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Am J Physiol Lung Cell Mol Physiol 276: L28-L34, 1999;
1040-0605/99 $5.00
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Vol. 276, Issue 1, L28-L34, January 1999

Neutrophil elastase and elastase-rich cystic fibrosis sputum degranulate human eosinophils in vitro

Hong Liu, Stephen C. Lazarus, George H. Caughey, and John V. Fahy

Department of Medicine and Cardiovascular Research Institute, University of California, San Francisco, California 94143

Neutrophils, eosinophils, and their proinflammatory constituents are important mediators of airway disease, and high levels of neutrophil proteases and eosinophil cationic protein (ECP) are found in sputum from patients with cystic fibrosis (CF). To investigate whether neutrophil proteases or CF sputum causes eosinophil degranulation, purified eosinophils from atopic asthmatic subjects were incubated for 2 h with neutrophil elastase, cathepsin G, and CF sputum, and the release of ECP was measured. We found that the percent release of ECP was higher after incubation with neutrophil elastase (10-5 M) than with a buffer control [6.1 ± 0.8 (SE) vs. 1.7 ± 0.1%; P < 0.003] and represented >50% of the release caused by positive controls [Ca2+ ionophore A-23187 (5 × 10-6 M) or serum-coated Sephadex beads]. The release of ECP after incubation with cathepsin G (2.3 ± 0.2%) and CF sputum (6.2 ± 2.0%) was also significantly higher than that with a buffer control (P < 0.05). Neutralization of free elastase activity with alpha 1-proteinase inhibitor reduced the mean percent degranulation of eosinophils by neutrophil elastase by 50% (P = 0.0004) and by CF sputum by 75% (P = 0.02). Preincubation of eosinophils with cytochalasin B (10 mg/ml) and depletion of the incubation medium of Ca2+ also significantly attenuated degranulation of eosinophils incubated with purified free neutrophil elastase or CF sputum (P < 0.05). We conclude that neutrophil proteases, especially neutrophil elastase, and elastase-rich CF sputum cause degranulation of eosinophils in a mechanism partially dependent on Ca2+ and actin filaments.

eosinophil cationic protein


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