title: Regulated complement deposition on the surface of human endothelial cells: effect of tobacco smoke and shear stress.
abstract: Cigarette smoke and hemodynamic stress both contribute to vascular inflammation and associated atherosclerosis. We recently demonstrated direct activation of complement components C4 and C3 on human endothelial cells (EC). The present study was designed to explore complement activation on bone marrow microvascular endothelial cells (BMEC) and human umbilical vein endothelial cells (HUVEC) in response to endothelial cell injury by tobacco smoke extract, shear stress, or other known inflammatory and atherogenic mediators, lipopolysaccharide (LPS) and INF-gamma. Following treatment, confluent EC monolayers were exposed to plasma (60 min, 37 degrees C), and cell surface deposition of stable complement derivatives C4d, iC3b and SC5b-9 was measured in situ using an ELISA approach. Consistent with previous results, moderate levels of C4d, iC3b and SC5b-9 deposition were observed on native EC monolayers exposed to human plasma. Tobacco smoke and shear stress enhanced EC C4d deposition. In contrast, LPS and INF-gamma failed to affect EC mediated complement activation, despite evidence of EC activation illustrated by ICAM-1 expression. The combination of tobacco smoke and shear stress nearly doubled EC C4d expression. No increases in iC3b or SC5b-9 were noted, suggesting inhibition of classical and alternative pathway C3 convertase assembly or activity. Indeed, concomitantly increased surface expression of complement regulatory proteins CD35 (CR1) and CD55 was observed following EC exposure to tobacco smoke and shear stress. These results suggest that a balance between complement activation and regulation exists at the EC surface, and may impact vascular injury leading to thrombosis, arteriosclerosis, and atherogenesis.
authors.affiliation: Department of Pathology and Laboratory of Medicine, Weill Medical College of Cornell University, New York, New York, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Yin
authors.forename: Berhane
authors.initials: B
authors.lastname: Ghebrehiwet
authors.forename: Babette
authors.initials: B
authors.lastname: Weksler
authors.forename: Ellinor I B
authors.initials: EI
authors.lastname: Peerschke
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D018958:CD55 Antigens / Q000096:biosynthesis; D003167:Complement Activation; D003165:Complement System Proteins / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006439:Hemodynamics; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D051379:Mice; D008833:Microcirculation / Q000166:cytology; D017463:Receptors, Complement 3b / Q000096:biosynthesis; D012907:Smoking; D013314:Stress, Mechanical; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D018958:CD55 Antigens; D008070:Lipopolysaccharides; D017463:Receptors, Complement 3b; D007371:Interferon-gamma; D003165:Complement System Proteins