Journal of Clinical Immunology

, Volume 15, Issue 1, pp 27–34 | Cite as

Presence, activities, and molecular forms of cathepsin G, elastase,α1-antitrypsin, andα1-antichymotrypsin in bronchiectasis

  • Ruth Sepper
  • Yrjö T. Konttinen
  • Tuula Ingman
  • Timo Sorsa
Original Articles


The presence, activities, and molecular forms of the serine proteinases, elastase, and cathepsin G, and their endogenous inhibitors,α1-antitrypsin andα1-antichymotrypsin, were investigated in bronchoalveolar lavage (BAL) fluid of bronchiectasis patients divided into mild, moderate, and severe disease subgroups and compared to BAL fluid from healthy controls. Immunochemical characterization and quantitation were performed by Western immunoblot. The activities of elastase and cathepsin G were recorded spectrophotometrically using synthetic substrates. The results showed a significant difference in elastase and cathepsin G activities in BAL fluid of the three subgroups, revealing the following data—mild subgroup, 0.21±0.09 mU/g and 57.35±20.9 U/g; moderate subgroup, 1.87±1.12 mU/g and 89.24±31.4 U/g; and severe subgroup, 2.64±1.63 mU/g and 139.18±58.3 U/g, respectively—compared to those of the healthy control group, 0.09±0.03 mU/g and 50.96±16.5 U/g. Evidently, the protective shield of plasma-derived antiproteinases was sufficient in healthy subjects and, also, in mild cases of bronchiectasis, but not in cases of severe and moderate forms of bronchiectasis, in which free and catalytically active elastase and cathepsin G were detected. The serine proteinases inhibitors (serpins),α1-antitrypsin andα1-antichymotrypsin, have evidently been oxidized and/or proteolytically cleaved in the cases of moderate and severe bronchiectasis. The results indicate that insufficient endogenous downregulation of catalytically active elastase and cathepsin G in BALF leads to tissue injury, resulting in alterative and deformative processes in the bronchiectasis lung.

Key words

Bronchiectasis bronchoalveolar lavage fluid polymorphonuclear leukocyte elastase cathepsin G α1-antitrypsin α1-antichymotrypsin 

