Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Feb 2;128(4):661–671. doi: 10.1083/jcb.128.4.661

P-selectin glycoprotein ligand-1 mediates rolling of human neutrophils on P-selectin

PMCID: PMC2199883  PMID: 7532174

Abstract

Neutrophils roll on P-selectin expressed by activated platelets or endothelial cells under the shear stresses in the microcirculation. P- selectin glycoprotein ligand-1 (PSGL-1) is a high affinity ligand for P- selectin on myeloid cells. However, it has not been demonstrated that PSGL-1 contributes to the rolling of neutrophils on P-selectin. We developed two IgG mAbs, PL1 and PL2, that appear to recognize protein- dependent epitopes on human PSGL-1. The mAbs bound to PSGL-1 on all leukocytes as well as on heterologous cells transfected with PSGL-1 cDNA. PL1, but not PL2, blocked binding of 125-I-PSGL-1 to immobilized P-selectin, binding of fluid-phase P-selectin to myeloid and lymphoid leukocytes, adhesion of neutrophils to immobilized P-selectin under static conditions, and rolling of neutrophils on P-selectin-expressing CHO cells under a range of shear stresses. PSGL-1 was localized to microvilli on neutrophils, a topography that may facilitate its adhesive function. These data indicate that (a) PSGL-1 accounts for the high affinity binding sites for P-selectin on leukocytes, and (b) PSGL- 1 must interact with P-selectin in order for neutrophils to roll on P- selectin at physiological shear stresses.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

