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. Copyright 2001 Academic Press.