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Human neutrophil adhesion to bovine aortic endothelium. Evidence for endothelial lipoxygenase activity
Author(s) -
Damtew Belai,
Goldsmith George,
Tserng KoyYi,
Spagnuolo Philip J.
Publication year - 1993
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.53.6.619
Subject(s) - leukotriene b4 , endothelium , phorbol , nordihydroguaiaretic acid , biology , endothelial stem cell , bradykinin , thrombin , thromboxane a2 , endocrinology , biochemistry , medicine , microbiology and biotechnology , protein kinase c , arachidonic acid , immunology , in vitro , platelet , enzyme , inflammation , receptor
We examined the effect of phorbol myristate acetate on cultured bovine aortic endothelial cells to determine the role of endothelial cells in neutrophil‐endothelial cell adhesive interactions. Confluent endothelial cells were preincubated with phorbol myristate acetate and other inflammatory signals including JV‐formylmethionyl‐ leucyl‐phenylalanine (f‐Met‐Leu‐Phe), the ionophore A23187, and thrombin; washed extensively; and incubated with 51 Cr‐labeled neutrophils. Preincubation of endothelium with A23187, phorbol ester, or thrombin increased adherence of neutrophils by 3.1‐, 5.7‐, and 3.7‐fold over baseline. In contrast, f‐Met‐Leu‐Phe preincubation failed to increase adhesion over baseline. Supernatants from endothelium preincubated with phorbol failed to augment adherence of untreated endothelial cells. Preincubation of endothelium with lipoxygenase inhibitors nordihydroguaiaretic acid (50μM), 5,8,11,14‐eicosatetraenoic acid (50 μ M), and BW755C (50 μ M) inhibited the effect of phorbol preincubation of endothelium significantly by 55, 27, and 22%, respectively. In contrast, inhibitors of cyclooxygenase and thromboxane synthase or thromboxane receptor antagonists had no effect on phorbol‐induced adhesion. Specific desensitization of neutrophil adhesion to phorbol‐treated endothelium could be demonstrated by prior exposure of neutrophils to low concentrations of leukotriene B 4 (3.8 × 10 −10 M). Endothelium preincubated with phorbol but not f‐Met‐Leu‐Phe or thrombin produced several fatty acid peaks at 280 nm, one of which comigrated with authentic leukotriene B 4 (LTB 4 ). This peak, isolated and purified, increased endothelial cell adherence in a temporal fashion in the same way as LTB 4 and was demonstrated to be LTB 4 by ultraviolet spectroscopy, high‐ performance liquid chromatography, and mass spectroscopy. These data demonstrate that endothelial cell‐derived lipoxygenase metabolites, in particular LTB 4 , are involved, in part, in the acute regulation of neutrophil adhesion to endothelium induced by inflammatory signals such as phorbol ester.