Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Sep 1;98(5):1081–1087. doi: 10.1172/JCI118888

Neutrophil-neutrophil interactions under hydrodynamic shear stress involve L-selectin and PSGL-1. A mechanism that amplifies initial leukocyte accumulation of P-selectin in vitro.

B Walcheck 1, K L Moore 1, R P McEver 1, T K Kishimoto 1
PMCID: PMC507527  PMID: 8787668

Abstract

Leukocytes attach to and roll on inflamed endothelium and on leukocyte monolayers that form on the endothelial cells. Leukocyte-leukocyte interactions occurring under hydrodynamic shear stress are mediated by binding of L-selectin to unknown sialomucin-like glycoproteins. We show that purified neutrophil PSGL-1, a sialomucin glycoprotein that serves as a ligand for both P- and E-selectin, can also support the attachment and rolling of free flowing neutrophils in vitro. Neutrophil rolling on PSGL-1 was abolished by the anti-L-selectin mAb DREG200 and by the anti-PSGL-1 mAb PL1, indicating that L-selectin can interact directly with PSGL-1. Neutrophil rolling on neutrophil monolayers was also blocked by PL1 (60 +/- 9% SEM inhibition); however, DREG200 blocked more efficiently (93 +/- 7% SEM inhibition), suggesting that other L-selectin ligands may exist on the neutrophil surface. These studies demonstrate that PSGL-1 on the neutrophil surface is a major functional ligand for L-selectin. The avidity of this L-selectin-dependent adhesion event was sufficient to allow individual neutrophils rolling on P-selectin to capture free flowing neutrophils, which progressed to form linear strings and discrete foci of rolling neutrophils. Neutrophil accumulation on P-selectin accelerated with time as a result of neutrophil-assisted capture of free flowing neutrophils. When neutrophil-neutrophil interactions were blocked by DREG200, neutrophils accumulated on P-selectin in a random pattern and at a uniform rate. Thus, leukocyte-assisted capture of flowing leukocytes may play an important role in amplifying the rate of initial leukocyte recruitment at sites of inflammation.

Full Text

The Full Text of this article is available as a PDF (554.6 KB).

