Skip to main content
Log in

Stimulated release of neutral proteinases elastase and cathepsin G from inflammatory rat polymorphonuclear leukocytes

  • Original Articles
  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

Rat leukocytes from inflammatory peritoneal exudates respond in vitro to a variety of chemotactic and phagocytic stimuli by releasing both elastase and cathepsin G neutral proteinase enzyme activities. PAF, FMLP, and PMA stimulated a rapid, cytochalasin B-dependent, dose-related release of both enzymes; however, leukotriene B4 was inactive. It was not possible to measure the activity of zymosan-activated serum on these cells as rat serum contains high levels of proteinase inhibitors. The calcium ionophore A23187 stimulated a dose-related, time-dependent, cytochalasin B-independent enzyme release. Concanavalin A stimulated a weak, nondose-related release of enzyme activity. Zymosan and serum-coated zymosan stimulated enzyme secretion which was markedly inhibited by the presence of cytochalasin B. These data indicate that release of azurophillic granule neutral proteinases from rat inflammatory leukocytes can be detected and quantitated in vitro. This model could provide a test system for monitoring the pattern and specificity of enzyme release from azurophil granules. The ability of a variety of stimuli to induce proteolytic enzyme release from inflammatory neutrophils may be of considerable relevance to chronic inflammatory diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Klebanoff, S. J., andR. A. Clark. 1978. The Neutrophil: Function and Clinical Disorders. North Holland, Amsterdam.

    Google Scholar 

  2. Hoffstein, S. T. 1980. Intra-and extracellular secretion from polymorphonuclear leukocytes. The cell biology of inflammation.Handb. Inflamm. 2:387–430.

    Google Scholar 

  3. Calamai, E. G., andJ. K. Spitznagel. 1982. Characterization of rat polymorphonuclear leukocyte subcellular granules.Lab. Invest. 46:597–604.

    Google Scholar 

  4. Varani, J., D. Ward, andK. J. Johnson. 1982. Nonspecific protease and elastase activities in rat leukocytes.Inflammation 6:177–187.

    Google Scholar 

  5. Lewis, A. J., R. P. Carlson, andJ. Chang. 1985. Experimental models of inflammation. The pharmacology of inflammation.Handb. Inflamm. 5:371–397.

    Google Scholar 

  6. Baici, A. 1980. Structure and breakdown of proteoglycans, collagen and elastin.In Proteinases in Tumor Invasion. P. Stäuli, A. J. Barrett, and A. Baici, editors. Raven Press, New York. 17–48.

    Google Scholar 

  7. Baici, A., P. Salgam, G. Cohen, K. Fehr, andA. Böni. 1982. Action of collagenase and elastase from human polymorphonuclear leukocytes on human articular cartilage.Rheumatol. Int. 2:11–16.

    Google Scholar 

  8. Cohen, A. B. 1983. Proteases and antiproteases in the lung.Am. Rev. Respir. Dis. (Suppl.) 127:S2-S58.

    Google Scholar 

  9. Barrett, A. J., andJ. Saklatvala. 1981. Proteinases in joint disease.In Textbook of Rheumatology. W. N. Kelley, E. D. Harris, Jr., S. Ruddy, and C. B. Sledge, editors. W. B. Saunders, Philadelphia. 195–209.

    Google Scholar 

  10. Showell, H. J., R. J. Freer, S. H. Zigmond, E. Schiffmann, S. Aswanikumar, B. Corcoran, andE. L. Becker. 1976. The structure-activity relations of synthetic peptides as chemotactic factors and inducers of lysosomal enzyme secretion for neutrophils.J. Exp. Med. 143:1154–1169.

    Google Scholar 

  11. Sklar, L. A., P. A. Hyslop, Z. G. Oades, G. M. Omann, A. J. Jesaitis, R. G. Painter, andC. G. Cochrane. 1985. Signal transduction and ligand-receptor dynamics in the human neutrophil.J. Biol. Chem. 260:11461–11467.

    Google Scholar 

  12. Goldstein, I., S. Hoffstein, J. Gallin, andG. Wiessmann. 1973. Mechanisms of lysosomal enzyme release from human leukocytes: Microtubule assembly and membrane fusion induced by a component of complement.Proc. Natl. Acad. Sci. U.S.A. 70:2916–2920.

