title: Neutrophils mediate airway hyperresponsiveness after chlorine-induced airway injury in the mouse.
abstract: Chlorine gas (Cl2) inhalation causes oxidative stress, airway epithelial damage, airway hyperresponsiveness (AHR), and neutrophilia. We evaluated the effect of neutrophil depletion on Cl2-induced AHR and its effect on the endogenous antioxidant response, and if eosinophils or macrophages influence Cl2-induced AHR. We exposed male Balb/C mice to 100 ppm Cl2 for 5 minutes. We quantified inflammatory cell populations in bronchoalveolar lavage (BAL), the antioxidant response in lung tissue by quantitative PCR, and nuclear factor (erythroid-derived 2)-like 2 (NRF2) nuclear translocation by immunofluorescence. In vitro, NRF2 nuclear translocation in response to exogenous hypochlorite was assessed using a luciferase assay. Anti-granulocyte receptor-1 antibody or anti-Ly6G was used to deplete neutrophils. The effects of neutrophil depletion on IL-13 and IL-17 were measured by ELISA. Eosinophils and macrophages were depleted using TRFK5 or clodronate-loaded liposomes, respectively. AHR was evaluated with the constant-phase model in response to inhaled aerosolized methacholine. Our results show that Cl2 exposure induced neutrophilia and increased expression of NRF2 mRNA, superoxide dismutase-1, and heme-oxygenase 1. Neutrophil depletion abolished Cl2-induced AHR in large conducting airways and prevented increases in antioxidant gene expression and NRF2 nuclear translocation. Exogenous hypochlorite administration resulted in increased NRF2 nuclear translocation in vitro. After Cl2 exposure, neutrophils occupied 22 ± 7% of the luminal space in large airways. IL-17 in BAL was increased after Cl2, although this effect was not prevented by neutrophil depletion. Neither depletion of eosinophils nor macrophages prevented Cl2-induced AHR. Our data suggest the ability of neutrophils to promote Cl2-induced AHR is dependent on increases in oxidative stress and occupation of luminal space in large airways.
authors.affiliation: Meakins-Christie Laboratories, Department of Medicine, McGill University, Montreal, Quebec, Canada.
authors.forename: Toby K
authors.initials: TK
authors.lastname: McGovern
authors.forename: Madison
authors.initials: M
authors.lastname: Goldberger
authors.forename: Benoit
authors.initials: B
authors.lastname: Allard
authors.forename: Soroor
authors.initials: S
authors.lastname: Farahnak
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Hamamoto
authors.forename: Michael
authors.initials: M
authors.lastname: O'Sullivan
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Hirota
authors.forename: Guy
authors.initials: G
authors.lastname: Martel
authors.forename: Simon
authors.initials: S
authors.lastname: Rousseau
authors.forename: James G
authors.initials: JG
authors.lastname: Martin
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D059366:Asthma, Occupational / Q000139:chemically induced / Q000276:immunology*; D002713:Chlorine / Q000633:toxicity*; D004002:Clodronic Acid / Q000008:administration & dosage; D016207:Cytokines / Q000378:metabolism; D006098:Granulocytes / Q000276:immunology; D008168:Lung / Q000276:immunology / Q000473:pathology; D008297:Male; D008807:Mice, Inbred BALB C; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology*; D018384:Oxidative Stress; D015854:Up-Regulation
chemical_list: D016207:Cytokines; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D004002:Clodronic Acid; D002713:Chlorine
keywords: anti–granulocyte receptor-1; clodronate-liposomes; eosinophils; nuclear factor (erythroid-derived 2)-like 2; oxidative stress