Am. J. Respir. Cell Mol. Biol., Volume 20, Number 4, April, 1999 729-736

Proteinase 3, a Potent Secretagogue in Airways, Is Present in Cystic Fibrosis Sputum

Véronique Witko-Sarsat, Lise Halbwachs-Mecarelli, Antje Schuster, Patrick Nusbaum, Iris Ueki, Sandrine Canteloup, Gérard Lenoir, Béatrice Descamps-Latscha, and Jay A. Nadel

INSERM U 90, Hôpital des Enfants Malades, Paris, France; Children's Hospital, University of Düsseldorf, Germany; and Cardiovascular Research Institute and Department of Medicine and Physiology, University of California, San Francisco

We evaluated the roles of proteinase 3 (PR3) and human neutrophil elastase (HNE), two neutrophil serine proteinases in the mechanisms leading to airway inflammation and hypersecretion in cystic fibrosis (CF). Using specific enzyme-linked immunosorbent assay (ELISA), we found higher levels of PR3 than HNE in sputum from CF patients. Using two inhibitors, ICI (Imperial Chemical Industries) 200,355 (which inhibits both HNE and PR3) and secretory leukoproteinase inhibitor (SLPI) (which inhibits only HNE), we showed that PR3 was enzymatically active in sputum, and its activity, as assessed by SLPI-resistant serine proteinase activity, correlated highly with its antigenic concentration measured by ELISA. Interestingly, sputum pellet-associated serine proteinase activity was mostly due to HNE. PR3 purified from neutrophil azurophil granules triggered airway gland secretion, as measured by the release of radiolabeled molecules from cultured bovine tracheal serous cells pulse-labeled with Na₂³⁵SO₄. This secretory activity was inhibited by ICI 200,355. PR3 concentration in CF sputum was highly correlated with taurine concentration, a reliable marker of airway inflammation and respiratory scores (e.g., FEV₁%), whereas no significant correlation was observed with HNE. We verified that Pseudomonas aeruginosa proteinases did not interfere with the assessment of PR3 and HNE. Indeed, the PR3/HNE ratio was greatest in patients chronically infected by P. aeruginosa. We suggest that PR3 may play a role in the hypersecretory process that is characteristic of CF.

This article has been cited by other articles:

				J. Cooley, B. McDonald, F. J. Accurso, E. C. Crouch, and E. Remold-O'Donnell Patterns of neutrophil serine protease-dependent cleavage of surfactant protein D in inflammatory lung disease J. Leukoc. Biol., April 1, 2008; 83(4): 946 - 955. [Abstract] [Full Text] [PDF]

				S. Attucci, A. Gauthier, B. Korkmaz, P. Delepine, M. F.-D. Martino, F. Saudubray, P. Diot, and F. Gauthier EPI-hNE4, a Proteolysis-Resistant Inhibitor of Human Neutrophil Elastase and Potential Anti-Inflammatory Drug for Treating Cystic Fibrosis J. Pharmacol. Exp. Ther., August 1, 2006; 318(2): 803 - 809. [Abstract] [Full Text] [PDF]

				B. Dublet, A. Ruello, M. Pederzoli, E. Hajjar, M. Courbebaisse, S. Canteloup, N. Reuter, and V. Witko-Sarsat Cleavage of p21/WAF1/CIP1 by Proteinase 3 Modulates Differentiation of a Monocytic Cell Line: MOLECULAR ANALYSIS OF THE CLEAVAGE SITE J. Biol. Chem., August 26, 2005; 280(34): 30242 - 30253. [Abstract] [Full Text] [PDF]

				M. Pederzoli, C. Kantari, V. Gausson, S. Moriceau, and V. Witko-Sarsat Proteinase-3 Induces Procaspase-3 Activation in the Absence of Apoptosis: Potential Role of this Compartmentalized Activation of Membrane-Associated Procaspase-3 in Neutrophils J. Immunol., May 15, 2005; 174(10): 6381 - 6390. [Abstract] [Full Text] [PDF]

				P. J. Barnes Mediators of Chronic Obstructive Pulmonary Disease Pharmacol. Rev., December 1, 2004; 56(4): 515 - 548. [Abstract] [Full Text] [PDF]

				B. Korkmaz, S. Attucci, T. Moreau, E. Godat, L. Juliano, and F. Gauthier Design and Use of Highly Specific Substrates of Neutrophil Elastase and Proteinase 3 Am. J. Respir. Cell Mol. Biol., June 1, 2004; 30(6): 801 - 807. [Abstract] [Full Text] [PDF]

