Plasminogen-stimulated inflammatory cytokine production by airway smooth muscle cells is regulated by annexin A2

Plasminogen has a role in airway inflammation. Airway smooth muscle (ASM) cells cleave plasminogen into plasmin, a protease with proinflammatory activity. In this study, the effect of plasminogen on cytokine production by human ASM cells was investigated in vitro. Levels of IL-6 and IL-8 in the medium of ASM cells were increased by incubation with plasminogen (5?50 ?g/ml) for 24 hours (P <0.05; n = 6?9), corresponding to changes in the levels of cytokine mRNA at 4 hours. The effects of plasminogen were attenuated by a2-antiplasmin (1 ?g/ml), a plasmin inhibitor (P <0.05; n = 6?12). Exogenous plasmin (5?15 mU/ml) also stimulated cytokine production (P <0.05; n = 6?8) in a manner sensitive to serine-protease inhibition by aprotinin (10 KIU/ml). Plasminogen-stimulated cytokine production was increased in cells pretreated with basic fibroblast growth factor (300 pM) in a manner associated with increases in urokinase plasminogen activator expression and plasmin formation. The knockdown of annexin A2, a component of the putative plasminogen receptor comprised of annexin A2 and S100A10, attenuated plasminogen conversion into plasmin and plasmin-stimulated cytokine production by ASM cells. Moreover, a role for annexin A2 in airway inflammation was demonstrated in annexin A2-/- mice in which antigen-induced increases in inflammatory cell number and IL-6 levels in the bronchoalveolar lavage fluid were reduced (P <0.01; n = 10?14). In conclusion, plasminogen stimulates ASM cytokine production in a manner regulated by annexin A2. Our study shows for the first time that targeting annexin A2?mediated signaling may provide a novel therapeutic approach to the treatment of airway inflammation in diseases such as chronic asthma.