Selected References

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

  1. Alon R., Hammer D. A., Springer T. A. Lifetime of the P-selectin-carbohydrate bond and its response to tensile force in hydrodynamic flow. Nature. 1995 Apr 6;374(6522):539–542. doi: 10.1038/374539a0. [DOI] [PubMed] [Google Scholar]
  2. Asa D., Raycroft L., Ma L., Aeed P. A., Kaytes P. S., Elhammer A. P., Geng J. G. The P-selectin glycoprotein ligand functions as a common human leukocyte ligand for P- and E-selectins. J Biol Chem. 1995 May 12;270(19):11662–11670. doi: 10.1074/jbc.270.19.11662. [DOI] [PubMed] [Google Scholar]
  3. Bargatze R. F., Kurk S., Butcher E. C., Jutila M. A. Neutrophils roll on adherent neutrophils bound to cytokine-induced endothelial cells via L-selectin on the rolling cells. J Exp Med. 1994 Nov 1;180(5):1785–1792. doi: 10.1084/jem.180.5.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Bennett T. A., Schammel C. M., Lynam E. B., Guyer D. A., Mellors A., Edwards B., Rogelj S., Sklar L. A. Evidence for a third component in neutrophil aggregation: potential roles of O-linked glycoproteins as L-selectin counter-structures. J Leukoc Biol. 1995 Nov;58(5):510–518. doi: 10.1002/jlb.58.5.510. [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. Bosse R., Vestweber D. Only simultaneous blocking of the L- and P-selectin completely inhibits neutrophil migration into mouse peritoneum. Eur J Immunol. 1994 Dec;24(12):3019–3024. doi: 10.1002/eji.1830241215. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Finger E. B., Puri K. D., Alon R., Lawrence M. B., von Andrian U. H., Springer T. A. Adhesion through L-selectin requires a threshold hydrodynamic shear. Nature. 1996 Jan 18;379(6562):266–269. doi: 10.1038/379266a0. [DOI] [PubMed] [Google Scholar]
  11. Gibbs J. B. Ras C-terminal processing enzymes--new drug targets? Cell. 1991 Apr 5;65(1):1–4. doi: 10.1016/0092-8674(91)90352-y. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Jutila M. A., Bargatze R. F., Kurk S., Warnock R. A., Ehsani N., Watson S. R., Walcheck B. Cell surface P- and E-selectin support shear-dependent rolling of bovine gamma/delta T cells. J Immunol. 1994 Nov 1;153(9):3917–3928. [PubMed] [Google Scholar]
  14. Jutila M. A., Kurk S. Analysis of bovine gamma delta T cell interactions with E-, P-, and L-selectin. Characterization of lymphocyte on lymphocyte rolling and the effects of O-glycoprotease. J Immunol. 1996 Jan 1;156(1):289–296. [PubMed] [Google Scholar]
  15. 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]
  16. Kishimoto T. K., Jutila M. A., Butcher E. C. Identification of a human peripheral lymph node homing receptor: a rapidly down-regulated adhesion molecule. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2244–2248. doi: 10.1073/pnas.87.6.2244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kishimoto T. K., Warnock R. A., Jutila M. A., Butcher E. C., Lane C., Anderson D. C., Smith C. W. Antibodies against human neutrophil LECAM-1 (LAM-1/Leu-8/DREG-56 antigen) and endothelial cell ELAM-1 inhibit a common CD18-independent adhesion pathway in vitro. Blood. 1991 Aug 1;78(3):805–811. [PubMed] [Google Scholar]
  18. 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]
  19. Lawrence M. B., Bainton D. F., Springer T. A. Neutrophil tethering to and rolling on E-selectin are separable by requirement for L-selectin. Immunity. 1994 May;1(2):137–145. doi: 10.1016/1074-7613(94)90107-4. [DOI] [PubMed] [Google Scholar]
  20. Lawrence M. B., Berg E. L., Butcher E. C., Springer T. A. Rolling of lymphocytes and neutrophils on peripheral node addressin and subsequent arrest on ICAM-1 in shear flow. Eur J Immunol. 1995 Apr;25(4):1025–1031. doi: 10.1002/eji.1830250425. [DOI] [PubMed] [Google Scholar]
  21. Lawrence M. B., Springer T. A. Neutrophils roll on E-selectin. J Immunol. 1993 Dec 1;151(11):6338–6346. [PubMed] [Google Scholar]
  22. 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]
  23. Li F., Erickson H. P., James J. A., Moore K. L., Cummings R. D., McEver R. P. Visualization of P-selectin glycoprotein ligand-1 as a highly extended molecule and mapping of protein epitopes for monoclonal antibodies. J Biol Chem. 1996 Mar 15;271(11):6342–6348. doi: 10.1074/jbc.271.11.6342. [DOI] [PubMed] [Google Scholar]
  24. Li F., Wilkins P. P., Crawley S., Weinstein J., Cummings R. D., McEver R. P. Post-translational modifications of recombinant P-selectin glycoprotein ligand-1 required for binding to P- and E-selectin. J Biol Chem. 1996 Feb 9;271(6):3255–3264. [PubMed] [Google Scholar]
  25. Ma X. L., Weyrich A. S., Lefer D. J., Buerke M., Albertine K. H., Kishimoto T. K., Lefer A. M. Monoclonal antibody to L-selectin attenuates neutrophil accumulation and protects ischemic reperfused cat myocardium. Circulation. 1993 Aug;88(2):649–658. doi: 10.1161/01.cir.88.2.649. [DOI] [PubMed] [Google Scholar]
  26. McEver R. P., Moore K. L., Cummings R. D. Leukocyte trafficking mediated by selectin-carbohydrate interactions. J Biol Chem. 1995 May 12;270(19):11025–11028. doi: 10.1074/jbc.270.19.11025. [DOI] [PubMed] [Google Scholar]
  27. Moore K. L., Eaton S. F., Lyons D. E., Lichenstein H. S., Cummings R. D., McEver R. P. The P-selectin glycoprotein ligand from human neutrophils displays sialylated, fucosylated, O-linked poly-N-acetyllactosamine. J Biol Chem. 1994 Sep 16;269(37):23318–23327. [PubMed] [Google Scholar]
  28. Moore K. L., Patel K. D., Bruehl R. E., Li F., Johnson D. A., Lichenstein H. S., Cummings R. D., Bainton D. F., McEver R. P. P-selectin glycoprotein ligand-1 mediates rolling of human neutrophils on P-selectin. J Cell Biol. 1995 Feb;128(4):661–671. doi: 10.1083/jcb.128.4.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Mulligan M. S., Miyasaka M., Tamatani T., Jones M. L., Ward P. A. Requirements for L-selectin in neutrophil-mediated lung injury in rats. J Immunol. 1994 Jan 15;152(2):832–840. [PubMed] [Google Scholar]
  32. Mulligan M. S., Till G. O., Smith C. W., Anderson D. C., Miyasaka M., Tamatani T., Todd R. F., 3rd, Issekutz T. B., Ward P. A. Role of leukocyte adhesion molecules in lung and dermal vascular injury after thermal trauma of skin. Am J Pathol. 1994 May;144(5):1008–1015. [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Norman K. E., Moore K. L., McEver R. P., Ley K. Leukocyte rolling in vivo is mediated by P-selectin glycoprotein ligand-1. Blood. 1995 Dec 15;86(12):4417–4421. [PubMed] [Google Scholar]
  35. Patel K. D., Moore K. L., Nollert M. U., McEver R. P. Neutrophils use both shared and distinct mechanisms to adhere to selectins under static and flow conditions. J Clin Invest. 1995 Oct;96(4):1887–1896. doi: 10.1172/JCI118234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Pizcueta P., Luscinskas F. W. Monoclonal antibody blockade of L-selectin inhibits mononuclear leukocyte recruitment to inflammatory sites in vivo. Am J Pathol. 1994 Aug;145(2):461–469. [PMC free article] [PubMed] [Google Scholar]
  38. Pouyani T., Seed B. PSGL-1 recognition of P-selectin is controlled by a tyrosine sulfation consensus at the PSGL-1 amino terminus. Cell. 1995 Oct 20;83(2):333–343. doi: 10.1016/0092-8674(95)90174-4. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Sako D., Comess K. M., Barone K. M., Camphausen R. T., Cumming D. A., Shaw G. D. A sulfated peptide segment at the amino terminus of PSGL-1 is critical for P-selectin binding. Cell. 1995 Oct 20;83(2):323–331. doi: 10.1016/0092-8674(95)90173-6. [DOI] [PubMed] [Google Scholar]
  41. Seekamp A., Till G. O., Mulligan M. S., Paulson J. C., Anderson D. C., Miyasaka M., Ward P. A. Role of selectins in local and remote tissue injury following ischemia and reperfusion. Am J Pathol. 1994 Mar;144(3):592–598. [PMC free article] [PubMed] [Google Scholar]
  42. Simon S. I., Chambers J. D., Butcher E., Sklar L. A. Neutrophil aggregation is beta 2-integrin- and L-selectin-dependent in blood and isolated cells. J Immunol. 1992 Oct 15;149(8):2765–2771. [PubMed] [Google Scholar]
  43. Smith C. W., Rothlein R., Hughes B. J., Mariscalco M. M., Rudloff H. E., Schmalstieg F. C., Anderson D. C. Recognition of an endothelial determinant for CD 18-dependent human neutrophil adherence and transendothelial migration. J Clin Invest. 1988 Nov;82(5):1746–1756. doi: 10.1172/JCI113788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tedder T. F., Steeber D. A., Pizcueta P. L-selectin-deficient mice have impaired leukocyte recruitment into inflammatory sites. J Exp Med. 1995 Jun 1;181(6):2259–2264. doi: 10.1084/jem.181.6.2259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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]
  46. Walcheck B., Kahn J., Fisher J. M., Wang B. B., Fisk R. S., Payan D. G., Feehan C., Betageri R., Darlak K., Spatola A. F. Neutrophil rolling altered by inhibition of L-selectin shedding in vitro. Nature. 1996 Apr 25;380(6576):720–723. doi: 10.1038/380720a0. [DOI] [PubMed] [Google Scholar]
  47. Wilkins P. P., Moore K. L., McEver R. P., Cummings R. D. Tyrosine sulfation of P-selectin glycoprotein ligand-1 is required for high affinity binding to P-selectin. J Biol Chem. 1995 Sep 29;270(39):22677–22680. doi: 10.1074/jbc.270.39.22677. [DOI] [PubMed] [Google Scholar]
  48. von Andrian U. H., Hasslen S. R., Nelson R. D., Erlandsen S. L., Butcher E. C. A central role for microvillous receptor presentation in leukocyte adhesion under flow. Cell. 1995 Sep 22;82(6):989–999. doi: 10.1016/0092-8674(95)90278-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES