The glucocorticoid receptor (GR) is a ubiquitously expressed transcription factor present in most cell types. Upon ligand binding, the GR is activated and translocates into the nucleus where it transmits the anti-inflammatory actions of glucocorticoids. Here, we describe the ligand-independent activation of GR by the ␤ 2 -adrenergic receptor (␤ 2 -AR) agonists, salbutamol and salmeterol, in primary human lung fibroblasts and vascular smooth muscle cells. Immunohistochemistry demonstrated expression of GR and the ␤ 2 -AR by fibroblasts and vascular smooth muscle cells. Treatment of the cells with the ␤ 2 -AR agonists, salbutamol or salmeterol, resulted in translocation of GR into the nucleus beginning at 30 min, as shown by immunohistochemistry and Western blotting of cytosolic and nuclear cell extracts. In comparison, activation of GR induced by the corticosteroids dexamethasone and fluticasone occurred at the same time after treatment (30 min) but resulted in a more complete depletion of GR from the cytosolic compartment. Electrophoretic mobility shift assays confirmed that nuclear GR, activated by both ␤ 2 -AR agonists and glucocorticoids, actively bound to the GR consensus sequence (GR element). Functional activation of the GR was confirmed by a Luciferase reporter gene assay, using a GR driven promoter fragment from the p21 (WAF1/CIP1) gene. The effects of the ␤ 2 -AR agonists, salbutamol and salmeterol, were dependent upon binding to the ␤ 2 -AR, because blocking of ␤ 2 -AR with propranolol abrogated GR activation. GR activation appeared to involve cAMP. In summary, these data show that ␤ 2 -AR agonists are potent activators of GR. Ligandindependent activation of GR by ␤ 2 -AR agonists may substantially mediate the anti-inflammatory actions of these drugs observed in vitro and in vivo.