Occupational exposure by inhalation to vanadium-containing particles such as residual oil fly ash results in respiratory tract inflammation. This inflammation, characterized by abundant neutrophilia, appears to be initiated by alveolar macrophages (AMs) encountering particles and the subsequent release of proinflammatory cytokines. Intracellular signaling events in these cells in response to particles or their components are largely unknown. We investigated two immediate responses of AMs to vanadium exposure in vitro, the production of reactive oxygen intermediates (ROI) or respiratory burst (RB), and the tyrosine phosphorylation of cellular proteins. Macrophages exposed in vitro to 100 microM vanadyl chloride/1 microCi 48V incorporated 8.3% of the metal after 30 min. Exposure of AMs to increasing concentrations of sodium metavanadate resulted in a dose-dependent increase in production of ROI as measured by dichlorofluorescin oxidation. The lowest dose yielding a significant response was 50 microM, whereas 1000 microM increased RB activity by 173%. NADPH oxidase inhibitors deoxy-D-glucose (100 mM) and diphenylene iodonium (25 microM) reduced the metavanadate-induced RB by 62 and 71%, respectively, implicating NADPH oxidase as the primary cellular source of ROI. Enhanced cerium chloride oxidation in response to metavanadate localized to the plasma membrane consistent with increased NADPH oxidase activity. Pretreatment of AMs with the epidermal growth factor receptor inhibitor, tryphostin B50 (10 microM), reduced the metavanadate-induced RB, but did not influence overall tyrosine phosphorylation. Metavanadate and H2O2 exposure greatly increased overall tyrosine phosphorylation, yielding a similar but distinguishable pattern of phosphorylation in these cells. These observations demonstrate that in vitro metavanadate exposure regulates two distinct, yet related intracellular signaling pathways important in initiating inflammatory responses in these cells: (1) activation of the NADPH oxidase complex with subsequent increased ROI synthesis, and (2) enhanced tyrosine phosphorylation of cellular proteins.
Copyright 1999 Academic Press.