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. 1998 Jul 1;102(1):136–144. doi: 10.1172/JCI2592

Modulation of monocyte-endothelial cell interactions by platelet microparticles.

O P Barry 1, D Praticò 1, R C Savani 1, G A FitzGerald 1
PMCID: PMC509075  PMID: 9649567

Abstract

Platelets, activated by various agonists, produce microparticles (MP) from the plasma membrane, which are released into the extracellular space. Although the mechanism of MP formation has been clarified, their biological importance remains ill defined. We have recently shown that platelet-derived MP influence platelet and endothelial cell function. In this study, we have further examined the mechanism of cellular activation by platelet MP. To address the possibility that they may influence monocyte-endothelial interactions, we used an in vitro assay to examine their effects on the adhesion of monocytes to human umbilical vein endothelial cells (HUVEC). Platelet MP increased the adhesion of monocytes to HUVEC in a time- and dose-dependent manner. Maximal adhesion of monocytes to resting HUVEC was observed after 24 h of stimulation with MP. Similar kinetics were observed with U-937 (human promonocytic leukemia) cells, used as a model for the blood-borne monocyte. Maximal adhesion of resting monocytes to MP-stimulated HUVEC was observed after 5 h of stimulation with MP. The EC50s for MP-induced increases in HUVEC, monocyte, and U-937 cell adhesion is 8.74, 43.41, and 10.83 microg/ml of MP protein, respectively. The induction of monocyte-endothelial adhesion was mimicked by arachidonic acid isolated from MP. The observed increased cellular adhesiveness correlated with MP-induced upregulation of cell adhesion molecules. MP-stimulated HUVEC increased intracellular cell adhesion molecule-1 (ICAM-1) but not vascular cell adhesion molecule-1 (VCAM-1), P-, or E-selectin expression. Monocyte and U-937 lymphocyte function-associated antigen-1 (CD11a/CD18) and macrophage antigen-1 (CD11b/ CD18, alpham/beta2) were both upregulated upon MP stimulation, but an increase in p150,95 (CD11c/CD18), very late antigen-1, or ICAM-1 expression was not observed. The functional importance of these changes was demonstrated with blocking antibodies. MP also induced the chemotaxis of U-937 cells in a dose-dependent manner with an EC50 of 4.40 microg/ml of MP protein. Similarly, arachidonic acid isolated from MP mimicked the chemotactic response. A role for PKC was implicated in both adhesion and chemotaxis. GF 109203X, a specific inhibitor of PKC, significantly reduced monocyte-endothelial adhesion, as well as U-937 chemotaxis. The demonstration that platelet MP may modulate important aspects of endothelial and monocyte function provides a novel mechanism by which platelets may interact with such cells in human atherosclerosis and inflammation.

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Selected References

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  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Barry O. P., Pratico D., Lawson J. A., FitzGerald G. A. Transcellular activation of platelets and endothelial cells by bioactive lipids in platelet microparticles. J Clin Invest. 1997 May 1;99(9):2118–2127. doi: 10.1172/JCI119385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beekhuizen H., Blokland I., van Furth R. Cross-linking of CD14 molecules on monocytes results in a CD11/CD18- and ICAM-1-dependent adherence to cytokine-stimulated human endothelial cells. J Immunol. 1993 Feb 1;150(3):950–959. [PubMed] [Google Scholar]
  4. Beekhuizen H., Corsèl-Van Tilburg A. J., Blokland I., Van Furth R. Characterization of the adherence of human monocytes to cytokine-stimulated human macrovascular endothelial cells. Immunology. 1991 Dec;74(4):661–669. [PMC free article] [PubMed] [Google Scholar]
  5. Bevilacqua M. P., Nelson R. M., Mannori G., Cecconi O. Endothelial-leukocyte adhesion molecules in human disease. Annu Rev Med. 1994;45:361–378. doi: 10.1146/annurev.med.45.1.361. [DOI] [PubMed] [Google Scholar]
  6. Boudoulas H. Factors determining and prevention of atherosclerotic plaque rupture. Postgrad Med J. 1994;70 (Suppl 1):S43–S45. [PubMed] [Google Scholar]
  7. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  8. Carlos T. M., Dobrina A., Ross R., Harlan J. M. Multiple receptors on human monocytes are involved in adhesion to cultured human endothelial cells. J Leukoc Biol. 1990 Nov;48(5):451–456. doi: 10.1002/jlb.48.5.451. [DOI] [PubMed] [Google Scholar]
  9. Cavender D. E., Edelbaum D., Welkovich L. Effects of inflammatory cytokines and phorbol esters on the adhesion of U937 cells, a human monocyte-like cell line, to endothelial cell monolayers and extracellular matrix proteins. J Leukoc Biol. 1991 Jun;49(6):566–578. doi: 10.1002/jlb.49.6.566. [DOI] [PubMed] [Google Scholar]
  10. Claesson-Welsh L. Platelet-derived growth factor receptor signals. J Biol Chem. 1994 Dec 23;269(51):32023–32026. [PubMed] [Google Scholar]
  11. Cookson F. B. The origin of foam cells in atherosclerosis. Br J Exp Pathol. 1971 Feb;52(1):62–69. [PMC free article] [PubMed] [Google Scholar]
  12. Davies P. F., Barbee K. A., Lal R., Robotewskyj A., Griem M. L. Hemodynamics and atherogenesis. Endothelial surface dynamics in flow signal transduction. Ann N Y Acad Sci. 1995 Jan 17;748:86–103. [PubMed] [Google Scholar]
  13. Davies P. F., Barbee K. A., Volin M. V., Robotewskyj A., Chen J., Joseph L., Griem M. L., Wernick M. N., Jacobs E., Polacek D. C. Spatial relationships in early signaling events of flow-mediated endothelial mechanotransduction. Annu Rev Physiol. 1997;59:527–549. doi: 10.1146/annurev.physiol.59.1.527. [DOI] [PubMed] [Google Scholar]
  14. Davies P. F., Tripathi S. C. Mechanical stress mechanisms and the cell. An endothelial paradigm. Circ Res. 1993 Feb;72(2):239–245. doi: 10.1161/01.res.72.2.239. [DOI] [PubMed] [Google Scholar]
  15. Derman M. P., Toker A., Hartwig J. H., Spokes K., Falck J. R., Chen C. S., Cantley L. C., Cantley L. G. The lipid products of phosphoinositide 3-kinase increase cell motility through protein kinase C. J Biol Chem. 1997 Mar 7;272(10):6465–6470. doi: 10.1074/jbc.272.10.6465. [DOI] [PubMed] [Google Scholar]
  16. Diamond M. S., Staunton D. E., de Fougerolles A. R., Stacker S. A., Garcia-Aguilar J., Hibbs M. L., Springer T. A. ICAM-1 (CD54): a counter-receptor for Mac-1 (CD11b/CD18). J Cell Biol. 1990 Dec;111(6 Pt 2):3129–3139. doi: 10.1083/jcb.111.6.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dustin M. L., Springer T. A. Lymphocyte function-associated antigen-1 (LFA-1) interaction with intercellular adhesion molecule-1 (ICAM-1) is one of at least three mechanisms for lymphocyte adhesion to cultured endothelial cells. J Cell Biol. 1988 Jul;107(1):321–331. doi: 10.1083/jcb.107.1.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fischer D. G., Pike M. C., Koren H. S., Snyderman R. Chemotactically responsive and nonresposive forms of a continuous human monocyte cell line. J Immunol. 1980 Jul;125(1):463–465. [PubMed] [Google Scholar]
