Evaluation of active neutrophil elastase in sputum of bronchiectasis and cystic fibrosis patients: A comparison among different techniques

Martina Oriano, Leonardo Terranova, Giovanni Sotgiu, Laura Saderi, Angela Bellofiore, Mariangela Retucci, Cinzia Marotta, Andrea Gramegna, Daniela Miglietta, Chiara Carnini, Paola Marchisio, James D. Chalmers, Stefano Aliberti, Francesco Blasi

Research output: Contribution to journalArticle

Abstract

Neutrophil elastase (NE) is a crucial marker of neutrophilic inflammation. We aimed to compare different techniques to detect active NE in sputum samples of 50 Bronchiectasis (BE) and 50 Cystic Fibrosis (CF) patients. Three methods including a ProteaseTag® Active NE Immunoassay (ELISA) and two enzymatic digestion assays (chromogenic –CS- and fluorogenic –FS- substrate) were compared. Results of active NE were also correlated with clinical data. The three methods provided statistically different values for NE activity in the same sputum samples in both cohorts. In the BE cohort, the highest correlations between NE activity and Bronchiectasis Severity Index (rho = 0.40, P < 0.0001), sputum purulence (AUC = 0.79), and chronic infections due to any pathogen (AUC = 0.76) and P. aeruginosa (AUC = 0.80) were found when NE was measured through the activity-based immunoassay. In the CF cohort, the highest correlations between NE activity and sputum quantity (rho = 0.71) and FEV1% (rho = 0.42, P = 0.03) were observed when the FS method was used, while similar correlations with chronic P. aeruginosa infection were identified with the FS and ELISA methods. NE activity in sputum correlates with clinical variables in both diseases. However, different methods to evaluate active NE in sputum lead to significantly different results, also in terms of correlation with clinical data.

Original languageEnglish
Article number101856
JournalPulmonary Pharmacology and Therapeutics
Volume59
DOIs
Publication statusPublished - Dec 1 2019

Fingerprint

Leukocyte Elastase
Bronchiectasis
Sputum
Cystic Fibrosis
Area Under Curve
Immunoassay
Enzyme-Linked Immunosorbent Assay
Chromogenics
Enzyme Assays
Pathogens
Infection
Fluorescent Dyes
Digestion
Assays
Inflammation

Keywords

  • Bronchiectasis
  • Cystic fibrosis
  • Lung inflammation
  • Neutrophil elastase

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Biochemistry, medical
  • Pharmacology (medical)

Cite this

Evaluation of active neutrophil elastase in sputum of bronchiectasis and cystic fibrosis patients : A comparison among different techniques. / Oriano, Martina; Terranova, Leonardo; Sotgiu, Giovanni; Saderi, Laura; Bellofiore, Angela; Retucci, Mariangela; Marotta, Cinzia; Gramegna, Andrea; Miglietta, Daniela; Carnini, Chiara; Marchisio, Paola; Chalmers, James D.; Aliberti, Stefano; Blasi, Francesco.

In: Pulmonary Pharmacology and Therapeutics, Vol. 59, 101856, 01.12.2019.

Research output: Contribution to journalArticle

Oriano, Martina ; Terranova, Leonardo ; Sotgiu, Giovanni ; Saderi, Laura ; Bellofiore, Angela ; Retucci, Mariangela ; Marotta, Cinzia ; Gramegna, Andrea ; Miglietta, Daniela ; Carnini, Chiara ; Marchisio, Paola ; Chalmers, James D. ; Aliberti, Stefano ; Blasi, Francesco. / Evaluation of active neutrophil elastase in sputum of bronchiectasis and cystic fibrosis patients : A comparison among different techniques. In: Pulmonary Pharmacology and Therapeutics. 2019 ; Vol. 59.
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abstract = "Neutrophil elastase (NE) is a crucial marker of neutrophilic inflammation. We aimed to compare different techniques to detect active NE in sputum samples of 50 Bronchiectasis (BE) and 50 Cystic Fibrosis (CF) patients. Three methods including a ProteaseTag{\circledR} Active NE Immunoassay (ELISA) and two enzymatic digestion assays (chromogenic –CS- and fluorogenic –FS- substrate) were compared. Results of active NE were also correlated with clinical data. The three methods provided statistically different values for NE activity in the same sputum samples in both cohorts. In the BE cohort, the highest correlations between NE activity and Bronchiectasis Severity Index (rho = 0.40, P < 0.0001), sputum purulence (AUC = 0.79), and chronic infections due to any pathogen (AUC = 0.76) and P. aeruginosa (AUC = 0.80) were found when NE was measured through the activity-based immunoassay. In the CF cohort, the highest correlations between NE activity and sputum quantity (rho = 0.71) and FEV1{\%} (rho = 0.42, P = 0.03) were observed when the FS method was used, while similar correlations with chronic P. aeruginosa infection were identified with the FS and ELISA methods. NE activity in sputum correlates with clinical variables in both diseases. However, different methods to evaluate active NE in sputum lead to significantly different results, also in terms of correlation with clinical data.",
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AU - Terranova, Leonardo

AU - Sotgiu, Giovanni

AU - Saderi, Laura

AU - Bellofiore, Angela

AU - Retucci, Mariangela

AU - Marotta, Cinzia

AU - Gramegna, Andrea

AU - Miglietta, Daniela

AU - Carnini, Chiara

AU - Marchisio, Paola

AU - Chalmers, James D.

AU - Aliberti, Stefano

AU - Blasi, Francesco

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AB - Neutrophil elastase (NE) is a crucial marker of neutrophilic inflammation. We aimed to compare different techniques to detect active NE in sputum samples of 50 Bronchiectasis (BE) and 50 Cystic Fibrosis (CF) patients. Three methods including a ProteaseTag® Active NE Immunoassay (ELISA) and two enzymatic digestion assays (chromogenic –CS- and fluorogenic –FS- substrate) were compared. Results of active NE were also correlated with clinical data. The three methods provided statistically different values for NE activity in the same sputum samples in both cohorts. In the BE cohort, the highest correlations between NE activity and Bronchiectasis Severity Index (rho = 0.40, P < 0.0001), sputum purulence (AUC = 0.79), and chronic infections due to any pathogen (AUC = 0.76) and P. aeruginosa (AUC = 0.80) were found when NE was measured through the activity-based immunoassay. In the CF cohort, the highest correlations between NE activity and sputum quantity (rho = 0.71) and FEV1% (rho = 0.42, P = 0.03) were observed when the FS method was used, while similar correlations with chronic P. aeruginosa infection were identified with the FS and ELISA methods. NE activity in sputum correlates with clinical variables in both diseases. However, different methods to evaluate active NE in sputum lead to significantly different results, also in terms of correlation with clinical data.

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