Modulation of macrophage activation by ammonium metavanadate

The molecular basis of the immunotoxic effect of ammonium metavanadate on signal transduction involved in macrophage activation was studied in resident peritoneal macrophages (PEM) and a murine macrophage-like cell line, J774. A fourfold elevation in cytosolic free calcium levels was observed within 10 s following lipopolysaccharide (LPS) stimulation of the non-vanadate-exposed controls both in vitro and in vivo; the levels returned to prestimulation values within 70 s. Exposure to phorbol ester (PMA) did not result in any appreciable change in cytosolic free calcium levels. Compared to untreated controls, treatment with vanadate caused a significant elevation in basal cytosolic calcium levels. Such elevation was not enhanced further by LPS. LPS stimulation of macrophages also resulted in a significant elevation of membrane-associated protein kinase C (PKC) activity, which was, however, inhibited in a dose-dependent manner by vanadate in both in vitro and in vivo studies. Exposure to PMA also resulted in a significant elevation of membrane-associated PKC activity; vanadate treatment at lower levels did not cause downregulation, indicating that vanadate at these levels interfered with the receptor-mediated events rather than the enzyme directly. Vanadate at higher exposure levels inhibited the activity even in PMA-stimulated macrophages. No significant difference occurred in cytosolic PKC activities in control macrophages; vanadate treatment at lower levels resulted in a significant elevation of cytosolic PKC activities following stimulation with LPS or PMA, indicating that vanadate might be interfering with the translocation process.