TY - JOUR T1 - Alpha-1 antitrypsin ameliorates inhibition of wound repair by neutrophil elastase JF - European Respiratory Journal JO - Eur Respir J VL - 44 IS - Suppl 58 SP - P3466 AU - Luke Garratt AU - Erika Sutanto AU - Kak-Ming Ling AU - Kevin Looi AU - Elizabeth Kicic-Starcevich AU - Thomas Iosifidis AU - Kelly Martinovich AU - Francis Lannigan AU - Stephen Stick AU - Anthony Kicic Y1 - 2014/09/01 UR - http://erj.ersjournals.com/content/44/Suppl_58/P3466.abstract N2 - Introduction: Cystic fibrosis (CF) is characterised by increasing free active neutrophil elastase (NE) that is significantly associated with bronchiectasis. Yet data on how NE affects function of primary epithelia in very young children is limited. Aim: Do physiologically relevant, nanomolar (nM) concentrations of NE inhibit repair of airway epithelial cells (AEC) in a manner reversible by the antiprotease alpha 1-antitrypsin (A1AT). Method: Non-CF (NuLi-1 & pAECNCF) and CF (CuFi-1 & pAECCF) AECs were used, with primary pAECNCF and pAECCF collected from paediatric airways (mean age 4.67 and 4.22 years respectively). Cells were exposed to 10-200 nM NE over 48-hours and viability, proliferative capacity, cytotoxicity and wound repair were assessed. Results: Following exposure to 100 nM NE, pAECNCF and pAECCF viability (n = 4; 3.9% ±5.1, p<0.01 and 40.0% ±30.4, p<0.05) and proliferation (n = 4; 16.5% ±5.4, p<0.01 and 13.2% ±0.2, p<0.01) were significantly reduced compared to controls. Furthermore, 200 nM NE inhibited NuLi-1 and CuFi-1 scratch wound closure at 48 hours (n = 6; 32.01% of control, p<0.01 and 17.75% of control, p<0.01) and 50 nM NE inhibited pAECCF repair at 24 hours (n = 2; 69.99% of control, p<0.01). Addition of 1 µM A1AT at 12-hours post-wounding significantly reduced inhibition by NE at all concentrations assessed. Conclusion: Biologically relevant concentrations of NE are associated with cytotoxicity, reduced proliferation and deficient wound repair providing a mechanism for bronchiectasis formation. Current anti-inflammation therapies may not be sufficient to reduce the impact of excess proteases in the CF airway and targeting protease activity offers an attractive therapeutic option. ER -