Chest
Volume 107, Issue 6, June 1995, Pages 1641-1647
Journal home page for Chest

Clinical Investigations: Lavage
Human Neutrophil Collagenase (MMP-8), Identified in Bronchiectasis BAL Fluid, Correlates With Severity of Disease

https://doi.org/10.1378/chest.107.6.1641Get rights and content

Collagenases in bronchoalveolar lavage fluid (BALF) of patients with bronchiectasis and healthy subjects were characterized using specific functional and immunologic assays. The BAL fluid contained interstitial collagenase and collagenolytic proteinases of bacterial origin. Collagenase activities, obtained after organomercurial activation, correlated with the severity of bronchiectasis. In severe cases, collagenase activities were 3.5×10−7 IU/L/48 h or 4.8×10−6 IU/g/48 h (p<0.01), in moderate ones 1.74×10−7 IU/L/48 h or 3.35×10−6 IU/g/48 h (p<0.05), and in mild cases 0.32×10−7 IU/L/48 h or 0.7×10−6 IU/g/48 h (p<0.05). The corresponding activities in healthy control subjects were 0.08×10−7 IU/L/48 h or 0.13×10−6 IU/g/48 h. The cellular origin of interstitial collagenase was assessed with doxycycline inhibition test utilizing the differential sensitivity of fibroblast-type collagenase/MMP-1 (IC50=280 μM) and neutrophil-type collagenase/MMP-8 (IC50=26 μM) to the anticollagenolytic, nonantimicrobial doxycycline action. Interstitial collagenase, contained in BALF, was totally inhibited by 100 μM of doxycycline. It can therefore be concluded that most of mammalian collagenase presented in inflamed fluid of bronchiectasis originated from neutrophils. The molecular forms of neutrophil-type collagenase/MMP-8 were confirmed and analyzed by Western-blot, which showed evidence of the proteolytic conversion of the latent 85-kD MMP-8 proenzyme species into active 65-kD molecular weight species. These findings strongly suggest involvement of proteolytic activation pathway of proMMP-8, especially in severe and moderate forms of bronchiectasis. Furthermore, collagenolytic proteases of bacterial origins may also participate in tissue destruction of the lung.

Section snippets

Methods

Thirty-six patients with bronchiectasis referred to the Department of Lung Diseases of Tartu University (Estonia) were included in this study. The clinical diagnosis was based on the prolonged history of frequent bronchitis and/or pneumonia and intermittent sputum production, the persistence of crackles on auscultation, and cystic changes on CT and/or bronchograms.

Patients were divided into three different disease activity subgroups based on the clinical characteristics and lung CT findings:

Functional Activities of Collagenases in BALF of Patients With Bronchiectasis and Healthy Control Subjects

Collagenolytic activities, obtained after optimal organomercurial (PMC) activation (Fig 1) and autoactive (Fig 2) collagenolytic activities were determined after 48 h incubation of the fluid obtained by BAL (nBE=36 and nHC=l5), and 1.2 μM type I collagen with and without 1 mM PMC, respectively. Collagenolytic activities, obtained after organomercurial activation, differed significantly in different BE subgroups. The corresponding data were the following: in mild cases, 0.32×10−7 IU/L/48 h or

Discussion

Development of bronchiectases during the ongoing lung inflammation is considered to be associated with an excess of catalytically active proteinases1, 2, 3, 4, 5 in target tissues. Circulating inflammatory cells, polymorphonuclear leukocytes (PMNs), and monocytes,21, 45 as well as pulmonary resident cells, fibroblasts, epithelial cells, and alveolar macrophages, synthesize and release various proteinases.17, 19, 20 Activation of such locally released proteinases exerts the potential to result

References (53)

  • HillSL et al.

    The response of patients with purulent bronchiectasis to antibiotics for 4 months

    Q J Med

    (1988)
  • TetleyTD

    New perspectives on basic mechanisms in lung disease: VI

    Proteinase imbalance: its role in lung disease. Thorax

    (1993)
  • LuceyEC et al.

