Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1995 Apr;95(4):1756–1765. doi: 10.1172/JCI117853

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

F Díaz-González 1, I González-Alvaro 1, M R Campanero 1, F Mollinedo 1, M A del Pozo 1, C Muñoz 1, J P Pivel 1, F Sánchez-Madrid 1
PMCID: PMC295697  PMID: 7535797

Abstract

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.

Full text

PDF
1756

Images in this article

1758Image
on p.1758
1759Image
on p.1759
1759Image
on p.1759
1759Image
on p.1759
1764Image
on p.1764
1764Image
on p.1764

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abramson S. B., Weissmann G. The mechanisms of action of nonsteroidal antiinflammatory drugs. Arthritis Rheum. 1989 Jan;32(1):1–9. doi: 10.1002/anr.1780320102. [DOI] [PubMed] [Google Scholar]
  2. Baumheter S., Singer M. S., Henzel W., Hemmerich S., Renz M., Rosen S. D., Lasky L. A. Binding of L-selectin to the vascular sialomucin CD34. Science. 1993 Oct 15;262(5132):436–438. doi: 10.1126/science.7692600. [DOI] [PubMed] [Google Scholar]
  3. Berg E. L., McEvoy L. M., Berlin C., Bargatze R. F., Butcher E. C. L-selectin-mediated lymphocyte rolling on MAdCAM-1. Nature. 1993 Dec 16;366(6456):695–698. doi: 10.1038/366695a0. [DOI] [PubMed] [Google Scholar]
  4. Berg M., James S. P. Human neutrophils release the Leu-8 lymph node homing receptor during cell activation. Blood. 1990 Dec 1;76(11):2381–2388. [PubMed] [Google Scholar]
  5. Bevilacqua M. P. Endothelial-leukocyte adhesion molecules. Annu Rev Immunol. 1993;11:767–804. doi: 10.1146/annurev.iy.11.040193.004003. [DOI] [PubMed] [Google Scholar]
  6. Brooks P. M., Day R. O. Nonsteroidal antiinflammatory drugs--differences and similarities. N Engl J Med. 1991 Jun 13;324(24):1716–1725. doi: 10.1056/NEJM199106133242407. [DOI] [PubMed] [Google Scholar]
  7. Butcher E. C. Leukocyte-endothelial cell recognition: three (or more) steps to specificity and diversity. Cell. 1991 Dec 20;67(6):1033–1036. doi: 10.1016/0092-8674(91)90279-8. [DOI] [PubMed] [Google Scholar]
  8. Campanero M. R., Pulido R., Alonso J. L., Pivel J. P., Pimentel-Muiños F. X., Fresno M., Sánchez-Madrid F. Down-regulation by tumor necrosis factor-alpha of neutrophil cell surface expression of the sialophorin CD43 and the hyaluronate receptor CD44 through a proteolytic mechanism. Eur J Immunol. 1991 Dec;21(12):3045–3048. doi: 10.1002/eji.1830211222. [DOI] [PubMed] [Google Scholar]
  9. Campanero M. R., del Pozo M. A., Arroyo A. G., Sánchez-Mateos P., Hernández-Caselles T., Craig A., Pulido R., Sánchez-Madrid F. ICAM-3 interacts with LFA-1 and regulates the LFA-1/ICAM-1 cell adhesion pathway. J Cell Biol. 1993 Nov;123(4):1007–1016. doi: 10.1083/jcb.123.4.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Forrest M., Brooks P. M. Mechanism of action of non-steroidal anti-rheumatic drugs. Baillieres Clin Rheumatol. 1988 Aug;2(2):275–294. doi: 10.1016/s0950-3579(88)80015-3. [DOI] [PubMed] [Google Scholar]
  11. Griffin J. D., Spertini O., Ernst T. J., Belvin M. P., Levine H. B., Kanakura Y., Tedder T. F. Granulocyte-macrophage colony-stimulating factor and other cytokines regulate surface expression of the leukocyte adhesion molecule-1 on human neutrophils, monocytes, and their precursors. J Immunol. 1990 Jul 15;145(2):576–584. [PubMed] [Google Scholar]
  12. Humbría A., Díaz-González F., Campanero M. R., Arroyo A. G., Laffón A., González-Amaro R., Sánchez-Madrid F. Expression of L-selectin, CD43, and CD44 in synovial fluid neutrophils from patients with inflammatory joint diseases. Evidence for a soluble form of L-selectin in synovial fluid. Arthritis Rheum. 1994 Mar;37(3):342–348. doi: 10.1002/art.1780370307. [DOI] [PubMed] [Google Scholar]
  13. Imai Y., Singer M. S., Fennie C., Lasky L. A., Rosen S. D. Identification of a carbohydrate-based endothelial ligand for a lymphocyte homing receptor. J Cell Biol. 1991 Jun;113(5):1213–1221. doi: 10.1083/jcb.113.5.1213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jutila M. A., Rott L., Berg E. L., Butcher E. C. Function and regulation of the neutrophil MEL-14 antigen in vivo: comparison with LFA-1 and MAC-1. J Immunol. 1989 Nov 15;143(10):3318–3324. [PubMed] [Google Scholar]