  19. FitzGerald G. A., Oates J. A., Hawiger J., Maas R. L., Roberts L. J., 2nd, Lawson J. A., Brash A. R. Endogenous biosynthesis of prostacyclin and thromboxane and platelet function during chronic administration of aspirin in man. J Clin Invest. 1983 Mar;71(3):676–688. doi: 10.1172/JCI110814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gamble J. R., Vadas M. A. A new assay for the measurement of the attachment of neutrophils and other cell types to endothelial cells. J Immunol Methods. 1988 May 9;109(2):175–184. doi: 10.1016/0022-1759(88)90240-2. [DOI] [PubMed] [Google Scholar]
  21. Gerrity R. G., Naito H. K. Ultrastructural identification of monocyte-derived foam cells in fatty streak lesions. Artery. 1980;8(3):208–214. [PubMed] [Google Scholar]
  22. Ginsberg M. H., Du X., Plow E. F. Inside-out integrin signalling. Curr Opin Cell Biol. 1992 Oct;4(5):766–771. doi: 10.1016/0955-0674(92)90099-x. [DOI] [PubMed] [Google Scholar]
  23. Goyert S. M., Ferrero E., Rettig W. J., Yenamandra A. K., Obata F., Le Beau M. M. The CD14 monocyte differentiation antigen maps to a region encoding growth factors and receptors. Science. 1988 Jan 29;239(4839):497–500. doi: 10.1126/science.2448876. [DOI] [PubMed] [Google Scholar]
  24. Holme P. A., Orvim U., Hamers M. J., Solum N. O., Brosstad F. R., Barstad R. M., Sakariassen K. S. Shear-induced platelet activation and platelet microparticle formation at blood flow conditions as in arteries with a severe stenosis. Arterioscler Thromb Vasc Biol. 1997 Apr;17(4):646–653. doi: 10.1161/01.atv.17.4.646. [DOI] [PubMed] [Google Scholar]
  25. Issekutz T. B., Issekutz A. C., Movat H. Z. The in vivo quantitation and kinetics of monocyte migration into acute inflammatory tissue. Am J Pathol. 1981 Apr;103(1):47–55. [PMC free article] [PubMed] [Google Scholar]
  26. Iwamoto S., Kawasaki T., Kambayashi J., Ariyoshi H., Monden M. Platelet microparticles: a carrier of platelet-activating factor? Biochem Biophys Res Commun. 1996 Jan 26;218(3):940–944. doi: 10.1006/bbrc.1996.0166. [DOI] [PubMed] [Google Scholar]
  27. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kaplan D. R., Chao F. C., Stiles C. D., Antoniades H. N., Scher C. D. Platelet alpha granules contain a growth factor for fibroblasts. Blood. 1979 Jun;53(6):1043–1052. [PubMed] [Google Scholar]
  29. Lee Y. J., Jy W., Horstman L. L., Janania J., Reyes Y., Kelley R. E., Ahn Y. S. Elevated platelet microparticles in transient ischemic attacks, lacunar infarcts, and multiinfarct dementias. Thromb Res. 1993 Nov 15;72(4):295–304. doi: 10.1016/0049-3848(93)90138-e. [DOI] [PubMed] [Google Scholar]
  30. Leeuwenberg J. F., Smeets E. F., Neefjes J. J., Shaffer M. A., Cinek T., Jeunhomme T. M., Ahern T. J., Buurman W. A. E-selectin and intercellular adhesion molecule-1 are released by activated human endothelial cells in vitro. Immunology. 1992 Dec;77(4):543–549. [PMC free article] [PubMed] [Google Scholar]
  31. Lo S. K., Van Seventer G. A., Levin S. M., Wright S. D. Two leukocyte receptors (CD11a/CD18 and CD11b/CD18) mediate transient adhesion to endothelium by binding to different ligands. J Immunol. 1989 Nov 15;143(10):3325–3329. [PubMed] [Google Scholar]
