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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 Jan 1;101(1):212–222. doi: 10.1172/JCI269

Engagement of human PECAM-1 (CD31) on human endothelial cells increases intracellular calcium ion concentration and stimulates prostacyclin release.

I Gurubhagavatula 1, Y Amrani 1, D Pratico 1, F L Ruberg 1, S M Albelda 1, R A Panettieri Jr 1
PMCID: PMC508558  PMID: 9421484

Abstract

Platelet-endothelial cell adhesion molecule-1 (PECAM-1) is a member of the immunoglobulin superfamily that plays a role in a number of endothelial cell (EC) functions including migration, angiogenesis, and transmigration of leukocytes across endothelium. We postulated that one way PECAM-1 might exert its effects was by regulating intracellular EC levels of calcium. Using single-cell fluorometry, we found that engagement of PECAM-1 by mAbs induced a slow but sustained increase in intracellular calcium, both in EC and in an adherent PECAM-1-transfected cell line that models endothelium. Generation of this signal was specific for certain anti-PECAM-1 antibodies, required the presence of the cytoplasmic domain, depended on extracellular calcium and on tyrosine phosphorylation, but did not require cross-linking; in fact, calcium increases were stimulated by certain Fab fragments. Activation of EC by PECAM-1 also caused a time-dependent increase in prostacyclin release. Given the importance of intracellular calcium and prostacyclin release as signaling molecules, engagement of PECAM-1 during cell-cell interactions may alter a number of EC functions including secretion of vasoactive mediators.

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Selected References

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  1. Albelda S. M., Daise M., Levine E. M., Buck C. A. Identification and characterization of cell-substratum adhesion receptors on cultured human endothelial cells. J Clin Invest. 1989 Jun;83(6):1992–2002. doi: 10.1172/JCI114109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albelda S. M., Muller W. A., Buck C. A., Newman P. J. Molecular and cellular properties of PECAM-1 (endoCAM/CD31): a novel vascular cell-cell adhesion molecule. J Cell Biol. 1991 Sep;114(5):1059–1068. doi: 10.1083/jcb.114.5.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett J., Weeds A. Calcium and the cytoskeleton. Br Med Bull. 1986 Oct;42(4):385–390. doi: 10.1093/oxfordjournals.bmb.a072156. [DOI] [PubMed] [Google Scholar]
  4. Berman M. E., Xie Y., Muller W. A. Roles of platelet/endothelial cell adhesion molecule-1 (PECAM-1, CD31) in natural killer cell transendothelial migration and beta 2 integrin activation. J Immunol. 1996 Feb 15;156(4):1515–1524. [PubMed] [Google Scholar]
  5. Birch K. A., Ewenstein B. M., Golan D. E., Pober J. S. Prolonged peak elevations in cytoplasmic free calcium ions, derived from intracellular stores, correlate with the extent of thrombin-stimulated exocytosis in single human umbilical vein endothelial cells. J Cell Physiol. 1994 Sep;160(3):545–554. doi: 10.1002/jcp.1041600318. [DOI] [PubMed] [Google Scholar]
  6. Buchan K. W., Martin W. Bradykinin induces elevations of cytosolic calcium through mobilisation of intracellular and extracellular pools in bovine aortic endothelial cells. Br J Pharmacol. 1991 Jan;102(1):35–40. doi: 10.1111/j.1476-5381.1991.tb12128.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DeLisser H. M., Chilkotowsky J., Yan H. C., Daise M. L., Buck C. A., Albelda S. M. Deletions in the cytoplasmic domain of platelet-endothelial cell adhesion molecule-1 (PECAM-1, CD31) result in changes in ligand binding properties. J Cell Biol. 1994 Jan;124(1-2):195–203. doi: 10.1083/jcb.124.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DeLisser H. M., Christofidou-Solomidou M., Strieter R. M., Burdick M. D., Robinson C. S., Wexler R. S., Kerr J. S., Garlanda C., Merwin J. R., Madri J. A. Involvement of endothelial PECAM-1/CD31 in angiogenesis. Am J Pathol. 1997 Sep;151(3):671–677. [PMC free article] [PubMed] [Google Scholar]
  9. DeLisser H. M., Newman P. J., Albelda S. M. Molecular and functional aspects of PECAM-1/CD31. Immunol Today. 1994 Oct;15(10):490–495. doi: 10.1016/0167-5699(94)90195-3. [DOI] [PubMed] [Google Scholar]
  10. Deaglio S., Dianzani U., Horenstein A. L., Fernández J. E., van Kooten C., Bragardo M., Funaro A., Garbarino G., Di Virgilio F., Banchereau J. Human CD38 ligand. A 120-KDA protein predominantly expressed on endothelial cells. J Immunol. 1996 Jan 15;156(2):727–734. [PubMed] [Google Scholar]
  11. Dolor R. J., Hurwitz L. M., Mirza Z., Strauss H. C., Whorton A. R. Regulation of extracellular calcium entry in endothelial cells: role of intracellular calcium pool. Am J Physiol. 1992 Jan;262(1 Pt 1):C171–C181. doi: 10.1152/ajpcell.1992.262.1.C171. [DOI] [PubMed] [Google Scholar]
  12. Ferrero E., Ferrero M. E., Pardi R., Zocchi M. R. The platelet endothelial cell adhesion molecule-1 (PECAM1) contributes to endothelial barrier function. FEBS Lett. 1995 Nov 6;374(3):323–326. doi: 10.1016/0014-5793(95)01110-z. [DOI] [PubMed] [Google Scholar]
  13. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  14. Hallam T. J., Jacob R., Merritt J. E. Evidence that agonists stimulate bivalent-cation influx into human endothelial cells. Biochem J. 1988 Oct 1;255(1):179–184. doi: 10.1042/bj2550179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hattori R., Hamilton K. K., Fugate R. D., McEver R. P., Sims P. J. Stimulated secretion of endothelial von Willebrand factor is accompanied by rapid redistribution to the cell surface of the intracellular granule membrane protein GMP-140. J Biol Chem. 1989 May 15;264(14):7768–7771. [PubMed] [Google Scholar]
  16. He P., Pagakis S. N., Curry F. E. Measurement of cytoplasmic calcium in single microvessels with increased permeability. Am J Physiol. 1990 May;258(5 Pt 2):H1366–H1374. doi: 10.1152/ajpheart.1990.258.5.H1366. [DOI] [PubMed] [Google Scholar]
  17. Himmel H. M., Whorton A. R., Strauss H. C. Intracellular calcium, currents, and stimulus-response coupling in endothelial cells. Hypertension. 1993 Jan;21(1):112–127. doi: 10.1161/01.hyp.21.1.112. [DOI] [PubMed] [Google Scholar]
  18. Huang A. J., Manning J. E., Bandak T. M., Ratau M. C., Hanser K. R., Silverstein S. C. Endothelial cell cytosolic free calcium regulates neutrophil migration across monolayers of endothelial cells. J Cell Biol. 1993 Mar;120(6):1371–1380. doi: 10.1083/jcb.120.6.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Itoh K., Kawamura H., Asou H. A novel monoclonal antibody against carbohydrates of L1 cell adhesion molecule causes an influx of calcium in cultured cortical neurons. Brain Res. 1992 May 15;580(1-2):233–240. doi: 10.1016/0006-8993(92)90949-a. [DOI] [PubMed] [Google Scholar]
  20. Jackson D. E., Ward C. M., Wang R., Newman P. J. The protein-tyrosine phosphatase SHP-2 binds platelet/endothelial cell adhesion molecule-1 (PECAM-1) and forms a distinct signaling complex during platelet aggregation. Evidence for a mechanistic link between PECAM-1- and integrin-mediated cellular signaling. J Biol Chem. 1997 Mar 14;272(11):6986–6993. doi: 10.1074/jbc.272.11.6986. [DOI] [PubMed] [Google Scholar]
