TY - JOUR T1 - Anti-inflammatory effects of roflumilast N-oxide and dexamethasone in human bronchial epithelial cells stimulated with toll-like receptor agonists JF - European Respiratory Journal JO - Eur Respir J VL - 42 IS - Suppl 57 SP - P3128 AU - Javier Milara AU - Javier Lluch AU - Patricia Almudever AU - Teresa Peiró AU - Adela Serrano AU - Magdalena Alonso-Galicia AU - Jose Freire AU - Xiaozhong Qian AU - Julio Cortijo Y1 - 2013/09/01 UR - http://erj.ersjournals.com/content/42/Suppl_57/P3128.abstract N2 - ObjectiveBacterial 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.MethodsHuman 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.ResultsDEX 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.ConclusionsCSE 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. ER -