    Consequences of proteolytic injury

  • CrystalRG et al.

    Structural organization of the pulmonary interstitium

  • BienkowskiRS

    Interstitial collagens

  • MayneR et al.

    Structure and function of collagen types

    (1987)
  • WoessnerJFJ

    Matrix metalloproteinases and their inhibitors in connective tissue remodeling

    FASEB J

    (1991)
  • Birkedal-HansenH et al.

    Matrix metalloproteinases: a review

    Clin Rev Oral Biol Med

    (1993)
  • MurphyG et al.

    The matrix metalloproteinases and their inhibitors

    Am J Respir Cell Mol Biol

    (1992)
  • MillerEJ et al.

    Cleavage of type II and III collagens with mammalian collagenase: site of cleavage and primary structure at the NH2-terminal portion of the smaller fragment released from both collagens

    Biochemistry

    (1976)
  • StricklinGP et al.

    Human skin collagenase: isolation of precursor and active forms from both fibroblast and organ cultures

    Biochemistry

    (1977)
  • ShapiroSD et al.

    Proteinases secreted by human mononuclear phagocytes

    J Rheumatol

    (1991)
  • WelgusHG et al.

    Neutral metalloproteinases produced by human mononuclear phagocytes: enzyme profile, regulation, and expression during cellular development

    J Clin Invest

    (1990)
  • WelgusHG et al.

    Human alveolar macrophages produce a fibroblast-like collagenase and collagenase inhibitor

    J Clin Invest

    (1985)
  • HastyKA et al.

    Heterogeneity among human collagenases demonstrated by monoclonal antibody that selectively recognizes and inhibits human neutrophil collagenase

    J Exp Med

    (1984)
  • GolubLM et al.

    Host modulation with tetracyclines and their chemically modified analogues

    Curr Opin Dent

    (1992)
  • Cited by (92)

    • Future Directions in Bronchiectasis Research

      2022, Clinics in Chest Medicine
      Citation Excerpt :

      Several studies have reported elevated levels of MMP-8 and MMP-9, produced by neutrophils, in the airways of patients with bronchiectasis. Their increase has been associated with the inflammatory process and the epithelial destruction characteristic of these diseases.25–27 With regard to the symptoms, elevated levels of MMP-8 and MMP-9 have been associated with a greater radiological extension and a greater bacterial load in sputum, worse FEV1, greater severity of the disease, increased sputum purulence, presence of airway P. aeruginosa infection and prediction of future exacerbations.28

    • Bronchiectasis and Chronic Suppurative Lung Disease

      2019, Kendig's Disorders of the Respiratory Tract in Children
    • Human matrix metalloproteinases: An ubiquitarian class of enzymes involved in several pathological processes

      2012, Molecular Aspects of Medicine
      Citation Excerpt :

      Whereas MMP-8 levels are significantly increased in BALF from patients with subclinical emphysema (Betsuyaku et al., 1999), levels of MMP-8 in the sputum of patients with emphysema correlate negatively with lung function if the patients have developed chronic obstructive pulmonary disease (COPD) (Culpitt et al., 2005). Finally, MMP-8 levels in BALF also correlate with the severity of bronchiectasis, a disease characterized by a permanent dilation of the bronchi due to chronic inflammation, leading to irreparable deformation of bronchial wall components (Sepper et al., 1995). Other studies identified activated epithelial cells (as well as infiltrating neutrophils and macrophages) as the cell types responsible for the de novo MMP-8 expression in the course of this pathology (Prikk et al., 2001; Zheng et al., 2002).

    • Bronchiectasis and Chronic Suppurative Lung Disease

      2012, Kendig and Chernick's Disorders of the Respiratory Tract in Children
    • Bronchiectasis and sino-nasal disease: A review

      2008, Journal of Laryngology and Otology
    View all citing articles on Scopus

    Supported by Academy of Finland and Ministry of Social and Health Affairs of Finland.

    revision accepted October 18.

    View full text