  21. Juan M., Viñas O., Pino-Otín M. R., Places L., Martínez-Cáceres E., Barceló J. J., Miralles A., Vilella R., de la Fuente M. A., Vives J. CD50 (intercellular adhesion molecule 3) stimulation induces calcium mobilization and tyrosine phosphorylation through p59fyn and p56lck in Jurkat T cell line. J Exp Med. 1994 Jun 1;179(6):1747–1756. doi: 10.1084/jem.179.6.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lu T. T., Yan L. G., Madri J. A. Integrin engagement mediates tyrosine dephosphorylation on platelet-endothelial cell adhesion molecule 1. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11808–11813. doi: 10.1073/pnas.93.21.11808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lum H., Del Vecchio P. J., Schneider A. S., Goligorsky M. S., Malik A. B. Calcium dependence of the thrombin-induced increase in endothelial albumin permeability. J Appl Physiol (1985) 1989 Mar;66(3):1471–1476. doi: 10.1152/jappl.1989.66.3.1471. [DOI] [PubMed] [Google Scholar]
  24. Lum H., Malik A. B. Regulation of vascular endothelial barrier function. Am J Physiol. 1994 Sep;267(3 Pt 1):L223–L241. doi: 10.1152/ajplung.1994.267.3.L223. [DOI] [PubMed] [Google Scholar]
  25. Muller W. A., Weigl S. A., Deng X., Phillips D. M. PECAM-1 is required for transendothelial migration of leukocytes. J Exp Med. 1993 Aug 1;178(2):449–460. doi: 10.1084/jem.178.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Muller W. A., Weigl S. A. Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay. J Exp Med. 1992 Sep 1;176(3):819–828. doi: 10.1084/jem.176.3.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Newman P. J. The biology of PECAM-1. J Clin Invest. 1997 Jan 1;99(1):3–8. doi: 10.1172/JCI119129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Panettieri R. A., Jr, Hall I. P., Maki C. S., Murray R. K. alpha-Thrombin increases cytosolic calcium and induces human airway smooth muscle cell proliferation. Am J Respir Cell Mol Biol. 1995 Aug;13(2):205–216. doi: 10.1165/ajrcmb.13.2.7626288. [DOI] [PubMed] [Google Scholar]
  29. Poggi A., Panzeri M. C., Moretta L., Zocchi M. R. CD31-triggered rearrangement of the actin cytoskeleton in human natural killer cells. Eur J Immunol. 1996 Apr;26(4):817–824. doi: 10.1002/eji.1830260414. [DOI] [PubMed] [Google Scholar]
  30. Pollock W. K., Wreggett K. A., Irvine R. F. Inositol phosphate production and Ca2+ mobilization in human umbilical-vein endothelial cells stimulated by thrombin and histamine. Biochem J. 1988 Dec 1;256(2):371–376. doi: 10.1042/bj2560371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pratico D., Lawson J. A., FitzGerald G. A. Cyclooxygenase-dependent formation of the isoprostane, 8-epi prostaglandin F2 alpha. J Biol Chem. 1995 Apr 28;270(17):9800–9808. doi: 10.1074/jbc.270.17.9800. [DOI] [PubMed] [Google Scholar]
  32. Ryan U. S., Avdonin P. V., Posin E. Y., Popov E. G., Danilov S. M., Tkachuk V. A. Influence of vasoactive agents on cytoplasmic free calcium in vascular endothelial cells. J Appl Physiol (1985) 1988 Nov;65(5):2221–2227. doi: 10.1152/jappl.1988.65.5.2221. [DOI] [PubMed] [Google Scholar]
  33. Schimmenti L. A., Yan H. C., Madri J. A., Albelda S. M. Platelet endothelial cell adhesion molecule, PECAM-1, modulates cell migration. J Cell Physiol. 1992 Nov;153(2):417–428. doi: 10.1002/jcp.1041530222. [DOI] [PubMed] [Google Scholar]
  34. Smythe W. R., Hwang H. C., Amin K. M., Eck S. L., Davidson B. L., Wilson J. M., Kaiser L. R., Albelda S. M. Use of recombinant adenovirus to transfer the herpes simplex virus thymidine kinase (HSVtk) gene to thoracic neoplasms: an effective in vitro drug sensitization system. Cancer Res. 1994 Apr 15;54(8):2055–2059. [PubMed] [Google Scholar]
  35. Spinozzi F., Agea E., Bistoni O., Belia S., Travetti A., Gerli R., Muscat C., Bertotto A. Intracellular calcium levels are differentially regulated in T lymphocytes triggered by anti-CD2 and anti-CD3 monoclonal antibodies. Cell Signal. 1995 Mar;7(3):287–293. doi: 10.1016/0898-6568(94)00079-q. [DOI] [PubMed] [Google Scholar]
  36. Sun J., Williams J., Yan H. C., Amin K. M., Albelda S. M., DeLisser H. M. Platelet endothelial cell adhesion molecule-1 (PECAM-1) homophilic adhesion is mediated by immunoglobulin-like domains 1 and 2 and depends on the cytoplasmic domain and the level of surface expression. J Biol Chem. 1996 Aug 2;271(31):18561–18570. doi: 10.1074/jbc.271.31.18561. [DOI] [PubMed] [Google Scholar]
  37. Sun Q. H., DeLisser H. M., Zukowski M. M., Paddock C., Albelda S. M., Newman P. J. Individually distinct Ig homology domains in PECAM-1 regulate homophilic binding and modulate receptor affinity. J Biol Chem. 1996 May 10;271(19):11090–11098. doi: 10.1074/jbc.271.19.11090. [DOI] [PubMed] [Google Scholar]
  38. Vaporciyan A. A., DeLisser H. M., Yan H. C., Mendiguren I. I., Thom S. R., Jones M. L., Ward P. A., Albelda S. M. Involvement of platelet-endothelial cell adhesion molecule-1 in neutrophil recruitment in vivo. Science. 1993 Dec 3;262(5139):1580–1582. doi: 10.1126/science.8248808. [DOI] [PubMed] [Google Scholar]
  39. Wysolmerski R. B., Lagunoff D. Involvement of myosin light-chain kinase in endothelial cell retraction. Proc Natl Acad Sci U S A. 1990 Jan;87(1):16–20. doi: 10.1073/pnas.87.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yan H. C., Pilewski J. M., Zhang Q., DeLisser H. M., Romer L., Albelda S. M. Localization of multiple functional domains on human PECAM-1 (CD31) by monoclonal antibody epitope mapping. Cell Adhes Commun. 1995 Feb;3(1):45–66. doi: 10.3109/15419069509081277. [DOI] [PubMed] [Google Scholar]

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