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Original Research |

Toll-like receptor 3 stimulation causes corticosteroid-refractory airway neutrophilia and hyperresponsiveness in mice

Genki Kimura, MSc; Keitaro Ueda; Shouichi Eto; Yuji Watanabe; Takashi Masuko, PhD; Tadashi Kusama; Peter J. Barnes, MD, FCCP; Kazuhiro Ito, DVM, PhD; Yasuo Kizawa
Author and Funding Information

Department of Physiology and Anatomy, Nihon University School of Pharmacy, Funabashi, Chiba 274-8555, Japan (Kimura, Ueda, Eto, Watanabe, Masuko, Kusama, Kizawa); Airway Disease Section, National Heart and Lung Institute, Imperial College, Dovehouse Street, London SW3 6LY, United Kingdom (Barnes, Ito)

Corresponding author: Yasuo Kizawa, PhD, Department of Physiology and Anatomy, Nihon University School of Pharmacy, 7-7-1, Narashinodai, Funabashi, Chiba 274-8555, Japan; e-mail address: kizawa.yasuo@nihon-u.ac.jp

Sources of funding: This research was funded by “High-Tech Research Center” Project for Private Universities (2007-2011): matching fund subsidy from MEXT (Japan; to Dr Kizawa).


Chest. 2013. doi:10.1378/chest.12-2610
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Abstract

Background:  RNA virus infections, such as rhinovirus and respiratory syncytial virus, induce exacerbations in patients with chronic obstructive pulmonary disease (COPD) and asthma, and the inflammation is corticosteroid refractory. The main aim of this study is to establish a murine model induced by a Toll-like receptor 3 (TLR3) agonist, a RNA virus mimic, and investigate the response to corticosteroid.

Methods:  A/J mice were given polyinosinic-polycytidylic acid (poly(I:C)), a TLR3 agonist, intranasally, in the presence or absence of cigarette smoke exposure. Inflammatory cell accumulation and CXCL1, IFNs and CXCL10 productions in bronchoalveolar lavage fluid (BALF) were determined by flow cytometry and ELISA, respectively, and airway hyperresponsiveness (AHR) to histamine/methacholine was determined by a two-chambered, double-flow plethysmography system. BALB/c and C57BL/6J mice were also used for comparisons.

Results:  Intranasal treatment of poly(I:C) significantly induced airway neutrophilia, production of CXCL1, IFN-β and CXCL10, necrotic cell accumulation in BALF, and also increased airway responsiveness to histamine or methacholine inhalation. These poly(I:C)-dependent airway inflammation and AHR were not inhibited by the corticosteroid fluticasone propionate (FP; up to 0.5 mg/ml intranasal), although FP strongly inhibited LPS (TLR4 agonist)-induced airway neutrophilia. Furthermore, cigarette smoke exposure significantly increased TLR3 expression in murine lung tissue, and exacerbated poly(I:C)-induced neutrophilia and AHR.

Conclusions:  These results suggest that TLR3 stimulation is involved in corticosteroid refractory airway inflammation in lung, which is enhanced by cigarette smoking, and this may provide a model for understanding virus induced exacerbations in COPD and their therapy.


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