    Google Scholar 

  13. Bray, M. A. 1983. Pharmacology and pathophysiology of leukotriene B4.Br. Med. Bull. 39:249–254.

    Google Scholar 

  14. Feinmark, S. J., J. A. Lindgren, H. E. Claesson, C. Malmsten, andB. Samuelsson. 1981. Stimulation of human leukocyte degranulation by leukotriene B4 and its w-oxidized metabolites.FEBS Lett. 136:141–144.

    Google Scholar 

  15. Shaw, J. O., R. N. Pinckard, K. S. Ferrigni, L. M. McManus, andD. J. Hanahan. 1981. Activation of human neutrophils with 1-O-hexadecyl/octadecyl-2-acetyl-sn-glyceryl-3-phosphorylcholine (platelet activating factor).J. Immunol. 127:1250–1255.

    Google Scholar 

  16. Showell, H. J., P. H. Naccache, P. Borgeat, S. Picard, P. Vallerand, E. L. Becker, andR. I. Sha'afi. 1982. Characterization of the secretory activity of leukotriene B4 toward rabbit neutrophils.J. Immunol. 128:811–816.

    Google Scholar 

  17. Zurier, R. B., S. Hoffstein, andG. Weissmann. 1973. Cytochalasin B: Effect on lysosomal enzyme release from human leukocytes.Proc. Natl. Acad. Sci. U.S.A. 70:844–848.

    Google Scholar 

  18. Henson, P. M., H. B. Johnson, andH. L. Spiegelberg. 1972. The release of granule enzymes from human neutrophils stimulated by aggregated immunoglobulins of different classes and subclasses.J. Immunol. 109:1182–1192.

    Google Scholar 

  19. Estensen, R. D., M. E. Reusch, M. L. Epstein, andH. R. Hill. 1976. Role of Ca2+ and Mg2+ in some human neutrophil functions as indicated by ionophore A23187.Infect. Immun. 13:45–151.

    Google Scholar 

  20. Hoffstein, S., andG. Weissmann. 1978, Microfilaments and microtubules in calcium ionophore-induced secretion of lysosomal enzymes from human polymorphonuclear leukocytes.J. Cell. Biol. 78:769–781.

    Google Scholar 

  21. Estensen, R. D., J. G. White, andB. Holmes. 1974. Specific degranulation of human polymorphonuclear leukocytes.Nature 248:347–348.

    Google Scholar 

  22. Virca, G. D., G. Metz, andH. P. Schnebli. 1984. Similarities between human and rat leukocyte elastase and cathepsin G.Eur. J. Biochem. 144:1–9.

    Google Scholar 

  23. Sklar, L. A., V. M. McNeil, A. J. Jesaitis, R. G. Painter, andC. G. Cochrane. 1982. A continuous, spectroscopic analysis of the kinetics of elastase secretion by neutrophils.J. Biol. Chem. 257:5471–5475.

    Google Scholar 

  24. Anderson, A. J., andC. Irwin. 1973. Some properties of neutral acting proteases and other degradative enzymes in rat leukocytes.Life Sci. 13:601–612.

    Google Scholar 

  25. Baggiolini, M. 1985. Phagocyte activation and its modulation by drugs. The pharmacology of inflammation.Handb. Inflamm. 5:117–139.

    Google Scholar 

  26. O'Flaherty, J. T., R. L. Wykle, C. E. McCall, T. B. Shewmake, C. J. Lees, andM. Thomas 1981. Desensitisation of the human neutrophil degranulation response: Studies with 5, 12-dihydroxy-6,8,10,14-eicosatetraenoic acid.Biochem. Biophys. Res. Commun. 101:1290–1296.

    Google Scholar 

  27. Palmer, R. M. J., andJ. A. Salmon 1983. Release of leukotriene B4 from human neutrophils and its relationship to degranulation induced by N-formyl-methionyl-leucyl-phenylalanine, serum-treated zymosan, and the ionophore A23187.Immunology 50:65–73.

    Google Scholar 

  28. Becker, E. L., H. J. Showell, P. M. Henson, andL. S. Hsu 1974. The ability of chemotactic factors to induce lysosomal enzyme release. 1. The characteristics of the release, the importance of surfaces, and the relation of enzyme release to chemotactic responsiveness.J. Immunol. 112:2047–2054.

    Google Scholar 

  29. Bentwood, B. J., andP. M. Henson. 1980. The sequential release of granule constituents from human neutrophils.J. Immunol. 124: 855–862.

    Google Scholar 

  30. Wright, D. G., D. A. Bralove, andJ. I. Gallin. 1977. The differential mobilisation of human neutrophil granules. Effects of phorbol myristate acetate and ionophore A23187.Am. J. Palhol. 87:273–281.

    Google Scholar 

  31. Yurewicz, E. C., andM. Zimmerman. 1977. Cytochalasin B-dependent release of azurophil granule enzymes from human polymorphonuclear leukocytes.Inflammation 2:259–264.

    Google Scholar 

  32. Abrams, W. R., L. W. Diamond, andA. B. Kane. 1983. A flow cytometric assay of neutrophil degranulation.J. Histochem. Cytochem. 31:737–744.

    Google Scholar 

  33. Blackwood, C. E., J. E. Moret, S. Keller, J. A. Fierer, andI. Mandl. 1974. Alphai- antitrypsin concentration in serum of laboratory animals. Changes following proteolytic enzyme injection.J. Lab. Clin. Med. 84:813–822.

    Google Scholar 

  34. Hawkins, D. 1974. Neutrophilic leukocytes in immunologic reactions in vitro. III Pharmacologic modulation of lysosomal constituent release.Clin. Immunol. Immunopathol. 2:141–152.

    Google Scholar 

  35. Hoffstein. S., R. Soberman, I. Goldstein, andG. Weissmann. 1976. Concanavalin A induces microtubule assembly and specific granule discharge in human polymorphonuclear leukocytes.J. Cell. Biol. 68:781–787.

    Google Scholar 

  36. Smith, R. J., andS. S. Iden. 1980. Properties of concanavalin A-elicited granule exocytosis from human polymorphonuclear neutrophils.Inflammation 4:343–358.

    Google Scholar 

  37. Hoffstem, S., R. B. Zurier, andG. Weissmann. 1974. Mechanisms of lysosomal enzyme release from human leukocytes. III Quantitative morphologic evidence for an effect of cyclic nucleotides and colchicine on degranulation.Clin. Immunol. Immunopathol. 3:201–217.

    Google Scholar 

  38. Weissmann, G., R. B. Zurier, andS. Hoffstein. 1973. Leukocytes as secretory organs in inflammation.Agents Actions 3:370–379.

    Google Scholar 

  39. Hadler, N. M., J. K. Spitznagel, andR. J. Quinet. 1979. Lysosomal enzymes in inflammatory synovial effussions.J. Immunol. 123:572–577.

    Google Scholar 

  40. Saklatvala, J., andA. J. Barrett. 1980. Identification of proteinases in rheumatoid synovium. Detection of leukocyte elastase, cathepsin G, and another serine proteinase.Biochim. Biophys. Acta 615:167–177.

    Google Scholar 

  41. Virca, G. D., R. K. Mallya, M. B. Pepys, andH. P. Schnebli. 1984. Quantitaton of human leukocyte elastase, cathepsin G, α2-macroglobulin and α1-proteinase inhibitor in osteoarthrosis and rheumatoid arthritis synovial fluids.In Proteases. W. H. Horl and A. Heidland, editors. Plenum Publishing, New York, 345–353.

    Google Scholar 

  42. Postma, D. S., B. Folkers, H. J. Sluiter, andH. F. Kauffman. 1986. Discrepancies in release of elastase from human peripheral polymorphonuclear leukocytes.Eur. J. Resp. Dis. (Suppl. No. 146)69:217–224.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bray, M.A., McKearn-Smith, C., Metz-Virca, G. et al. Stimulated release of neutral proteinases elastase and cathepsin G from inflammatory rat polymorphonuclear leukocytes. Inflammation 11, 23–37 (1987). https://doi.org/10.1007/BF00917769

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00917769

Keywords

Navigation