Abbreviations used


bronchoalveolar lavage


bronchoalveolar lavage fluid








extracellular matrix


computerized tomography


polymorphonuclear leukocyte


nitrocellulose membrane


sodium dodecyl sulfatepolyacrylamide gel electrophoresis


Tris-buffered saline


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Turino GM: Natural history and clinical management of emphysema in patients with and without alpha 1-antitrypsin inhibitor deficiency. Ann NY Acad Sci 624:18–29, 1991Google Scholar
  2. 2.
    Janoff A: Elastases and emphysema. Current assessment of the protease-antiprotease hypothesis. Am Rev Resp Dis 132:417–433, 1985Google Scholar
  3. 3.
    Stockley RA: Proteolytic enzymes, their inhibitors and lung diseases. Clin Sci 64:119–126, 1983Google Scholar
  4. 4.
    Gadek JE, Pacht ER: The protease-antiprotease balance within the human lung: Implications for the pathogenesis of emphysema. Lung 168 (Suppl):552–564, 1990Google Scholar
  5. 5.
    Weiss SJ: Tissue destruction by neutrophils. N Engl J Med 320:365–376, 1989Google Scholar
  6. 6.
    Venge P, Rak S, Steinholtz L, Hakansson L, Lindblad G: Neutrophil function in chronic bronchitis. Eur Resp J 4:536–543, 1991Google Scholar
  7. 7.
    Hill SL, Burnett D, Hewetson KA, Stockley RA: The response of patients with purulent bronchiectasis to antibiotics for four months. Q J Med 66:163–173, 1988Google Scholar
  8. 8.
    Stockley RA: Bronchiectasis—New therapeutic approaches based on pathogenesis. Clin Chest Med 8:481–494, 1987Google Scholar
  9. 9.
    Campbell EJ, Senior RM, Welgus HG: Extracellular matrix injury during lung inflammation. Chest 92:161–167, 1987Google Scholar
  10. 10.
    Lloberes P, Montserrat E, Montserrat JM, Picado C: Sputum sol phase proteins and elastase activity in patients with clinically stable bronchiectasis. Thorax 47:88–92, 1992Google Scholar
  11. 11.
    Fahy JV, Schuster A, Ueki I, Boushey HA, Nadel JA: Mucus hypersecretion in bronchiectasis. The role of neutrophil proteases. Am Rev Resp Dis 146:1430–1433, 1992Google Scholar
  12. 12.
    Tetley TD: New perspectives on basic mechanisms in lung disease. 6. Proteinase imbalance: Its role in lung disease. Thorax 48:560–565, 1993Google Scholar
  13. 13.
    Stockley RA, Hill SL, Morrison HM, Starkie CM: Elastolytic activity of sputum and its relation to purulence and to lung function in patients with bronchiectasis. Thorax 39:408–413, 1984Google Scholar
  14. 14.
    Stockley RA, Burnett D: Alpha-1-antitrypsin and leukocyte elastase in infected and noninfected sputum. Am Rev Resp Dis 120:1081–1086, 1979Google Scholar
  15. 15.
    Rickard KA, Taylor J, Rennard SI: Observations of development of resistance to detachment of cultured bovine bronchial epithelial cells in response to protease treatment. Am J Resp Cell Mol Biol 6:414–420, 1992Google Scholar
  16. 16.
    Sommerhoff CP, Nadel JA, Basbaum CB, Caughey GH: Neutrophil elastase and cathepsin G stimulate secretion from cultured bovine airway gland serous cells. J Clin Invest 85:682–689, 1990Google Scholar
  17. 17.
    Barrett AJ: Leukocyte elastase. Methods Enzymol 80 Pt C:581–588, 1981Google Scholar
  18. 18.
    Travis J, Dubin A, Potempa J, Watorek W, Kurdowska A: Neutrophil proteinases. Caution signs in designing inhibitors against enzymes with possible multiple functions. Ann NY Acad Sci 624:81–86, 1991Google Scholar
  19. 19.
    Roughley PJ: The degradation of cartilage proteoglycans by tissue proteinases. Proteoglycan heterogeneity and the pathway of proteolytic degradation. Biochem J 167:639–646, 1977Google Scholar
  20. 20.
    Starkey PM, Barrett AJ, Burleigh MC: The degradation of articular collagen by neutrophil proteinases. Biochim Biophys Acta 483:386–397, 1977Google Scholar
  21. 21.
    Gadek JE, Fells GA, Wright DG, Crystal RG: Human neutrophil elastase functions as a type III collagen “collagenase”. Biochem Biophys Res Commun 95:1815–1822, 1980Google Scholar
