Human neutrophil adhesion to bovine aortic endothelium. Evidence for endothelial lipoxygenase activity

• Published in: Journal of leukocyte biology Vol. 53; no. 6; pp. 619 - 629
• Main Authors: Damtew, Belai, Goldsmith, George, Tserng, Koy‐Yi, Spagnuolo, Philip J.
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Society for Leukocyte Biology 01.06.1993; Federation of American Societies for Experimental Biology
• Subjects: Animals; Arachidonic Acid - metabolism; Biological and medical sciences; Cattle; Cell Adhesion; Cell interactions, adhesion; Cells, Cultured; endothelium; Endothelium, Vascular - enzymology; Endothelium, Vascular - physiology; Fundamental and applied biological sciences. Psychology; Humans; Inflammation - pathology; Leukotriene B4 - physiology; lipoxygenase; Lipoxygenase - metabolism; Molecular and cellular biology; N-Formylmethionine Leucyl-Phenylalanine - pharmacology; neutrophil; Neutrophils - physiology; Tetradecanoylphorbol Acetate - pharmacology; Thrombin - pharmacology; Human; Endothelial cell; Bovine; Enzyme; Arachidonic acid derivatives; Cell cell interaction; Adhesion; Endothelium; Vertebrata; Mammalia; Enzymatic activity; Leukotriene B4; Aorta; Artiodactyla; Neutrophil; Lipoxygenase; Ungulata
• ISSN: 0741-5400; 1938-3673
• DOI: 10.1002/jlb.53.6.619

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 51Cr‐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 μM), and BW755C (50 μM) 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 B4 (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 B4 (LTB4). This peak, isolated and purified, increased endothelial cell adherence in a temporal fashion in the same way as LTB4 and was demonstrated to be LTB4 by ultraviolet spectroscopy, high‐ performance liquid chromatography, and mass spectroscopy. These data demonstrate that endothelial cell‐derived lipoxygenase metabolites, in particular LTB4, are involved, in part, in the acute regulation of neutrophil adhesion to endothelium induced by inflammatory signals such as phorbol ester.