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

  1. Asada M., Furukawa K., Kantor C., Gahmberg C. G., Kobata A. Structural study of the sugar chains of human leukocyte cell adhesion molecules CD11/CD18. Biochemistry. 1991 Feb 12;30(6):1561–1571. doi: 10.1021/bi00220a017. [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. Baumhueter S., Dybdal N., Kyle C., Lasky L. A. Global vascular expression of murine CD34, a sialomucin-like endothelial ligand for L-selectin. Blood. 1994 Oct 15;84(8):2554–2565. [PubMed] [Google Scholar]
  4. Beesley J. E., Pearson J. D., Hutchings A., Carleton J. S., Gordon J. L. Granulocyte migration through endothelium in culture. J Cell Sci. 1979 Aug;38:237–248. doi: 10.1242/jcs.38.1.237. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Bevilacqua M. P., Nelson R. M. Selectins. J Clin Invest. 1993 Feb;91(2):379–387. doi: 10.1172/JCI116210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Borregaard N., Kjeldsen L., Sengeløv H., Diamond M. S., Springer T. A., Anderson H. C., Kishimoto T. K., Bainton D. F. Changes in subcellular localization and surface expression of L-selectin, alkaline phosphatase, and Mac-1 in human neutrophils during stimulation with inflammatory mediators. J Leukoc Biol. 1994 Jul;56(1):80–87. doi: 10.1002/jlb.56.1.80. [DOI] [PubMed] [Google Scholar]
  8. Briskin M. J., McEvoy L. M., Butcher E. C. MAdCAM-1 has homology to immunoglobulin and mucin-like adhesion receptors and to IgA1. Nature. 1993 Jun 3;363(6428):461–464. doi: 10.1038/363461a0. [DOI] [PubMed] [Google Scholar]
  9. Brustein M., Kraal G., Mebius R. E., Watson S. R. Identification of a soluble form of a ligand for the lymphocyte homing receptor. J Exp Med. 1992 Nov 1;176(5):1415–1419. doi: 10.1084/jem.176.5.1415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Buttrum S. M., Hatton R., Nash G. B. Selectin-mediated rolling of neutrophils on immobilized platelets. Blood. 1993 Aug 15;82(4):1165–1174. [PubMed] [Google Scholar]
  11. Cooper J. A. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Damle N. K., Klussman K., Dietsch M. T., Mohagheghpour N., Aruffo A. GMP-140 (P-selectin/CD62) binds to chronically stimulated but not resting CD4+ T lymphocytes and regulates their production of proinflammatory cytokines. Eur J Immunol. 1992 Jul;22(7):1789–1793. doi: 10.1002/eji.1830220718. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Erlandsen S. L., Hasslen S. R., Nelson R. D. Detection and spatial distribution of the beta 2 integrin (Mac-1) and L-selectin (LECAM-1) adherence receptors on human neutrophils by high-resolution field emission SEM. J Histochem Cytochem. 1993 Mar;41(3):327–333. doi: 10.1177/41.3.7679125. [DOI] [PubMed] [Google Scholar]
  15. Esmon C. T., Esmon N. L., Le Bonniec B. F., Johnson A. E. Protein C activation. Methods Enzymol. 1993;222:359–385. doi: 10.1016/0076-6879(93)22024-a. [DOI] [PubMed] [Google Scholar]
  16. Foxall C., Watson S. R., Dowbenko D., Fennie C., Lasky L. A., Kiso M., Hasegawa A., Asa D., Brandley B. K. The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide. J Cell Biol. 1992 May;117(4):895–902. doi: 10.1083/jcb.117.4.895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Geng J. G., Bevilacqua M. P., Moore K. L., McIntyre T. M., Prescott S. M., Kim J. M., Bliss G. A., Zimmerman G. A., McEver R. P. Rapid neutrophil adhesion to activated endothelium mediated by GMP-140. Nature. 1990 Feb 22;343(6260):757–760. doi: 10.1038/343757a0. [DOI] [PubMed] [Google Scholar]
  18. Hattori R., Hamilton K. K., McEver R. P., Sims P. J. Complement proteins C5b-9 induce secretion of high molecular weight multimers of endothelial von Willebrand factor and translocation of granule membrane protein GMP-140 to the cell surface. J Biol Chem. 1989 May 25;264(15):9053–9060. [PubMed] [Google Scholar]
  19. 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]
  20. Jones D. A., Abbassi O., McIntire L. V., McEver R. P., Smith C. W. P-selectin mediates neutrophil rolling on histamine-stimulated endothelial cells. Biophys J. 1993 Oct;65(4):1560–1569. doi: 10.1016/S0006-3495(93)81195-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kansas G. S., Saunders K. B., Ley K., Zakrzewicz A., Gibson R. M., Furie B. C., Furie B., Tedder T. F. A role for the epidermal growth factor-like domain of P-selectin in ligand recognition and cell adhesion. J Cell Biol. 1994 Feb;124(4):609–618. doi: 10.1083/jcb.124.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuijpers T. W., Hoogerwerf M., van der Laan L. J., Nagel G., van der Schoot C. E., Grunert F., Roos D. CD66 nonspecific cross-reacting antigens are involved in neutrophil adherence to cytokine-activated endothelial cells. J Cell Biol. 1992 Jul;118(2):457–466. doi: 10.1083/jcb.118.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Lee N., Wang W. C., Fukuda M. Granulocytic differentiation of HL-60 cells is associated with increase of poly-N-acetyllactosamine in Asn-linked oligosaccharides attached to human lysosomal membrane glycoproteins. J Biol Chem. 1990 Nov 25;265(33):20476–20487. [PubMed] [Google Scholar]
  26. Lenter M., Levinovitz A., Isenmann S., Vestweber D. Monospecific and common glycoprotein ligands for E- and P-selectin on myeloid cells. J Cell Biol. 1994 Apr;125(2):471–481. doi: 10.1083/jcb.125.2.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Levinovitz A., Mühlhoff J., Isenmann S., Vestweber D. Identification of a glycoprotein ligand for E-selectin on mouse myeloid cells. J Cell Biol. 1993 Apr;121(2):449–459. doi: 10.1083/jcb.121.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ley K., Gaehtgens P. Endothelial, not hemodynamic, differences are responsible for preferential leukocyte rolling in rat mesenteric venules. Circ Res. 1991 Oct;69(4):1034–1041. doi: 10.1161/01.res.69.4.1034. [DOI] [PubMed] [Google Scholar]
