title: Generation of neutrophil chemotactic activity by phorbol ester-stimulated calf pulmonary artery endothelial cells.
abstract: Chemotactic activity for human polymorphonuclear leukocytes (PMNL) was detected in serum-free conditioned media 1 to 4 hr after monolayers of calf pulmonary artery endothelial cells were pretreated with phorbol myristate acetate (PMA). Chemotactic activity was increased in conditioned media following pretreatment with either PMA or the less lipophilic active phorbol ester, 4-beta-phorbol-12,13-dibutyrate (P(Bu)2) in a dose-dependent manner. Chemotactic activity of conditioned media from PMA-treated endothelial cells was confirmed by checkerboard analysis. The chemotactic activity in conditioned media from PMA-pretreated endothelial cells was completely inhibited by pretreating endothelial cells with either cycloheximide, actinomycin D, or the lipooxygenase inhibitor, diethylcarbamazine. Furthermore, the chemotactic activity was heat-stable, inhibited by trypsin treatment, and present in both aqueous and lipid phases after ether extraction. The data demonstrate that pulmonary artery endothelial cells exposed to active phorbol esters release potent chemotactic factor(s) for PMNL. These findings suggest a role for activators of protein kinase C in mediating endothelial cell release of chemotactic factor(s) that may be important in the directed migration of circulating leukocytes to sites of vascular injury.
authors.affiliation: Department of Physiology, Albany Medical College, Union University, NY 12208.
authors.forename: P W
authors.initials: PW
authors.lastname: Gudewicz
authors.forename: L E
authors.initials: LE
authors.lastname: Odekon
authors.forename: P J
authors.initials: PJ
authors.lastname: Del Vecchio
authors.forename: T M
authors.initials: TM
authors.lastname: Saba
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002630:Chemotactic Factors / Q000096:biosynthesis*; D003470:Culture Media; D003609:Dactinomycin / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism*; D016209:Interleukin-8; D009504:Neutrophils / Q000276:immunology*; D015240:Phorbol 12,13-Dibutyrate; D010703:Phorbol Esters / Q000494:pharmacology*; D011651:Pulmonary Artery; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
chemical_list: D002630:Chemotactic Factors; D003470:Culture Media; D016209:Interleukin-8; D010703:Phorbol Esters; D003609:Dactinomycin; D015240:Phorbol 12,13-Dibutyrate; D013755:Tetradecanoylphorbol Acetate