  15. Kishimoto T. K., Jutila M. A., Berg E. L., Butcher E. C. Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors. Science. 1989 Sep 15;245(4923):1238–1241. doi: 10.1126/science.2551036. [DOI] [PubMed] [Google Scholar]
  16. Kopp E., Ghosh S. Inhibition of NF-kappa B by sodium salicylate and aspirin. Science. 1994 Aug 12;265(5174):956–959. doi: 10.1126/science.8052854. [DOI] [PubMed] [Google Scholar]
  17. Lacal P., Pulido R., Sánchez-Madrid F., Mollinedo F. Intracellular location of T200 and Mo1 glycoproteins in human neutrophils. J Biol Chem. 1988 Jul 15;263(20):9946–9951. [PubMed] [Google Scholar]
  18. Langman M. J. Ulcer complications and nonsteroidal anti-inflammatory drugs. Am J Med. 1988 Feb 22;84(2A):15–19. doi: 10.1016/0002-9343(88)90249-5. [DOI] [PubMed] [Google Scholar]
  19. Lasky L. A. Selectins: interpreters of cell-specific carbohydrate information during inflammation. Science. 1992 Nov 6;258(5084):964–969. doi: 10.1126/science.1439808. [DOI] [PubMed] [Google Scholar]
  20. Lasky L. A., Singer M. S., Dowbenko D., Imai Y., Henzel W. J., Grimley C., Fennie C., Gillett N., Watson S. R., Rosen S. D. An endothelial ligand for L-selectin is a novel mucin-like molecule. Cell. 1992 Jun 12;69(6):927–938. doi: 10.1016/0092-8674(92)90612-g. [DOI] [PubMed] [Google Scholar]
  21. Lawrence M. B., Springer T. A. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991 May 31;65(5):859–873. doi: 10.1016/0092-8674(91)90393-d. [DOI] [PubMed] [Google Scholar]
  22. Ley K., Gaehtgens P., Fennie C., Singer M. S., Lasky L. A., Rosen S. D. Lectin-like cell adhesion molecule 1 mediates leukocyte rolling in mesenteric venules in vivo. Blood. 1991 Jun 15;77(12):2553–2555. [PubMed] [Google Scholar]
  23. McEver R. P., Beckstead J. H., Moore K. L., Marshall-Carlson L., Bainton D. F. GMP-140, a platelet alpha-granule membrane protein, is also synthesized by vascular endothelial cells and is localized in Weibel-Palade bodies. J Clin Invest. 1989 Jul;84(1):92–99. doi: 10.1172/JCI114175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McEver R. P. Leukocyte-endothelial cell interactions. Curr Opin Cell Biol. 1992 Oct;4(5):840–849. doi: 10.1016/0955-0674(92)90109-p. [DOI] [PubMed] [Google Scholar]
  25. Miller L. J., Bainton D. F., Borregaard N., Springer T. A. Stimulated mobilization of monocyte Mac-1 and p150,95 adhesion proteins from an intracellular vesicular compartment to the cell surface. J Clin Invest. 1987 Aug;80(2):535–544. doi: 10.1172/JCI113102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mollinedo F., Manara F. S., Schneider D. L. Acidification activity of human neutrophils. Tertiary granules as a site of ATP-dependent acidification. J Biol Chem. 1986 Jan 25;261(3):1077–1082. [PubMed] [Google Scholar]
  27. Mulligan M. S., Paulson J. C., De Frees S., Zheng Z. L., Lowe J. B., Ward P. A. Protective effects of oligosaccharides in P-selectin-dependent lung injury. Nature. 1993 Jul 8;364(6433):149–151. doi: 10.1038/364149a0. [DOI] [PubMed] [Google Scholar]
  28. Palecanda A., Walcheck B., Bishop D. K., Jutila M. A. Rapid activation-independent shedding of leukocyte L-selectin induced by cross-linking of the surface antigen. Eur J Immunol. 1992 May;22(5):1279–1286. doi: 10.1002/eji.1830220524. [DOI] [PubMed] [Google Scholar]
  29. Picker L. J., Warnock R. A., Burns A. R., Doerschuk C. M., Berg E. L., Butcher E. C. The neutrophil selectin LECAM-1 presents carbohydrate ligands to the vascular selectins ELAM-1 and GMP-140. Cell. 1991 Sep 6;66(5):921–933. doi: 10.1016/0092-8674(91)90438-5. [DOI] [PubMed] [Google Scholar]
  30. Sanchez-Madrid F., Nagy J. A., Robbins E., Simon P., Springer T. A. A human leukocyte differentiation antigen family with distinct alpha-subunits and a common beta-subunit: the lymphocyte function-associated antigen (LFA-1), the C3bi complement receptor (OKM1/Mac-1), and the p150,95 molecule. J Exp Med. 1983 Dec 1;158(6):1785–1803. doi: 10.1084/jem.158.6.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schleiffenbaum B., Spertini O., Tedder T. F. Soluble L-selectin is present in human plasma at high levels and retains functional activity. J Cell Biol. 1992 Oct;119(1):229–238. doi: 10.1083/jcb.119.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shimizu Y., Newman W., Tanaka Y., Shaw S. Lymphocyte interactions with endothelial cells. Immunol Today. 1992 Mar;13(3):106–112. doi: 10.1016/0167-5699(92)90151-V. [DOI] [PubMed] [Google Scholar]