  22. 22.
    Heck LW, Blackburn WD, Irwin MH, Abrahamson DR: Degradation of basement membrane laminin by human neutrophil elastase and cathepsin G. Am J Pathol 136:1267–1274, 1990Google Scholar
  23. 23.
    Mainardi CL, Dixit SN, Kang AH: Degradation of type IV (basement membrane) collagen by a proteinase isolated from human polymorphonuclear leukocyte granules. J Biol Chem 255:5435–5441, 1980Google Scholar
  24. 24.
    Boudier C, Holle C, Bieth JG: Stimulation of the elastolytic activity of leukocyte elastase by leukocyte cathepsin G. J Biol Chem 256:10256–10258, 1981Google Scholar
  25. 25.
    Okada Y, Morodomi T, Enghild JJ, Suzuki K, Yasui A, Nakanishi I, Salvesen G, Nagase H: Matrix metalloproteinase 2 from human rheumatoid synovial fibroblasts. Purification and activation of the precursor and enzymic properties. Eur J Biochem 194:721–730, 1990Google Scholar
  26. 26.
    Chatham WW, Blackburn WDJ, Heck LW: Additive enhancement of neutrophil collagenase activity by HOCl and cathepsin G. Biochem Biophys Res Commun 184:560–567, 1992Google Scholar
  27. 27.
    Capodici C, Muthukumaran G, Amoruso MA, Berg RG: Activation of neutrophil collagenase by cathepsin G. Inflammation 13:245–258, 1989Google Scholar
  28. 28.
    O'Connor CM, Gaffney K, Keane J, Southey A, Byrne N, O'Mahoney S, Fitzgerald MX:α-Proteinase inhibitor, elastase activity, and lung disease severity in cystic fibrosis. Am Rev Resp Dis 148:1665–1670, 1993Google Scholar
  29. 29.
    Meyer KC, Lewandoski JR, Zimmerman JJ, Nunley D, Calhoun WJ, Dopico GA: Human neutrophil elastase and elastase/alpha1-antiprotease complex in cystic fibrosis. Am Rev Resp Dis 144:580–585, 1991Google Scholar
  30. 30.
    Chandra T, Stackhouse R, Kidd VJ, Robson KJ, Woo SL: Sequence homology between human alpha 1-antichymotrypsin, alpha 1-antitrypsin, and antithrombin III. Biochemistry 22:5055–5061, 1983Google Scholar
  31. 31.
    Stockley RA, Burnett D: Serum derived protease inhibitors and leucocyte elastase in sputum and the effect of infection. Bull Eur Physiopathol Resp 16 (Suppl):261–272, 1980Google Scholar
  32. 32.
    Haslam PL: Bronchoalveolar lavage. Semin Resp Med 6:55–70, 1984Google Scholar
  33. 33.
    Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal Biochem 72:248–254, 1976Google Scholar
  34. 34.
    Nakajima MC, Powers JC, Ashe BM, Zimmerman M: Mapping of the extended substrate binding site of cathepsin G and human leukocyte elastase. J Biol Chem 254:4027–4032, 1979Google Scholar
  35. 35.
    Bieth J, Spiess B, Wermuth CG: The synthesis and analytical use of a highly sensitive and convenient substrate of elastase. Biochem Med 11:350–357, 1974Google Scholar
  36. 36.
    Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, 1970Google Scholar
  37. 37.
    Towbin H, Staehlin T, Gorden J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354, 1974Google Scholar
  38. 38.
    Hsu SM, Raine L, Fanger H: The use of antiavidin antibody and avidin-biotin-peroxidase complex in immunoperoxidase technics. Am J Clin Pathol 75:816–821, 1981Google Scholar
  39. 39.
    Renesto P, Chignard M: Tumor necrosis factor-alpha enhances platelet activation via cathepsin G released from neutrophils. J Immunol 146:2305–2309, 1991Google Scholar
  40. 40.
    Baugh RJ, Travis J: Human leukocyte granule elastase: Rapid isolation and characterization. Biochemistry 15:836–841, 1976Google Scholar
  41. 41.
    Wallaert B, Gressier B, Aerts C, Mizon C, Voisin C, Mizon J: Oxidative inactivation of alpha 1-proteinase inhibitor by alveolar macrophages from healthy smokers requires the presence of myeloperoxidase. Am J Resp Cell Mol Biol 5:437–444, 1991Google Scholar
  42. 42.
    Ossanna PJ, Test ST, Matheson NR, Regiani S, Weiss SJ: Oxidative regulation of neutrophil elastase-alpha-1-proteinase inhibitor interactions. J Clin Invest 77:1939–1951, 1986Google Scholar
  43. 43.
    Carp H, Janoff A: In vitro suppression of serum elastase-inhibitory capacity by reactive oxygen species generated by phagocytosing polymorphonuclear leukocytes. J Clin Invest 63:793–797, 1979Google Scholar
  44. 44.
    Carp H, Janoff A: Potential mediator of inflammation. Phagocytederived oxidants suppress the elastase-inhibitory capacity of alpha 1-proteinase inhibitor in vitro. J Clin Invest 66:987–995, 1980Google Scholar
  45. 45.
    Matheson NR, Wong PS, Schuyler M, Travis J: Interaction of human alpha-1-proteinase inhibitor with neutrophil myeloperoxidase. Biochemistry 20:331–336, 1981Google Scholar
  46. 46.
    Desrochers PE, Jeffrey JJ, Weiss SJ: Interstitial collagenase (matrix metallo-proteinase-1) expresses serpinase activity. J Clin Invest 87:2258–2265, 1991Google Scholar
  47. 47.
    Desrochers PE, Mookhtiar K, Van Wart HE, Hasty KA, Weiss SJ: Proteolytic inactivation ofα 1-proteinase inhibitor andα 1-antichymotrypsin by oxidatively activated human neutrophil metalloproteinases. J Biol Chem 267:5005–5012, 1992Google Scholar
  48. 48.
    Havemann K, Gramse M: Physiology and pathophysiology of neutral proteinases of human granulocytes. Adv Exp Med Biol 167:1–20, 1984Google Scholar
  49. 49.
    Beatty K, Bieth J, Travis J: Kinetics of association of serine proteinases with native and oxidized alpha-1-proteinase inhibitor and alpha-1-antichymotrypsin. J Biol Chem 255:3931–3934, 1980Google Scholar
  50. 50.
    Beatty K, Matheson N, Travis J: Kinetic and chemical evidence for the inability of oxidized alpha 1-proteinase inhibitor to protect lung elastin from elastolytic degradation. Hoppe Seylers Z Physiol Chem 365:731–736, 1984Google Scholar
  51. 51.
    Reilly CF, Travis J: The degradation of human lung elastin by neutrophil proteinases. Biochim Biophys Acta 621:147–157, 1980Google Scholar
  52. 52.
    Hornebeck W, Schnebli HP: Effect of different elastase inhibitors on leukocyte elastase pre-adsorbed to elastin. Hoppe Seylers Z Physiol Chem 363:455–458, 1982Google Scholar
  53. 53.
    Moroi M, Yamasaki M: Mechanism of interaction of bovine trypsin with human alpha 1-antitrypsin. Biochim Biophys Acta 359:130–141, 1974Google Scholar
  54. 54.
    Travis J, Bowen J, Baugh R: Human alpha-1-antichymotrypsin: Interaction with chymotrypsin-like proteinases. Biochemistry 17:5651–5656, 1978Google Scholar
  55. 55.
    Catanese J, Kress LF: Enzymatic inactivation of human plasma C1-inhibitor and alpha 1-antichymotrypsin by Pseudomonas aeruginosa proteinase and elastase. Biochim Biophys Acta 789:37–43, 1984Google Scholar
  56. 56.
    Berman G, Afford SC, Burnett D, Stockley RA: Alpha 1-Antichymotrypsin in lung secretions is not an effective proteinase inhibitor. J Biol Chem 25:14095–14099, 1986Google Scholar
  57. 57.
    Caughey GH: Roles of mast cell tryptase and chymase in airway function. Am J Physiol 257:L39-L46, 1989Google Scholar
  58. 58.
    Powers JC, Tanaka T, Harper JW, Minematsu Y, Barker L, Lincoln D, Crumley KV, Fraki JE, Schechter NM, Lazarus GG, Nakajima K, Nakashino K, Neurath H, Woodbury RG: Mammalian chymotrypsin-like enzymes. Comparative reactivities of rat mast cell proteases, human and dog skin chymases, and human cathepsin G with peptide 4-nitroanilide substrates and with peptide choloromethyl ketone and sulfonyl fluoride inhibitors. Biochemistry 24:2048–2058, 1985Google Scholar

Copyright information

© Plenum Publishing Corporation 1995

Authors and Affiliations

  • Ruth Sepper
    • 1
    • 2
  • Yrjö T. Konttinen
    • 1
  • Tuula Ingman
    • 4
  • Timo Sorsa
    • 3
  1. 1.Department of Anatomy, Laboratory of Molecular BiologyUniversity of HelsinkiHelsinkiFinland
  2. 2.Lung ClinicUniversity of TartuTartuEstonia
  3. 3.Department of PeriodontologyUniversity of HelsinkiHelsinkiFinland
  4. 4.Department of Medical ChemistryUniversity of HelsinkiHelsinkiFinland

Personalised recommendations