were expressed on the latter and served as the ligand for the PMN L-selectin. Intercellular adhesion molecule (ICAM-1), constitutively present on the surface of resting HUVEC, was involved in the PMN adherence to ROS-treated HUVEC, since this adherence was inhibited by anti-ICAM-1, anti-CD11a, anti-CD11b, and anti-CD18 MoAbs. A non-CD18, non-ICAM-1-dependent mechanism is also involved in this adherence, since effects of these MoAbs were not additive; moreover, combinations of anti-CD18 and anti-ICAM-1 MoAbs with mannose-6-P and sialic acid completely inhibited PMN adherence. The increased binding of PMN to HX-XO-exposed HUVEC observed here involved IC-AM-1, but was independent of its upregulation, and another non-ICAM-1-dependent mechanism, in which carbohydrates expressed on HUVEC recognize L-selectin on PMN.
authors.affiliation: INSERM U294, CHU Xavier Bichat, Université Paris, France.
authors.forename: H
authors.initials: H
authors.lastname: Sellak
authors.forename: E
authors.initials: E
authors.lastname: Franzini
authors.forename: J
authors.initials: J
authors.lastname: Hakim
authors.forename: C
authors.initials: C
authors.lastname: Pasquier
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD / Q000276:immunology / Q000502:physiology; D018821:CD18 Antigens; D002374:Catalase / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000276:immunology / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1; D019041:L-Selectin; D008360:Mannosephosphates / Q000494:pharmacology; D019158:N-Acetylneuraminic Acid; D009504:Neutrophils / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism / Q000494:pharmacology*; D012794:Sialic Acids / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology; D014471:Umbilical Veins / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018821:CD18 Antigens; D015815:Cell Adhesion Molecules; D007042:Hypoxanthines; D008360:Mannosephosphates; D017382:Reactive Oxygen Species; D012794:Sialic Acids; D018799:Intercellular Adhesion Molecule-1; D019041:L-Selectin; D019271:Hypoxanthine; C027693:mannose-6-phosphate; D002374:Catalase; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D013917:Thrombin; D019158:N-Acetylneuraminic Acid; D013755:Tetradecanoylphorbol Acetate