Skip to main content
SpringerLink
Log in

Elastase activity in bronchoalveolar lavage fluid from oxygen-exposed,Pseudomonas-infected baboons

  • Published:
Lung Aims and scope Submit manuscript

Abstract

The adult respiratory distress syndrome is a major cause of morbidity and mortality in critical care patients. Lung injury in this syndrome is frequently associated with lung infection. The combined insults result in an influx of neutrophils and damage to the pulmonary epithelium. We investigated whether active neutrophil elastolytic activity was present in the bronchoalveolar fluid in baboons with mild or moderate hyperoxic lung injury and infection. Group A (N=7) was exposed for 6 days to FIO2=0.8 and then inoculated by intratracheal bolus withPseudomonas aeruginosa strain DGI-R130 (PA); the FIO2 was reduced to 0.5. Group B (N=6) was exposed to similar concentrations of inspired oxygen but inoculated with buffered saline. Antibiotics included parenteral penicillin and topical gentamicin and polymyxin B. All 3 were given continuously in group B but stopped 24 h prior to PA inoculation in group A. Bronchoalveolar lavage fluid was collected 1 week before oxygen administration, when the FIO2 was reduced (day 6 or 7) and prior to necropsy (day 11). Hemodynamic, pulmonary function, microbiological, and biochemical variables were studied. Injured, infected animals (group A) had significant elevations of mean pulmonary artery pressure and decreases in total lung capacity and PaO2 compared both to baseline and to group B at day 11. At autopsy, group A had significant increases of bronchoalveolar lavage fluid (BALF) neutrophils and bacterial pathogens. Elastase levels in BALF (equal to 0 at baseline) rose to 136±98 ng/ml in group A vs. 6±14 ng/ml in group B. The elastase was inhibited by inhibitors of serine proteases including ones specific for neutrophil elastase. On Sephacryl S-300 chromatography the elastase activity eluted near humanα 2-macroglobulin and separated from other proteolytic activity. These studies demonstrate a significant level of elastase in BALF from injured, infected baboons compared to injured, uninfected animals.

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. Abrams WR, Fein AM, Kucich U, Kueppers F, Yamada H, Kuzmowycz T, Morgan L, Lippmann M, Goldberg SK, Weinbaum G (1984) Proteinase inhibitory function in inflammatory lung disease. I. Acute bacterial pneumonia. Am Rev Respir Dis 129:735–741

    PubMed  CAS  Google Scholar 

  2. Beatty K, Bieth J, Travis J (1980) Kinetics of association of serine proteinases with native and oxidized α1-proteinase inhibitor and α1-antichymotrypsin. J Biol Chem 255:3931–3934

    PubMed  CAS  Google Scholar 

  3. Beatty K, Robertie P, Senior RM, Travis J (1982) Determination of oxidized alpha-1-proteinase inhibitor in serum. J Lab Clin Med 100:186–192

    PubMed  CAS  Google Scholar 

  4. Bell RC, Coalson JJ, Smith JD, Johanson WG Jr (1983) Multiple organ system failure and infection in adult respiratory distress syndrome. Ann Intern Med 99:293–298

    PubMed  CAS  Google Scholar 

  5. Campbell GD, Coalson JJ, Johanson WG Jr (1984) The effect of bacterial superinfection on lung function after diffuse alveolar damage. Am Rev Respir Dis 129:974–978

    PubMed  CAS  Google Scholar 

  6. Castillo MJ, Nakajima K, Zimmerman M, Powers JC (1979) Sensitive substrates for human leukocyte and porcine pancreatic elastase: a study of the merits of various chromophoric and fluorogenic leaving groups in assays for serine proteases. Ann Biochem 99:53–64

    Article  CAS  Google Scholar 

  7. Cohen AB, Chenoweth E, Hugli TE (1982) The release of elastase, myeloperoxidase, and lysozyme from human alveolar macrophages. Am Rev Respir Dis 126:241–247

    PubMed  CAS  Google Scholar 

  8. Collins JF, de los Santos R, Johanson WG Jr (1986) Acute effects of oleic acid-induced lung injury in baboons. Lung 164:259–268

    Article  PubMed  CAS  Google Scholar 

  9. Crouch TW, Higuchi JH, Coalson JJ, Johanson WG Jr (1984) Pathogenesis and prevention of nosocomial pneumonia in a nonhuman primate model of acute respiratory failure. Am Rev Respir Dis 130:502–504

    PubMed  CAS  Google Scholar 

  10. de los Santos R, Seidenfeld JJ, Coalson JJ, Johanson WG Jr (1985) Antimicrobial prophylaxis against bacterial pneumonia. Am Rev Respir Dis 131:A130

    Google Scholar 

  11. de los Santos R, Seidenfeld JJ, Anzueto A, Collins JF, Coalson JJ, Johanson WG Jr, Peters JI (1987). One hundred percent oxygen lung injury in adult baboons. Am Rev Respir Dis 136:657–661

    PubMed  Google Scholar 

  12. Erlanger BF, Kokowsky N, Cohen W (1961) The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys 95:271–278

    Article  PubMed  CAS  Google Scholar 

  13. Fine R, Collins JF (1982) The effect of theophylline on macrophage elastase secretion. Connect Tissue Res 9:195–199

    Article  PubMed  CAS  Google Scholar 

  14. Fine R, Campbell ME, Collins JF (1984) Elastolytic activity of peripheral blood monocytes. Connect Tissue Res 13:27–35

    PubMed  CAS  Google Scholar 

  15. Fowler AA, Hamman RF, Good JT, Benson KN, Baird M, Eberle D, Petty TL, Hyers TM (1983) The adult respiratory distress syndrome: risk with common predispositions. Ann Intern Med 98:593–597

    PubMed  CAS  Google Scholar 

  16. Galdson M, Levytska V, Liener IE, Twumasi DY (1979) Degradation of tropoelastin and elastin substrates by human neutrophil elastase, free and bound to alpha2-macroglobulin in serum of the M and Z (Pi) phenotypes for alpha1-antitrypsin. Am Rev Respir Dis 119:435–441

    Google Scholar 

  17. Henson PM, Zanolari B, Schwartzman NA, Hong SR (1978) Intracellular control of human neutrophil secretion. I. C5a-induced stimulus-specific desensitization and the effects of cytochalasin B. J Immunol 121:851–855

    PubMed  CAS  Google Scholar 

  18. Idell S, Kucich U, Fein A, Kueppers F, James HL, Walsh PN, Weinbaum G, Coleman RW, Cohen AB (1985) Neutrophil-elastase-releasing factors in bronchoalveolar lavage from patients with adult respiratory distress syndrome. Am Rev Respir Dis 132:1098–1105

    PubMed  CAS  Google Scholar 

  19. Johanson WG Jr, Holcomb JR, Coalson JJ (1982) Experimental diffuse alveolar damage in baboons. Am Rev Respir Dis 126:142–151

    PubMed  Google Scholar 

  20. Johanson WG Jr, Seidenfeld JJ, Gomez P, de los Santos R, Coalson JJ (1988) Bacteriologic diagnosis of nosocomial pneumonia following prolonged mechanical ventilation. Am Rev Respir Dis 137:259–264

    PubMed  Google Scholar 

  21. Johanson WG Jr, Seidenfeld JJ, de los Santos R, Coalson JJ, Gomez P (1988) Prevention of nosocomial pneumonia using topical and parenteral antimicrobial agents. Am Rev Respir Dis 137:265–272

    PubMed  CAS  Google Scholar 

  22. Lee CT, Fein AM, Lippmann M, Holtzman H, Kimbel P, Weinbaum G (1981) Elastolytic activity in pulmonary lavage fluid from patients with adult respiratory distress syndrome. N Engl J Med 304:192–196

    Article  PubMed  CAS  Google Scholar 

  23. Lieberman J (1975) Elevation of serum angiotensin-converting enzyme level in sarcoidosis. Am J Med 59:365–372

    Article  PubMed  CAS  Google Scholar 

  24. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951). Protein measurement with the Folinphenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  25. Matheson NR, Wong PS, Travis J (1979) Enzymatic inactivation of alpha-1-protease inhibitor by neutrophil myeloperoxidase. Biochem Biophys Res Commun 88:402–409

    Article  PubMed  CAS  Google Scholar 

  26. McGuire WM, Spragg RG, Cohen AB, Cochrane CG (1982) Studies on the pathogenesis of the adult respiratory distress syndrome. J Clin Invest 69:543–553

    PubMed  CAS  Google Scholar 

  27. Montgomery AB, Stager MA, Carrico CJ, Hudson LD (1985) Causes of mortality in patients with the adult respiratory distress syndrome. Am Rev Respir Dis 132:485–489

    PubMed  CAS  Google Scholar 

  28. Morihara K, Tsuzuki H, Oka T, Inoue H, Ebata M (1965)Pseudomonas aeruginosa elastase. Isolation, crystallization, and preliminary characterization. J Biol Chem 240:3295–3304

    PubMed  CAS  Google Scholar 

  29. Parsons PE, Fowler AA, Hyers TM, Henson PM (1985) Chemotactic activity in bronchoalveolar lavage fluid from patients with adult respiratory distress syndrome. Am Rev Respir Dis 132:490–493

    PubMed  CAS  Google Scholar 

  30. Revak SD, Rice CL, Schraufstatter IU, Halsey WA Jr, Bohl BP, Clancy RM, Cochrane CG (1985) Experimental pulmonary inflammatory injury in the monkey. J Clin Invest 76:1182–1192

    PubMed  CAS  Google Scholar 

  31. Seidenfeld JJ, Pohl DF, Bell RC, Harris GD, Johanson WG, Jr (1986) Incidence, site, and outcome of infections in ARDS. Am Rev Respir Dis 134:12–16

    PubMed  CAS  Google Scholar 

  32. Senior RM, Campbell EJ, Landis JA, Cox FR, Kuhn C, Koren HS (1982) Elastase of U-937 monocytelike cells. Comparisons with elastases derived from human monocytes and neutrophils and murine macrophagelike cells. J Clin Invest 69:384–393

    Article  PubMed  CAS  Google Scholar 

  33. Travis J (1988) Structure, function, and control of neutrophil proteinases. Am J Med 84(Suppl 6A):37–42

    PubMed  Google Scholar 

  34. Twumasi DY, Liener IE, Galdston M, Levytska V (1977) Activation of human leukocyte elastase by human α2-macroglobulin. Nature 267:61–63

    Article  PubMed  CAS  Google Scholar 

  35. Vissers MCM, George PM, Bathurst IC, Brennan SO, Winterbourn CC (1988) Cleavage and inactivation of α1-antitrypsin by metalloproteinases released from neutrophils. J Clin Invest 82:706–711

    PubMed  CAS  Google Scholar 

  36. Weiland JE, Davis WB, Holter JF, Mohammed JR, Dorinsky PM, Gadek JE (1986) Lung neutrophils in the adult respiratory distress syndrome. Clinical and pathophysiologic significance. Am Rev Respir Dis 133:218–225

    PubMed  CAS  Google Scholar 

  37. Woollen JW, Heyworth R, Walker PG (1961) Studies on glucosaminidase. 3. Testicular N-acetyl-beta-galactosaminidase and N-acetyl-beta-galactosaminidase. Biochem J 78:111–116

    PubMed  CAS  Google Scholar 

  38. Zaslow MC, Clark RA, Stone PJ, Calore JD, Snider GL, Franzblau C (1983) Human neutrophil elastase does not bind to alpha1-protease inhibitor that has been exposed to activated human neutrophils. Am Rev Respir Dis 128:434–439

    PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. J. F. Collins

    Present address: Air Toxics Unit, California Department of Health Services, Berkeley, California, USA

  2. W. G. Johanson

    Present address: University of Texas Medical Branch, Galveston, Texas, USA

Authors and Affiliations

  1. Departments of Medicine (Division of Pulmonary Diseases/Critical Care), Pathology, and Biochemistry, The University of Texas Health Science Center at San Antonio, the Southwest Foundation for Biomedical Research, USA

    J. F. Collins, A. A. Anzueto, J. I. Peters M.D., R. de los Santos, D. C. Gonzalez, W. G. Johanson, J. J. Seidenfeld, J. J. Coalson & S. G. Jenkinson

  2. the Audie L. Murphy Veterans Administration Hospital, 7400 Merton Minter Boulevard, 78284, San Antonio, TX, USA

    J. F. Collins, A. A. Anzueto, J. I. Peters M.D., R. de los Santos, D. C. Gonzalez, W. G. Johanson, J. J. Seidenfeld, J. J. Coalson & S. G. Jenkinson

Authors
  1. J. F. Collins
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. A. Anzueto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. I. Peters M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. de los Santos
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D. C. Gonzalez
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. W. G. Johanson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J. J. Seidenfeld
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. J. J. Coalson
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. S. G. Jenkinson
    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

Collins, J.F., Anzueto, A.A., Peters, J.I. et al. Elastase activity in bronchoalveolar lavage fluid from oxygen-exposed,Pseudomonas-infected baboons. Lung 169, 165–179 (1991). https://doi.org/10.1007/BF02714152

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02714152

Key words

Search

Navigation