title: Mechanism of cigarette smoke condensate induced adhesion of human monocytes to cultured endothelial cells.
abstract: Cigarette smoking is ranked among the leading risk factors in the etiology of atherosclerotic vascular disease. The mechanisms, however, that link cigarette smoking to increased incidence of atherosclerosis are not understood. The adherence of circulating monocytes to the endothelium, migration into the subendothelium, and subsequent formation of foam cells are principal initial events in the development of atherosclerosis. We therefore determined whether cigarette smoke caused increased adherence of monocytes to endothelial cells and the cellular mechanism of this increased adherence. Cigarette smoke condensate (CSC), the particulate fraction of cigarette smoke derived from 2R1 standard research cigarettes, at a concentration of 25-30 micrograms/ml (average yield of CSC is 26.1 mg/cigarette), augmented (70-90%) basal adherence of human peripheral blood monocytes to a cultured monolayer of endothelial cells derived from bovine aorta (BAEC) and human umbilical vein (HUVEC). There was a concomitant increase in the expression of CD11b ligand on the surface of monocytes as determined by flow cytometry, utilizing FITC conjugated Mab MO-1 (CD11b). However, nicotine (1-15 micrograms/ml) and cadmium sulfate (10 micrograms/ml), constituents of CSC, individually or in combination had no effect either on CD11b expression or adherence of monocytes to endothelial cells. Treatment of HUVEC with CSC for 60 min also resulted in an increased expression of ICAM-1 and ELAM-1 as determined by mean fluorescence intensity of ICAM-1 and ELAM-1 labeled cells in flow cytometric analysis. The CSC induced expression of CD11b in monocytes was optimal at 25-30 min and was inhibited by protein kinase C inhibitors, staurosporine and H-7, and also by baicalein, a lipoxygenase inhibitor. Similarly, CSC induced ICAM-1 and ELAM-1 expression in HUVEC was inhibited by protein kinase C inhibitors. CSC stimulated the adherence of human monocytes but not the monocytic cell lines HL-60, U937, and THP-1 to endothelial cells. The CSC stimulated adherence of human monocytes was inhibited (80%) by MAb to CD11b and 50% by Mab to ICAM-1 and ELAM-1. These results suggest that cigarette smoke particulate constituents activate protein kinase C, leading to increased surface expression of adhesive ligand CD11b on peripheral blood monocytes and counter receptor(s) ICAM-1 and ELAM-1 in endothelial cells. The expression of ligand and counter receptor leads to potentiated adherence of monocytes to endothelial cells, an initial event in the pathogenesis of cigarette smoke induced inflammatory response in the vessel wall.
authors.affiliation: Department of Biochemistry, School of Medicine, Children's Hospital of Los Angeles, University of Southern California 90033.
authors.forename: V K
authors.initials: VK
authors.lastname: Kalra
authors.forename: Y
authors.initials: Y
authors.lastname: Ying
authors.forename: K
authors.initials: K
authors.lastname: Deemer
authors.forename: R
authors.initials: R
authors.lastname: Natarajan
authors.forename: J L
authors.initials: JL
authors.lastname: Nadler
authors.forename: T D
authors.initials: TD
authors.lastname: Coates
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D015703:Antigens, CD / Q000032:analysis / Q000502:physiology; D018845:CD11 Antigens; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D005434:Flow Cytometry; D006801:Humans; D018799:Intercellular Adhesion Molecule-1; D009000:Monocytes / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D009538:Nicotine / Q000494:pharmacology; D010947:Plants, Toxic; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology; D012906:Smoke; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014026:Nicotiana; D019010:Vascular Cell Adhesion Molecule-1
chemical_list: D000470:Alkaloids; D015703:Antigens, CD; D018845:CD11 Antigens; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D012906:Smoke; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D009538:Nicotine; D011493:Protein Kinase C; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate