NADPH oxidase activity and cytochrome b558 content of human Epstein-Barr-virus-transformed B lymphocytes correlate with expression of genes encoding components of the oxidase system.

Abstract

We investigated the NADPH oxidase activity, cytochrome b558 content, and gene expression of gp91-phox and p47-phox in 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-phox protein (termed X91(0)) and "variant" phenotype with reduced but detectable gp91-phox protein (X91(-)). Normal EBV-transformed B lymphocytes show a dose-dependent PMA-induced superoxide release. Culturing these cells with IFN-gamma (100 U/ml) and TNF-alpha (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 cytochrome b558 content than peripheral blood neutrophils or mononuclear cells (P < 0.05 in all situations). In contrast, they have higher NADPH oxidase activity and cytochrome b558 content than X91(-) CGD EBV-transformed B lymphocytes (P < 0.05 in all situations). A301 or C8166 lymphoblastoid cell lines and X91(0) CGD EBV-transformed B lymphocytes have barely detectable NADPH oxidase activity or cytochrome b558 content (P < 0.05 in all situations). Gene expression studies also show a modest increase in expression and transcription rates of gp91-phox and p47-phox genes in normal EBV-transformed B cells cultured with IFN-gamma (100 U/ml) and TNF-alpha (1000 U/ml), alone or in combination for 7 days. We conclude that NADPH oxidase activity and cytochrome b558 content correlate with gp91-phox and p47-phox gene expression in EBV-transformed B lymphocytes.

• Authors: A Condino-Neto, P E Newburger
• Journal: Archives of biochemistry and biophysics (Arch Biochem Biophys) Vol. 360 Issue 2 Pg. 158-64 (Dec 15 1998) ISSN: 0003-9861 [Print] United States
• PMID: 9851826 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
• Copyright: Copyright 1998 Academic Press.

Chemical References

• Cytochrome b Group
• Membrane Glycoproteins
• Phosphoproteins
• Tumor Necrosis Factor-alpha
• Superoxides
• Interferon-gamma
• cytochrome b558
• CYBB protein, human
• NADPH Oxidase 2
• NADPH Oxidases
• neutrophil cytosolic factor 1
• Tetradecanoylphorbol Acetate

Topics

• B-Lymphocytes (enzymology, metabolism)
• Blotting, Northern
• Cell Differentiation (drug effects)
• Cell Line, Transformed
• Cell Transformation, Viral
• Cytochrome b Group (metabolism)
• Gene Expression (drug effects)
• Herpesvirus 4, Human (physiology)
• Humans
• Interferon-gamma (pharmacology)
• Leukocytes, Mononuclear
• Membrane Glycoproteins (genetics)
• NADPH Oxidase 2
• NADPH Oxidases (genetics, metabolism)
• Neutrophils
• Phosphoproteins (genetics)
• Superoxides (metabolism)
• Tetradecanoylphorbol Acetate (pharmacology)
• Transcription, Genetic (drug effects)
• Tumor Necrosis Factor-alpha (pharmacology)