  29. Maemura K., Fukuda M. Poly-N-acetyllactosaminyl O-glycans attached to leukosialin. The presence of sialyl Le(x) structures in O-glycans. J Biol Chem. 1992 Dec 5;267(34):24379–24386. [PubMed] [Google Scholar]
  30. Marlin S. D., Springer T. A. Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen 1 (LFA-1). Cell. 1987 Dec 4;51(5):813–819. doi: 10.1016/0092-8674(87)90104-8. [DOI] [PubMed] [Google Scholar]
  31. McEver R. P., Martin M. N. A monoclonal antibody to a membrane glycoprotein binds only to activated platelets. J Biol Chem. 1984 Aug 10;259(15):9799–9804. [PubMed] [Google Scholar]
  32. McEver R. P. Selectins. Curr Opin Immunol. 1994 Feb;6(1):75–84. doi: 10.1016/0952-7915(94)90037-x. [DOI] [PubMed] [Google Scholar]
  33. Moore K. L., Stults N. L., Diaz S., Smith D. F., Cummings R. D., Varki A., McEver R. P. Identification of a specific glycoprotein ligand for P-selectin (CD62) on myeloid cells. J Cell Biol. 1992 Jul;118(2):445–456. doi: 10.1083/jcb.118.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Moore K. L., Thompson L. F. P-selectin (CD62) binds to subpopulations of human memory T lymphocytes and natural killer cells. Biochem Biophys Res Commun. 1992 Jul 15;186(1):173–181. doi: 10.1016/s0006-291x(05)80790-9. [DOI] [PubMed] [Google Scholar]
  35. Moore K. L., Varki A., McEver R. P. GMP-140 binds to a glycoprotein receptor on human neutrophils: evidence for a lectin-like interaction. J Cell Biol. 1991 Feb;112(3):491–499. doi: 10.1083/jcb.112.3.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Norgard-Sumnicht K. E., Varki N. M., Varki A. Calcium-dependent heparin-like ligands for L-selectin in nonlymphoid endothelial cells. Science. 1993 Jul 23;261(5120):480–483. doi: 10.1126/science.7687382. [DOI] [PubMed] [Google Scholar]
  37. Norgard K. E., Moore K. L., Diaz S., Stults N. L., Ushiyama S., McEver R. P., Cummings R. D., Varki A. Characterization of a specific ligand for P-selectin on myeloid cells. A minor glycoprotein with sialylated O-linked oligosaccharides. J Biol Chem. 1993 Jun 15;268(17):12764–12774. [PubMed] [Google Scholar]
  38. 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]
  39. Postigo A. A., Marazuela M., Sánchez-Madrid F., de Landázuri M. O. B lymphocyte binding to E- and P-selectins is mediated through the de novo expression of carbohydrates on in vitro and in vivo activated human B cells. J Clin Invest. 1994 Oct;94(4):1585–1596. doi: 10.1172/JCI117500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sako D., Chang X. J., Barone K. M., Vachino G., White H. M., Shaw G., Veldman G. M., Bean K. M., Ahern T. J., Furie B. Expression cloning of a functional glycoprotein ligand for P-selectin. Cell. 1993 Dec 17;75(6):1179–1186. doi: 10.1016/0092-8674(93)90327-m. [DOI] [PubMed] [Google Scholar]
  41. Smith C. W., Hollers J. C. Motility and adhesiveness in human neutrophils. Redistribution of chemotactic factor-induced adhesion sites. J Clin Invest. 1980 Apr;65(4):804–812. doi: 10.1172/JCI109731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Steininger C. N., Eddy C. A., Leimgruber R. M., Mellors A., Welply J. K. The glycoprotease of Pasteurella haemolytica A1 eliminates binding of myeloid cells to P-selectin but not to E-selectin. Biochem Biophys Res Commun. 1992 Oct 30;188(2):760–766. doi: 10.1016/0006-291x(92)91121-6. [DOI] [PubMed] [Google Scholar]
  43. Sutherland D. R., Abdullah K. M., Cyopick P., Mellors A. Cleavage of the cell-surface O-sialoglycoproteins CD34, CD43, CD44, and CD45 by a novel glycoprotease from Pasteurella haemolytica. J Immunol. 1992 Mar 1;148(5):1458–1464. [PubMed] [Google Scholar]
  44. Uetsuki T., Naito A., Nagata S., Kaziro Y. Isolation and characterization of the human chromosomal gene for polypeptide chain elongation factor-1 alpha. J Biol Chem. 1989 Apr 5;264(10):5791–5798. [PubMed] [Google Scholar]
  45. Ushiyama S., Laue T. M., Moore K. L., Erickson H. P., McEver R. P. Structural and functional characterization of monomeric soluble P-selectin and comparison with membrane P-selectin. J Biol Chem. 1993 Jul 15;268(20):15229–15237. [PubMed] [Google Scholar]
  46. Varki A. Selectin ligands. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7390–7397. doi: 10.1073/pnas.91.16.7390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Varki A. Selectins and other mammalian sialic acid-binding lectins. Curr Opin Cell Biol. 1992 Apr;4(2):257–266. doi: 10.1016/0955-0674(92)90041-a. [DOI] [PubMed] [Google Scholar]
  48. Walcheck B., Watts G., Jutila M. A. Bovine gamma/delta T cells bind E-selectin via a novel glycoprotein receptor: first characterization of a lymphocyte/E-selectin interaction in an animal model. J Exp Med. 1993 Sep 1;178(3):853–863. doi: 10.1084/jem.178.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wright S. D., Rao P. E., Van Voorhis W. C., Craigmyle L. S., Iida K., Talle M. A., Westberg E. F., Goldstein G., Silverstein S. C. Identification of the C3bi receptor of human monocytes and macrophages by using monoclonal antibodies. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5699–5703. doi: 10.1073/pnas.80.18.5699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Zhou Q., Moore K. L., Smith D. F., Varki A., McEver R. P., Cummings R. D. The selectin GMP-140 binds to sialylated, fucosylated lactosaminoglycans on both myeloid and nonmyeloid cells. J Cell Biol. 1991 Oct;115(2):557–564. doi: 10.1083/jcb.115.2.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Zimmerman G. A., McIntyre T. M., Prescott S. M. Thrombin stimulates the adherence of neutrophils to human endothelial cells in vitro. J Clin Invest. 1985 Dec;76(6):2235–2246. doi: 10.1172/JCI112232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. de Bruijne-Admiraal L. G., Modderman P. W., Von dem Borne A. E., Sonnenberg A. P-selectin mediates Ca(2+)-dependent adhesion of activated platelets to many different types of leukocytes: detection by flow cytometry. Blood. 1992 Jul 1;80(1):134–142. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES