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. 1996 Oct 1;184(4):1213–1223. doi: 10.1084/jem.184.4.1213

Human leukocyte elastase is an endogenous ligand for the integrin CR3 (CD11b/CD18, Mac-1, alpha M beta 2) and modulates polymorphonuclear leukocyte adhesion

PMCID: PMC2192826  PMID: 8879192

Abstract

Integrin CR3 (CD11b/CD18, Mac-1, alpha M beta 2) mediates the transient adhesion of polymorphonuclear leukocytes (PMN) to surfaces coated with fibrinogen, C3bi, ICAM-1, and other ligands. Recent studies (Cai, T.- Q., and S.D. Wright 1995. J. Biol. Chem. 270:14358) suggest that adhesion may be favored by stimulus-dependent changes in the kinetics of ligand binding by CR3. Cell detachment, on the other hand, must occur by a different mechanism because binding kinetics cannot affect cell adhesion after binding of ligand has occurred. We have sought a mechanism that would reverse binding of ligand to CR3 and report here that lysates of PMN contain an endogenous ligand that binds CR3 and competes the binding of C3bi. Purification and sequence analysis identified the structurally homologous azurophilic granule proteins, elastase, protease 3, and azurocidin as candidates. Studies with purified elastase and azurocidin showed that each bound specifically to purified, immobilized CR3. Elastase may play a role in modulating integrin-mediated cell adhesion because it is expressed at the cell surface, and the expression level is inversely proportional to cell adhesivity. Furthermore, a monoclonal antibody against elastase prevented detachment of PMN from fibrinogen-coated surfaces and blocked chemotaxis, confirming a role for this protein in regulating integrin- mediated adhesion. These studies suggest a model for release of integrin-mediated cell adhesion in which endogenous ligands such as elastase may release adhesion by "'eluting" substrate-bound ligand. A role for the proteolytic activity of elastase appears likely but is not demonstrated in this study.

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Selected References

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  1. Almeida R. P., Melchior M., Campanelli D., Nathan C., Gabay J. E. Complementary DNA sequence of human neutrophil azurocidin, an antibiotic with extensive homology to serine proteases. Biochem Biophys Res Commun. 1991 Jun 14;177(2):688–695. doi: 10.1016/0006-291x(91)91843-2. [DOI] [PubMed] [Google Scholar]
  2. Altieri D. C., Edgington T. S. The saturable high affinity association of factor X to ADP-stimulated monocytes defines a novel function of the Mac-1 receptor. J Biol Chem. 1988 May 25;263(15):7007–7015. [PubMed] [Google Scholar]
  3. Altieri D. C., Morrissey J. H., Edgington T. S. Adhesive receptor Mac-1 coordinates the activation of factor X on stimulated cells of monocytic and myeloid differentiation: an alternative initiation of the coagulation protease cascade. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7462–7466. doi: 10.1073/pnas.85.20.7462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ashmun R. A., Peiper S. C., Rebentisch M. B., Look A. T. Expression of the human monocyte membrane antigen gp55 by murine fibroblasts after DNA-mediated gene transfer. Blood. 1987 Mar;69(3):886–892. [PubMed] [Google Scholar]
  5. Berger M. Inflammation in the lung in cystic fibrosis. A vicious cycle that does more harm than good? Clin Rev Allergy. 1991 Spring-Summer;9(1-2):119–142. doi: 10.1007/978-1-4612-0475-6_8. [DOI] [PubMed] [Google Scholar]
  6. Bories D., Raynal M. C., Solomon D. H., Darzynkiewicz Z., Cayre Y. E. Down-regulation of a serine protease, myeloblastin, causes growth arrest and differentiation of promyelocytic leukemia cells. Cell. 1989 Dec 22;59(6):959–968. doi: 10.1016/0092-8674(89)90752-6. [DOI] [PubMed] [Google Scholar]
  7. Cai T. Q., Law S. K., Zhao H. R., Wright S. D. Reversible inactivation of purified leukocyte integrin CR3 (CD11b/CD18, alpha m beta 2) by removal of divalent cations from a cryptic site. Cell Adhes Commun. 1995 Dec;3(5):399–406. doi: 10.3109/15419069509081294. [DOI] [PubMed] [Google Scholar]
  8. Cai T. Q., Wright S. D. Energetics of leukocyte integrin activation. J Biol Chem. 1995 Jun 16;270(24):14358–14365. doi: 10.1074/jbc.270.24.14358. [DOI] [PubMed] [Google Scholar]
  9. Campbell E. J. Human leukocyte elastase, cathepsin G, and lactoferrin: family of neutrophil granule glycoproteins that bind to an alveolar macrophage receptor. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6941–6945. doi: 10.1073/pnas.79.22.6941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Campbell E. J., Silverman E. K., Campbell M. A. Elastase and cathepsin G of human monocytes. Quantification of cellular content, release in response to stimuli, and heterogeneity in elastase-mediated proteolytic activity. J Immunol. 1989 Nov 1;143(9):2961–2968. [PubMed] [Google Scholar]
  11. Dana N., Styrt B., Griffin J. D., Todd R. F., 3rd, Klempner M. S., Arnaout M. A. Two functional domains in the phagocyte membrane glycoprotein Mo1 identified with monoclonal antibodies. J Immunol. 1986 Nov 15;137(10):3259–3263. [PubMed] [Google Scholar]
  12. Detmers P. A., Zhou D., Powell D., Lichenstein H., Kelley M., Pironkova R. Endotoxin receptors (CD14) are found with CD16 (Fc gamma RIII) in an intracellular compartment of neutrophils that contains alkaline phosphatase. J Immunol. 1995 Aug 15;155(4):2085–2095. [PubMed] [Google Scholar]
  13. Diamond M. S., Alon R., Parkos C. A., Quinn M. T., Springer T. A. Heparin is an adhesive ligand for the leukocyte integrin Mac-1 (CD11b/CD1). J Cell Biol. 1995 Sep;130(6):1473–1482. doi: 10.1083/jcb.130.6.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Diamond M. S., Springer T. A. A subpopulation of Mac-1 (CD11b/CD18) molecules mediates neutrophil adhesion to ICAM-1 and fibrinogen. J Cell Biol. 1993 Jan;120(2):545–556. doi: 10.1083/jcb.120.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Diamond M. S., Springer T. A. The dynamic regulation of integrin adhesiveness. Curr Biol. 1994 Jun 1;4(6):506–517. doi: 10.1016/s0960-9822(00)00111-1. [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. T-cell receptor cross-linking transiently stimulates adhesiveness through LFA-1. Nature. 1989 Oct 19;341(6243):619–624. doi: 10.1038/341619a0. [DOI] [PubMed] [Google Scholar]
  18. Elemer G. S., Edgington T. S. Monoclonal antibody to an activation neoepitope of alpha M beta 2 inhibits multiple alpha M beta 2 functions. J Immunol. 1994 Jun 15;152(12):5836–5844. [PubMed] [Google Scholar]
  19. Gabay J. E., Almeida R. P. Antibiotic peptides and serine protease homologs in human polymorphonuclear leukocytes: defensins and azurocidin. Curr Opin Immunol. 1993 Feb;5(1):97–102. doi: 10.1016/0952-7915(93)90087-9. [DOI] [PubMed] [Google Scholar]
  20. Heck L. W., Rostand K. S., Hunter F. A., Bhown A. Isolation, characterization, and amino-terminal amino acid sequence analysis of human neutrophil cathepsin G from normal donors. Anal Biochem. 1986 Oct;158(1):217–227. doi: 10.1016/0003-2697(86)90612-3. [DOI] [PubMed] [Google Scholar]
  21. Hermanowski-Vosatka A., Van Strijp J. A., Swiggard W. J., Wright S. D. Integrin modulating factor-1: a lipid that alters the function of leukocyte integrins. Cell. 1992 Jan 24;68(2):341–352. doi: 10.1016/0092-8674(92)90475-r. [DOI] [PubMed] [Google Scholar]
  22. Jenne D. E. Structure of the azurocidin, proteinase 3, and neutrophil elastase genes. Implications for inflammation and vasculitis. Am J Respir Crit Care Med. 1994 Dec;150(6 Pt 2):S147–S154. doi: 10.1164/ajrccm/150.6_Pt_2.S147. [DOI] [PubMed] [Google Scholar]
  23. Lo S. K., Detmers P. A., Levin S. M., Wright S. D. Transient adhesion of neutrophils to endothelium. J Exp Med. 1989 May 1;169(5):1779–1793. doi: 10.1084/jem.169.5.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Owen C. A., Campbell M. A., Sannes P. L., Boukedes S. S., Campbell E. J. Cell surface-bound elastase and cathepsin G on human neutrophils: a novel, non-oxidative mechanism by which neutrophils focus and preserve catalytic activity of serine proteinases. J Cell Biol. 1995 Nov;131(3):775–789. doi: 10.1083/jcb.131.3.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pereira H. A. CAP37, a neutrophil-derived multifunctional inflammatory mediator. J Leukoc Biol. 1995 Jun;57(6):805–812. doi: 10.1002/jlb.57.6.805. [DOI] [PubMed] [Google Scholar]
  26. Pohl J., Pereira H. A., Martin N. M., Spitznagel J. K. Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. FEBS Lett. 1990 Oct 15;272(1-2):200–204. doi: 10.1016/0014-5793(90)80484-z. [DOI] [PubMed] [Google Scholar]
  27. Pulford K. A., Erber W. N., Crick J. A., Olsson I., Micklem K. J., Gatter K. C., Mason D. Y. Use of monoclonal antibody against human neutrophil elastase in normal and leukaemic myeloid cells. J Clin Pathol. 1988 Aug;41(8):853–860. doi: 10.1136/jcp.41.8.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Robinson M. K., Andrew D., Rosen H., Brown D., Ortlepp S., Stephens P., Butcher E. C. Antibody against the Leu-CAM beta-chain (CD18) promotes both LFA-1- and CR3-dependent adhesion events. J Immunol. 1992 Feb 15;148(4):1080–1085. [PubMed] [Google Scholar]
  29. Sinha S., Watorek W., Karr S., Giles J., Bode W., Travis J. Primary structure of human neutrophil elastase. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2228–2232. doi: 10.1073/pnas.84.8.2228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smyth S. S., Joneckis C. C., Parise L. V. Regulation of vascular integrins. Blood. 1993 Jun 1;81(11):2827–2843. [PubMed] [Google Scholar]
  31. Tosi M. F., Zakem H., Berger M. Neutrophil elastase cleaves C3bi on opsonized pseudomonas as well as CR1 on neutrophils to create a functionally important opsonin receptor mismatch. J Clin Invest. 1990 Jul;86(1):300–308. doi: 10.1172/JCI114699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Van Voorhis W. C., Steinman R. M., Hair L. S., Luban J., Witmer M. D., Koide S., Cohn Z. A. Specific antimononuclear phagocyte monoclonal antibodies. Application to the purification of dendritic cells and the tissue localization of macrophages. J Exp Med. 1983 Jul 1;158(1):126–145. doi: 10.1084/jem.158.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Weiss S. J. Tissue destruction by neutrophils. N Engl J Med. 1989 Feb 9;320(6):365–376. doi: 10.1056/NEJM198902093200606. [DOI] [PubMed] [Google Scholar]
  34. Wolfsberg T. G., Primakoff P., Myles D. G., White J. M. ADAM, a novel family of membrane proteins containing A Disintegrin And Metalloprotease domain: multipotential functions in cell-cell and cell-matrix interactions. J Cell Biol. 1995 Oct;131(2):275–278. doi: 10.1083/jcb.131.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Woodman R. C., Reinhardt P. H., Kanwar S., Johnston F. L., Kubes P. Effects of human neutrophil elastase (HNE) on neutrophil function in vitro and in inflamed microvessels. Blood. 1993 Oct 1;82(7):2188–2195. [PubMed] [Google Scholar]
  36. Wright S. D., Meyer B. C. Phorbol esters cause sequential activation and deactivation of complement receptors on polymorphonuclear leukocytes. J Immunol. 1986 Mar 1;136(5):1759–1764. [PubMed] [Google Scholar]
  37. 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]
  38. Wright S. D., Weitz J. I., Huang A. J., Levin S. M., Silverstein S. C., Loike J. D. Complement receptor type three (CD11b/CD18) of human polymorphonuclear leukocytes recognizes fibrinogen. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7734–7738. doi: 10.1073/pnas.85.20.7734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Zimmerman B. J., Granger D. N. Reperfusion-induced leukocyte infiltration: role of elastase. Am J Physiol. 1990 Aug;259(2 Pt 2):H390–H394. doi: 10.1152/ajpheart.1990.259.2.H390. [DOI] [PubMed] [Google Scholar]
  40. van Kessel K. P., Park C. T., Wright S. D. A fluorescence microassay for the quantitation of integrin-mediated adhesion of neutrophil. J Immunol Methods. 1994 Jun 3;172(1):25–31. doi: 10.1016/0022-1759(94)90375-1. [DOI] [PubMed] [Google Scholar]

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