title: Alpha-1 Antitrypsin Mitigates the Inhibition of Airway Epithelial Cell Repair by Neutrophil Elastase.
abstract: Neutrophil elastase (NE) activity is associated with many destructive lung diseases and is a predictor for structural lung damage in early cystic fibrosis (CF), which suggests normal maintenance of airway epithelium is prevented by uninhibited NE. However, limited data exist on how the NE activity in airways of very young children with CF affects function of the epithelia. The aim of this study was to determine if NE activity could inhibit epithelial homeostasis and repair and whether any functional effect was reversible by antiprotease alpha-1 antitrypsin (α1AT) treatment. Viability, inflammation, apoptosis, and proliferation were assessed in healthy non-CF and CF pediatric primary airway epithelial cells (pAECnon-CF and pAECCF, respectively) during exposure to physiologically relevant NE. The effect of NE activity on pAECCF wound repair was also assessed. We report that viability after 48 hours was significantly decreased by 100 nM NE in pAECnon-CF and pAECCF owing to rapid cellular detachment that was accompanied by inflammatory cytokine release. Furthermore, both phenotypes initiated an apoptotic response to 100 nM NE, whereas ≥ 50 nM NE activity significantly inhibited the proliferative capacity of cultures. Similar concentrations of NE also significantly inhibited wound repair of pAECCF, but this effect was reversed by the addition of α1AT. Collectively, our results demonstrate free NE activity is deleterious for epithelial homeostasis and support the hypothesis that proteases in the airway contribute directly to CF structural lung disease. Our results also highlight the need to investigate antiprotease therapies in early CF disease in more detail.
authors.affiliation: 1 School of Paediatrics and Child Health.|2 Telethon Kids Institute.
authors.forename: Luke W
authors.initials: LW
authors.lastname: Garratt
authors.affiliation: 2 Telethon Kids Institute.|3 Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia, Australia.
authors.forename: Erika N
authors.initials: EN
authors.lastname: Sutanto
authors.affiliation: 2 Telethon Kids Institute.
authors.forename: Kak-Ming
authors.initials: KM
authors.lastname: Ling
authors.affiliation: 1 School of Paediatrics and Child Health.
authors.forename: Kevin
authors.initials: K
authors.lastname: Looi
authors.affiliation: 1 School of Paediatrics and Child Health.|4 Centre for Cell Therapy and Regenerative Medicine, and.
authors.forename: Thomas
authors.initials: T
authors.lastname: Iosifidis
authors.affiliation: 2 Telethon Kids Institute.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Martinovich
authors.affiliation: 2 Telethon Kids Institute.
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Shaw
authors.affiliation: 2 Telethon Kids Institute.|5 Centre for Microscopy, Characterisation and Analysis, University of Western Australia, Nedlands, Perth, Western Australia, Australia.
authors.forename: Alysia G
authors.initials: AG
authors.lastname: Buckley
authors.affiliation: 2 Telethon Kids Institute.|3 Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia, Australia.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Kicic-Starcevich
authors.affiliation: 1 School of Paediatrics and Child Health.|6 School of Medicine, Notre Dame University, Fremantle, Perth, Western Australia, Australia.
authors.forename: Francis J
authors.initials: FJ
authors.lastname: Lannigan
authors.affiliation: 7 School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, New South Wales, Australia.|8 Priority Research Centre for Asthma and Respiratory Disease, Hunter Medical Research Institute, Newcastle, New South Wales, Australia.|9 Department of Anesthesiology, Pharmacology and Therapeutics, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Darryl A
authors.initials: DA
authors.lastname: Knight
authors.affiliation: 1 School of Paediatrics and Child Health.|2 Telethon Kids Institute.|4 Centre for Cell Therapy and Regenerative Medicine, and.|3 Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia, Australia.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Stick
authors.affiliation: 1 School of Paediatrics and Child Health.|2 Telethon Kids Institute.|4 Centre for Cell Therapy and Regenerative Medicine, and.|3 Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia, Australia.|10 Department of Respiratory Medicine, Royal Children's Hospital, Parkville, Melbourne, Victoria, Australia; and.|11 Murdoch Childrens Research Institute, Parkville, Melbourne, Victoria, Australia.
authors.forename: Anthony
authors.initials: A
authors.lastname: Kicic
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D016022:Case-Control Studies; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000201:enzymology* / Q000473:pathology; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D018836:Inflammation Mediators / Q000378:metabolism; D019272:Leukocyte Elastase / Q000494:pharmacology*; D008297:Male; D010641:Phenotype; D012038:Regeneration / Q000187:drug effects*; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D013997:Time Factors; D000515:alpha 1-Antitrypsin / Q000494:pharmacology*
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators; C043392:SERPINA1 protein, human; D000515:alpha 1-Antitrypsin; D019272:Leukocyte Elastase
keywords: alpha-1 antitrypsin; cystic fibrosis; neutrophil elastase; primary cell culture; wound repair