  32. Mantovani A., Dejana E. Cytokines as communication signals between leukocytes and endothelial cells. Immunol Today. 1989 Nov;10(11):370–375. doi: 10.1016/0167-5699(89)90270-3. [DOI] [PubMed] [Google Scholar]
  33. Molino M., Bainton D. F., Hoxie J. A., Coughlin S. R., Brass L. F. Thrombin receptors on human platelets. Initial localization and subsequent redistribution during platelet activation. J Biol Chem. 1997 Feb 28;272(9):6011–6017. doi: 10.1074/jbc.272.9.6011. [DOI] [PubMed] [Google Scholar]
  34. Nomura S., Komiyama Y., Miyake T., Miyazaki Y., Kido H., Suzuki M., Kagawa H., Yanabu M., Takahashi H., Fukuhara S. Amyloid beta-protein precursor-rich platelet microparticles in thrombotic disease. Thromb Haemost. 1994 Oct;72(4):519–522. [PubMed] [Google Scholar]
  35. Nomura S., Suzuki M., Katsura K., Xie G. L., Miyazaki Y., Miyake T., Kido H., Kagawa H., Fukuhara S. Platelet-derived microparticles may influence the development of atherosclerosis in diabetes mellitus. Atherosclerosis. 1995 Aug;116(2):235–240. doi: 10.1016/0021-9150(95)05551-7. [DOI] [PubMed] [Google Scholar]
  36. Numano F., Kishi Y., Ashikaga T., Kobayashi T., Simokado K., Numano F., Yajima M. Antiplatelet therapy for atherosclerotic disorders. Ann N Y Acad Sci. 1990;598:356–367. doi: 10.1111/j.1749-6632.1990.tb42306.x. [DOI] [PubMed] [Google Scholar]
  37. Olivier A. R., Parker P. J. Identification of multiple PKC isoforms in Swiss 3T3 cells: differential down-regulation by phorbol ester. J Cell Physiol. 1992 Aug;152(2):240–244. doi: 10.1002/jcp.1041520204. [DOI] [PubMed] [Google Scholar]
  38. Pober J. S., Gimbrone M. A., Jr, Lapierre L. A., Mendrick D. L., Fiers W., Rothlein R., Springer T. A. Overlapping patterns of activation of human endothelial cells by interleukin 1, tumor necrosis factor, and immune interferon. J Immunol. 1986 Sep 15;137(6):1893–1896. [PubMed] [Google Scholar]
  39. Reilly I. A., Doran J. B., Smith B., FitzGerald G. A. Increased thromboxane biosynthesis in a human preparation of platelet activation: biochemical and functional consequences of selective inhibition of thromboxane synthase. Circulation. 1986 Jun;73(6):1300–1309. doi: 10.1161/01.cir.73.6.1300. [DOI] [PubMed] [Google Scholar]
  40. Renkonen R. Activation of protein kinases A and C increase lymphocyte penetration through endothelial monolayers. FEBS Lett. 1990 Jul 2;267(1):89–92. doi: 10.1016/0014-5793(90)80295-t. [DOI] [PubMed] [Google Scholar]
  41. Rice G. E., Munro J. M., Bevilacqua M. P. Inducible cell adhesion molecule 110 (INCAM-110) is an endothelial receptor for lymphocytes. A CD11/CD18-independent adhesion mechanism. J Exp Med. 1990 Apr 1;171(4):1369–1374. doi: 10.1084/jem.171.4.1369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ross R. Platelets, platelet-derived growth factor, growth control, and their interactions with the vascular wall. J Cardiovasc Pharmacol. 1985;7 (Suppl 3):S186–S190. doi: 10.1097/00005344-198500073-00022. [DOI] [PubMed] [Google Scholar]
  43. Satta N., Freyssinet J. M., Toti F. The significance of human monocyte thrombomodulin during membrane vesiculation and after stimulation by lipopolysaccharide. Br J Haematol. 1997 Mar;96(3):534–542. doi: 10.1046/j.1365-2141.1997.d01-2059.x. [DOI] [PubMed] [Google Scholar]
  44. Satta N., Toti F., Feugeas O., Bohbot A., Dachary-Prigent J., Eschwège V., Hedman H., Freyssinet J. M. Monocyte vesiculation is a possible mechanism for dissemination of membrane-associated procoagulant activities and adhesion molecules after stimulation by lipopolysaccharide. J Immunol. 1994 Oct 1;153(7):3245–3255. [PubMed] [Google Scholar]
