Plasmin regulates the activation of cell-associated latent TGF-beta(1) secreted by rat alveolar macrophages after in vivo bleomycin injury

Transforming growth factor beta s (TGF-beta s) are 25-kD multifunctional proteins that regulate inflammation and connective tissue synthesis. With rare exception TGF-beta(1) is secreted noncovalently bound to a latency-associated peptide (LAP) that renders the mature TGF-beta(1), biologically inactive. An important mechanism for the control of TGF-beta(1) action is the regulation of the post-translational processing that removes the LAP from the mature peptide and renders it biologically active. In a model of pulmonary inflammation and fibrosis induced by the antineoplastic antibiotic, bleomycin, we have demonstrated that explanted alveolar macrophages secrete progressively increasing quantities of a biologically active form of TGF-beta(1), the secretion of which was maximal 7 days after bleomycin administration. Thereafter, there was a rapid decline in the secretion of the active form of TGF-beta(1) whereas the latent form continued to be secreted in elevated quantities. Plasmin, a serine protease, was transiently generated by the same bleomycin-activated alveolar macrophages and paralleled the rise in active TGF-beta(1). When alpha(2)-antiplasmin, an inhibitor of plasmin, was added to cultures of alveolar macrophages, the post-translational activation of L-TGF-beta(1) was totally abrogated. When plasmin was added to alveolar macrophages in culture, there was complete activation of the L-TGF-beta(1) that had been secreted during the culture period. However, there was no effect of plasmin on the same alveolar macrophage-derived L-TGF-beta(1) in cell-free conditioned media. Our findings suggest that the secretion of an active form of TGF-beta(1) by alveolar macrophages is regulated by the generation of plasmin and requires that the alveolar macrophages be present. Because the diminution of active TGF-beta(1) coincides with the resolution of inflammation, this suggests that the availability of plasmin regulates the biologically active form of TGF-beta(1), and thus, the inflammation seen after bleomycin-induced lung injury.