Prevention of in vitro neutrophil-endothelial attachment through shedding of L-selectin by nonsteroidal antiinflammatory drugs

• Published in: The Journal of clinical investigation Vol. 95; no. 4; pp. 1756 - 1765
• Main Authors: Díaz-González, F, González-Alvaro, I, Campanero, M R, Mollinedo, F, del Pozo, M A, Muñoz, C, Pivel, J P, Sánchez-Madrid, F
• Format: Journal Article
• Language: English
• Published: United States 01.04.1995
• Subjects: Anti-Inflammatory Agents, Non-Steroidal - pharmacology; Antigens, CD - biosynthesis; Aspirin - pharmacology; Cell Adhesion - drug effects; Cell Adhesion Molecules - metabolism; Cytoplasmic Granules - enzymology; Diclofenac - pharmacology; Dose-Response Relationship, Drug; Down-Regulation; Endothelium, Vascular - physiology; Enzymes - secretion; Humans; Indomethacin - pharmacology; Inflammation - etiology; Intercellular Adhesion Molecule-1 - biosynthesis; L-Selectin; Neutrophils - physiology; Piroxicam - pharmacology
• DOI: 10.1172/jci117853
• ISSN: 0021-9738

The activation of the endothelial cells by extravascular stimuli is the key event in the extravasation of circulating leukocytes to target tissues. L-selectin, a member of the selectin family, is constitutively expressed by white cells, and is the molecule involved in the initial binding of leukocytes to activated endothelium. After activation, leukocytes rapidly release L-selectin from the cell surface, suggesting that the functional activity of this molecule is controlled in large part by its appearance and disappearance from cell surface. We have studied in a neutrophil-activated endothelial cell binding assay, the effect of different antiinflammatory drugs (steroidal and nonsteroidal) in the L-selectin-mediated interaction of neutrophils with activated endothelial cells. Some nonsteroidal antiinflammatory drugs (NSAIDs), such as indomethacin, diclofenac, ketoprofen, and aspirin, but not steroids, strongly inhibited the neutrophil-endothelial cell attachment. Furthermore, we also investigated the underlying mechanism of this functional effect. The expression of L-selectin on the neutrophil surface rapidly decreased in the presence of different NSAIDs, in a dose- and time-dependent manner, whereas no changes in the expression of other adhesion molecules such as CD11a, CD11b, CD31, or ICAM-3 (CD50) were observed. Interestingly, studies in vivo on healthy volunteers treated with physiological doses of indomethacin showed a significant decrease of L-selectin neutrophil expression. Only diclofenac induced an upregulation of CD11b expression, suggesting an activating effect on neutrophils. No enzyme release was observed upon treatment of neutrophils with different NSAIDs, indicating a lack of degranulatory activity of NSAIDs, with the exception of diclofenac. The downregulation of L-selectin expression was due to the rapid cleavage and shedding of the membrane L-selectin, as determined by both immunoprecipitation from 125I-labeled neutrophils, and quantitative estimation in cell-free supernatants. These results suggest that NSAIDs exert a specific action on adhesion receptor expression in neutrophils, which might account, at least in part, for the antiinflammatory activities of NSAIDs.