  45. Schwartz L., Bourassa M. G., Lespérance J., Aldridge H. E., Kazim F., Salvatori V. A., Henderson M., Bonan R., David P. R. Aspirin and dipyridamole in the prevention of restenosis after percutaneous transluminal coronary angioplasty. N Engl J Med. 1988 Jun 30;318(26):1714–1719. doi: 10.1056/NEJM198806303182603. [DOI] [PubMed] [Google Scholar]
  46. Serruys P. W., Rutsch W., Heyndrickx G. R., Danchin N., Mast E. G., Wijns W., Rensing B. J., Vos J., Stibbe J. Prevention of restenosis after percutaneous transluminal coronary angioplasty with thromboxane A2-receptor blockade. A randomized, double-blind, placebo-controlled trial. Coronary Artery Restenosis Prevention on Repeated Thromboxane-Antagonism Study (CARPORT). Circulation. 1991 Oct;84(4):1568–1580. doi: 10.1161/01.cir.84.4.1568. [DOI] [PubMed] [Google Scholar]
  47. Shi Y., Kornovski B. S., Savani R., Turley E. A. A rapid, multiwell colorimetric assay for chemotaxis. J Immunol Methods. 1993 Sep 15;164(2):149–154. doi: 10.1016/0022-1759(93)90307-s. [DOI] [PubMed] [Google Scholar]
  48. Sims P. J., Faioni E. M., Wiedmer T., Shattil S. J. Complement proteins C5b-9 cause release of membrane vesicles from the platelet surface that are enriched in the membrane receptor for coagulation factor Va and express prothrombinase activity. J Biol Chem. 1988 Dec 5;263(34):18205–18212. [PubMed] [Google Scholar]
  49. Sims P. J., Wiedmer T., Esmon C. T., Weiss H. J., Shattil S. J. Assembly of the platelet prothrombinase complex is linked to vesiculation of the platelet plasma membrane. Studies in Scott syndrome: an isolated defect in platelet procoagulant activity. J Biol Chem. 1989 Oct 15;264(29):17049–17057. [PubMed] [Google Scholar]
  50. Springer T. A. Adhesion receptors of the immune system. Nature. 1990 Aug 2;346(6283):425–434. doi: 10.1038/346425a0. [DOI] [PubMed] [Google Scholar]
  51. Sundström C., Nilsson K. Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer. 1976 May 15;17(5):565–577. doi: 10.1002/ijc.2910170504. [DOI] [PubMed] [Google Scholar]
  52. Tans G., Rosing J., Thomassen M. C., Heeb M. J., Zwaal R. F., Griffin J. H. Comparison of anticoagulant and procoagulant activities of stimulated platelets and platelet-derived microparticles. Blood. 1991 Jun 15;77(12):2641–2648. [PubMed] [Google Scholar]
  53. Toullec D., Pianetti P., Coste H., Bellevergue P., Grand-Perret T., Ajakane M., Baudet V., Boissin P., Boursier E., Loriolle F. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem. 1991 Aug 25;266(24):15771–15781. [PubMed] [Google Scholar]
  54. Van de Werf F. Clinical trials with glycoprotein IIb/IIIa receptor antagonists in acute coronary syndromes. Thromb Haemost. 1997 Jul;78(1):210–213. [PubMed] [Google Scholar]
  55. Wilentz J. R., Sanborn T. A., Haudenschild C. C., Valeri C. R., Ryan T. J., Faxon D. P. Platelet accumulation in experimental angioplasty: time course and relation to vascular injury. Circulation. 1987 Mar;75(3):636–642. doi: 10.1161/01.cir.75.3.636. [DOI] [PubMed] [Google Scholar]
  56. Zimmerman G. A., McIntyre T. M., Prescott S. M. Adhesion and signaling in vascular cell--cell interactions. J Clin Invest. 1996 Oct 15;98(8):1699–1702. doi: 10.1172/JCI118967. [DOI] [PMC free article] [PubMed] [Google Scholar]

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