Download European Respiratory Society Annual Congress 2013

European Respiratory Society
Annual Congress 2013
Abstract Number: 2862
Publication Number: P3128
Abstract Group: 3.2. Airway Cell Biology and Immunopathology
Keyword 1: Inflammation Keyword 2: Pharmacology Keyword 3: Smoking
Title: Anti-inflammatory effects of roflumilast N-oxide and dexamethasone in human bronchial epithelial
cells stimulated with toll-like receptor agonists
Javier 12009 Milara [email protected] 1, Javier 12022 Lluch [email protected] 1, Patricia
12023 Almudever [email protected] 1, Teresa 12024 Peiró [email protected] 1, Adela 12025
Serrano [email protected] 1, Magdalena 12087 Alonso-Galicia
[email protected] 2, Jose 12091 Freire [email protected] 2, Xiaozhong 12092 Qian
[email protected] 2 and Julio 12093 Cortijo [email protected] 1. 1 Pharmacology, U Valencia, Fundación
De Investigación Del Hospital General De Valencia, Valencia, Spain and 2 Pharmacology and Discovery,
Forest Research Institute, Jersey City, United States .
Body: Objective Bacterial and viral infections in COPD contribute to inflammation and exacerbations, and
toll-like receptors (TLR) modulate the innate immune response to these infectious agents. This study
explored the anti-inflammatory effects of dexamethasone (DEX) and roflumilast N-oxide (RNO) following
TLR stimulation and cigarette smoke exposure, a condition which impairs glucocorticoid function. Methods
Human bronchial epithelial cells (BEAS2B) were preincubated with DEX (0.1nM-1µM) or RNO (0.1nM-1µM)
for 1h and then stimulated with the TLR2, 3, 4, 5 and 9 agonists peptidoglycan (10µg/ml), poly(I:C)
(10µg/ml), LPS (1µg/ml), flagellin (1µg/ml) and ODN-2395 (5µM), in the presence or absence of 1%
cigarette smoke extract (CSE) for 24h. IL-8 was measured by ELISA in cell supernatants. Results DEX
dose-dependently inhibited TLR2-, 3-, 4-, 5- and 9-induced IL-8 release in cells exposed to air (–logIC50 (M):
7.6, 8.0, 8.2, 8.7 and 7.7). CSE impaired the response of DEX following stimulation with TLR4, 5 and 9
agonists (–logIC50 (M): 6.4, 6.9 and 5.8) but not TLR2 and 3 agonists. In contrast, RNO dose-dependently
inhibited TLR2-, 3-, 4-, 5- and 9-induced IL-8 release in the absence (–logIC50 (M): 7.9, 8.0, 9.1, 8.1 and
7.4) and presence (–logIC50 (M): 7.8, 8.0, 9.0, 7.8 and 7.6) of CSE. Conclusions CSE exposure induced
glucocorticosteroid resistance in bronchial epithelial cells, as shown by the shift in the –logIC50 values of
DEX in inhibiting IL-8 release following TLR4, 5, and 9 stimulations compared to cells exposed to air. In
contrast, RNO inhibited IL-8 release induced by all TLR agonists tested in the presence or absence of CSE.