				F Rubio, J Cooley, F J Accurso, and E Remold-O'Donnell Linkage of neutrophil serine proteases and decreased surfactant protein-A (SP-A) levels in inflammatory lung disease Thorax, April 1, 2004; 59(4): 318 - 323. [Abstract] [Full Text] [PDF]

				S. Durant, M. Pederzoli, Y. Lepelletier, S. Canteloup, P. Nusbaum, P. Lesavre, and V. Witko-Sarsat Apoptosis-induced proteinase 3 membrane expression is independent from degranulation J. Leukoc. Biol., January 1, 2004; 75(1): 87 - 98. [Abstract] [Full Text] [PDF]

				P.J. Barnes, S.D. Shapiro, and R.A. Pauwels Chronic obstructive pulmonary disease: molecular and cellularmechanisms Eur. Respir. J., October 1, 2003; 22(4): 672 - 688. [Abstract] [Full Text] [PDF]

				H. Corvol, C. Fitting, K. Chadelat, J. Jacquot, O. Tabary, M. Boule, J.-M. Cavaillon, and A. Clement Distinct cytokine production by lung and blood neutrophils from children with cystic fibrosis Am J Physiol Lung Cell Mol Physiol, June 1, 2003; 284(6): L997 - L1003. [Abstract] [Full Text] [PDF]

				V. Witko-Sarsat, S. Canteloup, S. Durant, C. Desdouets, R. Chabernaud, P. Lemarchand, and B. Descamps-Latscha Cleavage of p21waf1 by Proteinase-3, a Myeloid-specific Serine Protease, Potentiates Cell Proliferation J. Biol. Chem., November 27, 2002; 277(49): 47338 - 47347. [Abstract] [Full Text] [PDF]

				B. Korkmaz, S. Attucci, E. Hazouard, M. Ferrandiere, M. L. Jourdan, M. Brillard-Bourdet, L. Juliano, and F. Gauthier Discriminating between the Activities of Human Neutrophil Elastase and Proteinase 3 Using Serpin-derived Fluorogenic Substrates J. Biol. Chem., October 11, 2002; 277(42): 39074 - 39081. [Abstract] [Full Text] [PDF]

				O. E. Sorensen, P. Follin, A. H. Johnsen, J. Calafat, G. S. Tjabringa, P. S. Hiemstra, and N. Borregaard Human cathelicidin, hCAP-18, is processed to the antimicrobial peptide LL-37 by extracellular cleavage with proteinase 3 Blood, June 15, 2001; 97(12): 3951 - 3959. [Abstract] [Full Text] [PDF]

				Y. M. van der Geld, P. C. Limburg, and C. G. M. Kallenberg Proteinase 3, Wegener's autoantigen: from gene to antigen J. Leukoc. Biol., February 1, 2001; 69(2): 177 - 190. [Abstract] [Full Text]

				Q.-L. Ying and S. R. Simon Kinetics of the Inhibition of Proteinase 3 by Elafin Am. J. Respir. Cell Mol. Biol., January 1, 2001; 24(1): 83 - 89. [Abstract] [Full Text]

				A. Hill, S. Gompertz, and R. Stockley Factors influencing airway inflammation in chronic obstructive pulmonary disease Thorax, November 1, 2000; 55(11): 970 - 977. [Full Text]

				E. J. Campbell, M. A. Campbell, and C. A. Owen Bioactive Proteinase 3 on the Cell Surface of Human Neutrophils: Quantification, Catalytic Activity, and Susceptibility to Inhibition J. Immunol., September 15, 2000; 165(6): 3366 - 3374. [Abstract] [Full Text] [PDF]

				P. J. Barnes Chronic Obstructive Pulmonary Disease N. Engl. J. Med., July 27, 2000; 343(4): 269 - 280. [Full Text] [PDF]

				V. Witko-Sarsat, E. M. Cramer, C. Hieblot, J. Guichard, P. Nusbaum, S. Lopez, P. Lesavre, and L. Halbwachs-Mecarelli Presence of Proteinase 3 in Secretory Vesicles: Evidence of a Novel, Highly Mobilizable Intracellular Pool Distinct From Azurophil Granules Blood, October 1, 1999; 94(7): 2487 - 2496. [Abstract] [Full Text] [PDF]