The kinetics, function, and regulation of P-selectin expressed by human brain microvessel endothelial cells in primary culture

P-selectin is an endothelial cell adhesion glycoprotein expressed on the cell surface early in inflammation where it binds to blood leukocytes. This study examines the expression, function, and regulation of P-selectin in primary cultures of human brain microvessel endothelial cells (HBMEC). Surface expression of P-selectin was minimal in unstimulated HBMEC; however, it was significantly augmented upon stimulation with histamine (10^-7-10^-3 M) and thrombin (0.01-1 U/ml). Expression increased rapidly at 10 min and remained elevated at 60 min. Immunogold electron microscopy showed that histamine (10^-7 M) increased surface expression preferentially on the apical surface of subconfluent monolayers. A cell binding assay showed that the adhesion of polymorphonuclear leukocytes (PMNs) to confluent monolayers was augmented after histamine treatment. Histamine-induced surface expression of P-selectin was blocked by the histamine H₂ receptor antagonist cimetidine. The H₁ receptor antagonist mepyramine had no effect. Expression was reduced by the extracellular calcium chelator EDTA and blocked by the cyclic AMP phosphodiesterase inhibitor rolipram. Thus histamine and thrombin both increase P-selectin expression in HBMEC. Histamine mediates expression through the H₂, but not the H₁, receptor and calcium, whereas expression is reduced by cyclic AMP. The histamine-induced expression increases PMN binding to the HBMEC. These data suggest that P-selectin plays a role in the recruitment of acute inflammatory cells to the CNS.