title: Increased expression of bioactive chemokines in human cerebromicrovascular endothelial cells and astrocytes subjected to simulated ischemia in vitro.
abstract: Leukocyte infiltration into the brain has been implicated in the development of ischemic brain damage. In this study, simulated in vitro ischemia/reperfusion and IL-1beta were found to up-regulate both the expression of intercellular adhesion molecule- (ICAM-1) in cultured human cerebromicrovascular endothelial cells (HCEC) and the adhesion of allogenic neutrophils to HCEC. Both HCEC and human fetal astrocytes (FHAS) also responded to IL-1beta and to in vitro ischemia/reperfusion by a pronounced up-regulation of IL-8 and MCP-1 mRNA and by increased release of IL-8 and MCP-1 in cell culture media. FHAS were found to release 30-times higher levels of MCP-1 than HCEC under both basal and ischemic conditions. However, 100 u/ml IL-1beta induced greater stimulation of both IL-8 and MCP-1 secretion in HCEC (50 and 20 times above controls, respectively) than in FHAS (three and two times above controls, respectively). IL-8 was the principal neutrophil chemoattractant released from IL-1beta-treated HCEC, since IL-8 antibody completely inhibited neutrophil chemotaxis enticed by HCEC media. However, the IL-8 antibody neutralized only 50% of IL-1beta-stimulated neutrophil chemoattractants released from FHAS, and 40%-60% of ischemia-stimulated chemotactic activity released by either HCEC or FHAS. These results suggest that simulated in vitro ischemia, in addition to IL-8 and MCP-1, stimulates secretion of other bioactive chemokines from HCEC and FHAS.
authors.affiliation: Institute for Biological Sciences, National Research Council of Canada, Ottawa.
authors.forename: W
authors.initials: W
authors.lastname: Zhang
authors.forename: C
authors.initials: C
authors.lastname: Smith
authors.forename: A
authors.initials: A
authors.lastname: Shapiro
authors.forename: R
authors.initials: R
authors.lastname: Monette
authors.forename: J
authors.initials: J
authors.lastname: Hutchison
authors.forename: D
authors.initials: D
authors.lastname: Stanimirovic
mesh_terms: D001253:Astrocytes / Q000378:metabolism*; D001812:Blood-Brain Barrier; D001921:Brain / Q000098:blood supply*; D002545:Brain Ischemia / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000096:biosynthesis; D018925:Chemokines / Q000096:biosynthesis*; D002634:Chemotaxis, Leukocyte; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D007375:Interleukin-1 / Q000494:pharmacology; D016209:Interleukin-8 / Q000096:biosynthesis; D009504:Neutrophils / Q000276:immunology
chemical_list: D018932:Chemokine CCL2; D018925:Chemokines; D007375:Interleukin-1; D016209:Interleukin-8; D018799:Intercellular Adhesion Molecule-1