Skip to main content
SpringerLink
Log in

Interference of cartilage surface with interaction of granulocyte elastase with α1-proteinase inhibitor

An in vitro model of enzyme inhibition in the joint space

  • Originals
  • Published:
Rheumatology International Aims and scope Submit manuscript

Summary

Synovial fluids of patients suffering from rheumatoid arthritis contain elevated levels of granulocyte (PMN) elastase in complex with alpha 1-proteinase inhibitor (alpha 1-PI), whereas free-elastase activity is usually not detectable. This absence of free enzymatic activity in joint effusions has cast some doubt on the pathophysiological relevance of PMN elastase in inflammatory joint destruction. Our in vitro experiments using bovine nasal cartilage demonstrate that incubation with elastase and alpha 1-PI in equimolar concentrations to or even in excess of the serum proteinase inhibitor resulted in significant tissue destruction as assessed by histological staining for proteoglycans, release of uronic acid from the matrix and loss of mechanical stability. Though in the supernatants containing alpha 1-PI, free-elastase activity was not detectable, immunofluorescent staining for elastase evidenced penetration of the enzyme into the matrix. Simultaneous measurements of the incubation media employing a sandwich enzyme-linked immunoadsorption assay (ELISA) revealed PMN elastase in complex with alpha 1-PI but without correlation to the parameters of tissue degradation. In comparison with the results obtained using the chromogenic substrate Suc-Ala-Ala-Ala-pNA (SAPA) for titration of alpha 1-PI against elastase, the employment of cartilage matrix showed that a fourfold increase in inhibitor concentration was necessary to achieve 100% enzyme inhibition. Hence, cartilage surface obviously interferes with the interaction between alpha 1-PI and elastase. Measurements of elastase-inhibitor concentrations or free enzymatic activity in synovial fluid seem to have limited value in predicting cartilage destruction.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Barret AJ (1978) The possible role of neutrophil proteinases in damage to articular cartilage. Agents Actions 8:11–18

    Google Scholar 

  2. Gosh P (1978) Lysosomal enzymes in joint disease. Aust NZ J Med 8 (suppl 1):12–15

    Google Scholar 

  3. Hadler NM, Johnson AM, Spitznagel JK, Quinet RJ (1981) Protease inhibitors in inflammatory synovial effusions. Ann Rheum Dis 40:55–59

    Google Scholar 

  4. Dingle JT (1979) Recent studies on the control of joint damage: The contribution of the Strangeway Research Laboratory, Heberden Oration 1978. Ann Rheum Dis 38:201–214

    Google Scholar 

  5. Starkey PM, Barret AJ, Burleigh MC (1977) The degradation of articular collagen by neutrophil proteinases. Biochim Biophys Acta 483:386–397

    Google Scholar 

  6. Keiser H, Greenwald RA, Feinstein G, Janoff A (1976) Degradation of cartilage proteoglycan by human leukocyte granule neutral protease — A model of joint injury. J Clin Invest 57:625–632

    Google Scholar 

  7. Kleesiek K, Reinards R, Brackerts D, Neumann S, Lang H, Greiling H (1986) Granulocyte elastase as a new biochemical marker in the diagnosis of chronic joint diseases. Rheumatol Int 6:161–170

    Google Scholar 

  8. Virca GD, Mallya RK, Pepys MB, Schnebli HP (1984) Quantitation of human leukocyte elastase, cathepsin G, alpha 2-macroglobulin and alpha 1-proteinase inhibitor in osteoarthrosis and rheumatiod arthritis synovial fluids. Adv Exp Biol 167:345–353

    Google Scholar 

  9. Cohen G, Fehr K, Wagenhäuser FJ (1983) Leukocyte elastase and free collagenase activity in synovial effusions: relation to numbers of polymorphonuclear leukocytes. Rheumatol Int 3:89–95

    Google Scholar 

  10. Burkhardt H, Schwingel ML, Menninger H, Macartney HW, Tschesche H (1986) Oxygen radicals as effectors of cartilage destruction. Arthritis Rheum 29:379–387

    Google Scholar 

  11. Barthold PM, Page RC (1985) A microdetermination method for assaying glycosaminoglycans and proteoglycans. Anal Biochem 150:320–324

    Google Scholar 

  12. Stegemann H (1958) Mikrobestimmung von Hydroxyprolin mit Chloramin-T und p-Dimethylaminobenzaldehyd. Z Physiol Chem 311:41–45

    Google Scholar 

  13. Nakajima K, Powers JC, Castillo MJ, Ashe BM, Zimmermann M (1979) Mapping the extended substrate binding site of cathepsin G and human leukocyte elastase. J Biol Chem 245:4027–4031

    Google Scholar 

  14. Mowry RW (1956) Alcian blue techniques for the histochemical study of acidic carbohydrates. J Histochem Cytochem 4:407

    Google Scholar 

  15. Menninger H, Putzier R, Mohr W, Wessinghage D, Tillmann K (1980) Granulocyte elastase at the site of cartilage erosion by rheumatoid synovial tissue. Z Rheumatol 39:145–156

    Google Scholar 

  16. Menninger H, Burkhardt H, Röske W, Ehlebracht W, Hering B, Gurr E, Mohr W, Mierau HD (1981) Lysosomal elastase: Effect on mechanical and biochemical properties of normal cartilage, inhibition by polysulfonated glycosaminoglycan and binding to chondrocytes. Rheumatol Int 1:73–82

    Google Scholar 

  17. Engelbrecht S, Pieper E, Macartney HW, Rautenberg W, Wenzel HR, Tschesche H (1981) Separation of the human leukocyte enzymes alanine aminopeptidase, cathepsin G, collagenase, elastase and myeloperoxidase. Hoppe-Seyler's Z Physiol Chem 363:305–315

    Google Scholar 

  18. Tschesche H, Engelbrecht S, Wenzel HR (1979) In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 5. Verlag Chemie, Weinheim, pp 176–183

    Google Scholar 

  19. Maroudas A (1973) Physicochemical properties of articular cartilage. In: Freeman MAR (ed) Adult articular cartilage. Pitmann, London, pp 131–170

    Google Scholar 

  20. Janoff A (1973) Purification of human granulocyte elastase by affinity chromatography. Lab Invest 29:458–464

    Google Scholar 

  21. Stockley RA, Afford SC (1984) The effect of leukocyte elastase on the immunoelectrophoretic behavior of alpha 1-antitrypsin. Clin Sci 66:217–224

    Google Scholar 

  22. Hornebeck W, Soleilhac JM, Velebuy V, Robert L (1985) On the influence of the substrate (elastin) in elastase-alpha 1-antitrypsin interactions. Pathol Biol 33:281–285

    Google Scholar 

  23. Pritchard MH (1984) Synovial protease inhibitor ratios in erosive and nonerosive arthropaties. Ann Rheum Dis 43:50–55

    Google Scholar 

  24. Kimura H, Tateishi H, Ziff M (1977) Surface ultrastructure of rheumatoid articular cartilage. Arthritis Rheum 20:1085–1094

    Google Scholar 

  25. Taillard W, Lagier R (1969) Chondromalacia and arthritis of rheumatoid type. In: Hijmans W, Paul WD, Herschel H (eds) Early synovectomy in rheumatoid arthritis. Excerpta Medica Foundation, Amsterdam, pp 178–184

    Google Scholar 

  26. Drommer W (1982) Erreger bestimmen im Zusammenspiel mit Leukozyten und Fibrin die initiale Phase der Gelenkdestruktion. In: Otte P (Hrsg) Gelenkdestruktion bei Polyarthritis. Steinkopff, Darmstadt, pp 15–21

    Google Scholar 

  27. Velvart M, Fehr K, Baici A, Sommermeyer G, Knöpfel M, Cancer M, Salgam P, Böni A (1981) Degradation in vivo of articular cartilage in rheumatoid arthritis by leukocyte elastase from polymorphonuclear leukocytes. Rheumatol Int 1:121–130

    Google Scholar 

  28. Matheson NR, Wang PS, Travis J (1979) Enzymatic inactivation of human alpha 1-proteinase inhibitor by neutrophil myeloperoxidase. Biochim Biophys Res Comm 88:402–409

    Google Scholar 

  29. Johnson D, Travis J (1979) The oxidative inactivation of human alpha 1-proteinase inhibitor: further evidence for methionine at the reactive center. J Biol Chem 254:4022–4026

    Google Scholar 

  30. Carp H, Janoff A (1979) In vitro suppression of serum elastase inhibitory capacity by reactive oxygen species generated by phagocytosing polymorphonuclear leukocytes. J Clin Invest 63:793–797

    Google Scholar 

  31. Henson PM (1971) The immunologic release of constituents from neutrophil leukocytes. II. Mechanism of release during phagocytosis and adherence to nonphagocytosable surfaces. J Immunol 107:1547–1557

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Abt. Krankheiten der Bewegungsorgane und des Stoffwechsels, Medizinische Hochschule Hannover, Konstanty-Gutschow-Strasse 8, D-3000, Hannover 61, Federal Republic of Germany

    H. Burkhardt & E. Rehkopf

  2. Fakultät für Chemie, Lehrstuhl für Biochemie, Universität Bielefeld, Universitätsstrasse, D-4800, Bielefeld 1, Federal Republic of Germany

    M. Kasten & S. Rauls

Authors
  1. H. Burkhardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Kasten
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Rauls
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E. Rehkopf
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Burkhardt, H., Kasten, M., Rauls, S. et al. Interference of cartilage surface with interaction of granulocyte elastase with α1-proteinase inhibitor. Rheumatol Int 7, 133–138 (1987). https://doi.org/10.1007/BF00270466

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00270466

Key words

Search

Navigation