Stymulacja czasteczek adhezyjnych srodblonka naczyniowego przez polisacharyd otoczkowy, lipopolisacharyd oraz komponenty lipopolisacharydu Bacteroides thetaiotaomicron

Badano wpływ antygenów powierzchniowych: polisacharydu otoczkowego, lipopolisacharydu i komponentów LPS B. thetaiotaomicron na ekspresję molekuł adhezyjnych ICAM-1, VCAM-1 i selektyny E na powierzchni komórek śródbłonka naczyniowego (linia HMEC-1). Cząsteczki adhezyjne oznaczano za pomocą testu immunoenzymatycznego ELISA z przeciwciałami monoklonalnymi dla ludzkich: ICAM-1, VCAM- 1 i selektyny E. Najsilniejszymi stymulatorami cząsteczek adhezyjnych były lipopolisacharydy, a następnie części lipidowe LPS (lipid A). Polisacharyd otoczkowy (CPS) stymulował ekspresję wszystkich molekuł adhezyjnych, ale tylko w największym stężeniu. Nąjsłabszymi stymulatorami ICAM-1, VCAM-1 i selektyny E były części polisacharydowe (PS) LPS. Niektóre preparaty PS nie wykazały aktywności stymulacyjnej.

The aim of this study was to assay the influence of capsular polysaccharide (CPS), lipopolysaccharide (LPS) and components of B. thetaiotaomicron lipopolysaccharide - polysaccharide part (PS) and lipid part (lipid A) on the expression of adhesion molecules associated with inflammation (ICAM-1, VCAM-1, E-selectin) on the surface of vascular endothelial cells. Capsular polysaccharide was isolated by the method of Poxton and Ip (1981). Lipopolysaccharides were extracted using the hot phenol-water method (Westphal and Jann, 1965). Components of LPS were prepared by mild acid hydrolysis of lipopolysaccharide. Experiments with bacterial compounds at concentrations 10, 1, 0.1 and 0.01 (mg/ml) were performed on HMEC-1 cell line (human dermal microvascular endothelial cells). Immunoenzymatic ELISA test with mouse monoclonal antibodies against human: ICAM-1, VCAM-1 and E-selectin was applied to determine adhesion molecules. Resting HMEC-1 and E. coli 055:B5 LPS were used as controls in each experiment. Lipopolysaccharides were the strongest stimulants of endothelial adhesion molecules. Capsular polysaccharide caused the expression of three adhesion molecules, but only at the highest concentration (10 mg/ml). The stimulatory activities of LPS lipid components were much higher than the activities of polysaccharide parts. PS preparations did not reveal the property of adhesion molecule stimulation or their activities were weak. The activity of B. thetaiotaomicron cell-surface antigens in the process of adhesion molecule stimulation on vascular endothelium was lower than the activity of E. coli LPS.