NADPH Oxidase Activity and Cytochromeb558Content of Human Epstein-Barr-Virus-Transformed B Lymphocytes Correlate with Expression of Genes Encoding Components of the Oxidase System

We investigated the NADPH oxidase activity, cytochromeb558content, and gene expression of gp91-phoxand p47-phoxin normal Epstein-Barr-virus (EBV)-transformed B lymphocytes, compared to EBV-transformed B lymphocytes from patients with X-linked chronic granulomatous disease (CGD), normal peripheral blood neutrophils or mononuclear cells, and the A301 or C8166 lymphoblastoid cell lines. CGD phenotypes included both “classic” disease with no detectable gp91-phoxprotein (termed X910) and “variant” phenotype with reduced but detectable gp91-phoxprotein (X91−). Normal EBV-transformed B lymphocytes show a dose-dependent PMA-induced superoxide release. Culturing these cells with IFN-γ (100 U/ml) and TNF-α (1000 U/ml), alone or in combination for 7 days, caused a modest increase in their NADPH oxidase activity (P> 0.05 in all situations). Normal EBV-transformed B lymphocytes have lower NADPH oxidase activity and cytochromeb558content than peripheral blood neutrophils or mononuclear cells (P< 0.05 in all situations). In contrast, they have higher NADPH oxidase activity and cytochromeb558content than X91−CGD EBV-transformed B lymphocytes (P< 0.05 in all situations). A301 or C8166 lymphoblastoid cell lines and X910CGD EBV-transformed B lymphocytes have barely detectable NADPH oxidase activity or cytochromeb558content (P< 0.05 in all situations). Gene expression studies also show a modest increase in expression and transcription rates of gp91-phoxand p47-phoxgenes in normal EBV-transformed B cells cultured with IFN-γ (100 U/ml) and TNF-α (1000 U/ml), alone or in combination for 7 days. We conclude that NADPH oxidase activity and cytochromeb558content correlate with gp91-phoxand p47-phoxgene expression in EBV-transformed B lymphocytes.