  33. Spertini O., Kansas G. S., Munro J. M., Griffin J. D., Tedder T. F. Regulation of leukocyte migration by activation of the leukocyte adhesion molecule-1 (LAM-1) selectin. Nature. 1991 Feb 21;349(6311):691–694. doi: 10.1038/349691a0. [DOI] [PubMed] [Google Scholar]
  34. Spertini O., Kansas G. S., Reimann K. A., Mackay C. R., Tedder T. F. Function and evolutionary conservation of distinct epitopes on the leukocyte adhesion molecule-1 (TQ-1, Leu-8) that regulate leukocyte migration. J Immunol. 1991 Aug 1;147(3):942–949. [PubMed] [Google Scholar]
  35. Spertini O., Luscinskas F. W., Gimbrone M. A., Jr, Tedder T. F. Monocyte attachment to activated human vascular endothelium in vitro is mediated by leukocyte adhesion molecule-1 (L-selectin) under nonstatic conditions. J Exp Med. 1992 Jun 1;175(6):1789–1792. doi: 10.1084/jem.175.6.1789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Spertini O., Luscinskas F. W., Kansas G. S., Munro J. M., Griffin J. D., Gimbrone M. A., Jr, Tedder T. F. Leukocyte adhesion molecule-1 (LAM-1, L-selectin) interacts with an inducible endothelial cell ligand to support leukocyte adhesion. J Immunol. 1991 Oct 15;147(8):2565–2573. [PubMed] [Google Scholar]
  37. Springer T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994 Jan 28;76(2):301–314. doi: 10.1016/0092-8674(94)90337-9. [DOI] [PubMed] [Google Scholar]
  38. Stenberg P. E., McEver R. P., Shuman M. A., Jacques Y. V., Bainton D. F. A platelet alpha-granule membrane protein (GMP-140) is expressed on the plasma membrane after activation. J Cell Biol. 1985 Sep;101(3):880–886. doi: 10.1083/jcb.101.3.880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tedder T. F., Matsuyama T., Rothstein D., Schlossman S. F., Morimoto C. Human antigen-specific memory T cells express the homing receptor (LAM-1) necessary for lymphocyte recirculation. Eur J Immunol. 1990 Jun;20(6):1351–1355. doi: 10.1002/eji.1830200622. [DOI] [PubMed] [Google Scholar]
  40. Travis J. Biotech gets a grip on cell adhesion. Science. 1993 May 14;260(5110):906–908. doi: 10.1126/science.8493523. [DOI] [PubMed] [Google Scholar]
  41. True D. D., Singer M. S., Lasky L. A., Rosen S. D. Requirement for sialic acid on the endothelial ligand of a lymphocyte homing receptor. J Cell Biol. 1990 Dec;111(6 Pt 1):2757–2764. doi: 10.1083/jcb.111.6.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Vane J. R. Inhibition of prostaglandin synthesis as a mechanism of action for aspirin-like drugs. Nat New Biol. 1971 Jun 23;231(25):232–235. doi: 10.1038/newbio231232a0. [DOI] [PubMed] [Google Scholar]
  43. Watson S. R., Fennie C., Lasky L. A. Neutrophil influx into an inflammatory site inhibited by a soluble homing receptor-IgG chimaera. Nature. 1991 Jan 10;349(6305):164–167. doi: 10.1038/349164a0. [DOI] [PubMed] [Google Scholar]
  44. Yednock T. A., Stoolman L. M., Rosen S. D. Phosphomannosyl-derivatized beads detect a receptor involved in lymphocyte homing. J Cell Biol. 1987 Mar;104(3):713–723. doi: 10.1083/jcb.104.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zimmerman G. A., Prescott S. M., McIntyre T. M. Endothelial cell interactions with granulocytes: tethering and signaling molecules. Immunol Today. 1992 Mar;13(3):93–100. doi: 10.1016/0167-5699(92)90149-2. [DOI] [PubMed] [Google Scholar]
  46. von Andrian U. H., Chambers J. D., McEvoy L. M., Bargatze R. F., Arfors K. E., Butcher E. C. Two-step model of leukocyte-endothelial cell interaction in inflammation: distinct roles for LECAM-1 and the leukocyte beta 2 integrins in vivo. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7538–7542. doi: 10.1073/pnas.88.17.7538. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES