Chest
Volume 107, Issue 6, June 1995, Pages 1641-1647
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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.

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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

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

    revision accepted October 18.

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