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.
Similar content being viewed by others
References
Klebanoff, S. J., andR. A. Clark. 1978. The Neutrophil: Function and Clinical Disorders. North Holland, Amsterdam.
Hoffstein, S. T. 1980. Intra-and extracellular secretion from polymorphonuclear leukocytes. The cell biology of inflammation.Handb. Inflamm. 2:387–430.
Calamai, E. G., andJ. K. Spitznagel. 1982. Characterization of rat polymorphonuclear leukocyte subcellular granules.Lab. Invest. 46:597–604.
Varani, J., D. Ward, andK. J. Johnson. 1982. Nonspecific protease and elastase activities in rat leukocytes.Inflammation 6:177–187.
Lewis, A. J., R. P. Carlson, andJ. Chang. 1985. Experimental models of inflammation. The pharmacology of inflammation.Handb. Inflamm. 5:371–397.
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.
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.
Cohen, A. B. 1983. Proteases and antiproteases in the lung.Am. Rev. Respir. Dis. (Suppl.) 127:S2-S58.
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.
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.
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.
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.
Bray, M. A. 1983. Pharmacology and pathophysiology of leukotriene B4.Br. Med. Bull. 39:249–254.
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.
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.
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.
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.
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.
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.
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.
Estensen, R. D., J. G. White, andB. Holmes. 1974. Specific degranulation of human polymorphonuclear leukocytes.Nature 248:347–348.
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.
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.
Anderson, A. J., andC. Irwin. 1973. Some properties of neutral acting proteases and other degradative enzymes in rat leukocytes.Life Sci. 13:601–612.
Baggiolini, M. 1985. Phagocyte activation and its modulation by drugs. The pharmacology of inflammation.Handb. Inflamm. 5:117–139.
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.
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.
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.
Bentwood, B. J., andP. M. Henson. 1980. The sequential release of granule constituents from human neutrophils.J. Immunol. 124: 855–862.
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.
Yurewicz, E. C., andM. Zimmerman. 1977. Cytochalasin B-dependent release of azurophil granule enzymes from human polymorphonuclear leukocytes.Inflammation 2:259–264.
Abrams, W. R., L. W. Diamond, andA. B. Kane. 1983. A flow cytometric assay of neutrophil degranulation.J. Histochem. Cytochem. 31:737–744.
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.
Hawkins, D. 1974. Neutrophilic leukocytes in immunologic reactions in vitro. III Pharmacologic modulation of lysosomal constituent release.Clin. Immunol. Immunopathol. 2:141–152.
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.
Smith, R. J., andS. S. Iden. 1980. Properties of concanavalin A-elicited granule exocytosis from human polymorphonuclear neutrophils.Inflammation 4:343–358.
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.
Weissmann, G., R. B. Zurier, andS. Hoffstein. 1973. Leukocytes as secretory organs in inflammation.Agents Actions 3:370–379.
Hadler, N. M., J. K. Spitznagel, andR. J. Quinet. 1979. Lysosomal enzymes in inflammatory synovial effussions.J. Immunol. 123:572–577.
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.
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.
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.
Author information
Authors and Affiliations
Rights 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
Issue Date:
DOI: https://doi.org/10.1007/BF00917769