Molecular basis of interferon-gamma and lipopolysaccharide enhancement of phagocyte respiratory burst capability. Studies on the gene expression of several NADPH oxidase components.

ABSTRACT

In this study, we analyzed the expression of genes encoding for components of the phagocyte superoxide anion-generating system in human phagocytes treated with interferon-gamma (IFN-gamma) or lipopolysaccharide (LPS). Human neutrophils express high levels of the 47-kDa cytosolic factor (p47-phox), which are down-regulated after treatment with IFN-gamma, but not with LPS. On the contrary, the steady-state levels of the heavy chain subunit of cytochrome b558 (gp91-phox) were increased by IFN-gamma and LPS in human monocyte-derived macrophages and neutrophils in a time- and dose-dependent fashion, whereas cytochrome b558 light chain subunit (p22-phox) mRNA was not influenced by either agent. Studies on post-transcriptional regulation at the level of mRNA stability indicate that, in neutrophils, IFN-gamma has no influence on gp91-phox and p47-phox mRNA half-lives. The content of the two cytochrome b558 subunits was quantified by enzyme-linked immunosorbent assay, which revealed that, in neutrophils, gp91-phox levels doubled after 4 h of treatment with IFN-gamma or LPS. Monocyte/macrophage maturation was associated with a gradual decrease in gp91-phox mRNA and protein levels, which were both restored by treatment with IFN-gamma for 24-48 h. These results suggest that induction of the gp91-phox gene and protein product by IFN-gamma or LPS is an important requirement in the mechanism of the enhancement of neutrophil and macrophage oxidative metabolism.