Although Staphylococcus aureus is a major cause of pulmonary infection, the role played by this bacterium in the induction of inflammation of human airway epithelial cells (HAEC) is poorly understood. In this study, we investigated the inflammatory response of HAEC to S. aureus soluble virulence factors, and describe that the combination of the long-acting ₂-adrenergic receptor agonist (Salmeterol) with a glucocorticoid (Fluticasone Propionate) has an anti-inflammatory effect on HAEC. First, we demonstrate increased expression both at the mRNA and protein levels of the interleukin-8 (IL-8), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-) following incubation of HAEC in the presence of S. aureus soluble virulence factors, and the increase of i) the free nuclear factor-B (NF-B) and activator protein-1 (AP-1) activities, and ii) the phosphorylated (P-) extracellular signal-regulated kinases 1 and 2 (ERK1/ERK2), the P-c-Jun N-terminal kinase (JNK) and the P-isoform of the NF-B inhibitor (IB). Next, when HAEC were pre-incubated with the combination of Salmeterol and Fluticasone Propionate, the inflammatory response of HAEC was markedly attenuated in that levels of IL-8, IL-6, TNF-, NF-B, AP-1, P-ERK1/ERK2, P-JNK and P-IB decreased significantly. These data emphasize the deleterious effect of S. aureus soluble virulence factors and suggest that the combination of a ₂-adrenergic receptor agonist with a glucocorticoid may attenuate the associated airway epithelial inflammation.