Essential role of nuclear factor-kappaB for NADPH oxidase activity in normal and anhidrotic ectodermal dysplasia leukocytes.

Abstract

This work investigated the functional role of nuclear factor-kappaB (NF-kappaB) in respiratory burst activity and in expression of the human phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase genes CYBB, CYBA, NCF1, and NCF2. U937 cells with a stably transfected repressor of NF-kappaB (IkappaBalpha-S32A/S36A) demonstrated significantly lower superoxide release and lower CYBB and NCF1 gene expression compared with control U937 cells. We further tested Epstein-Barr virus (EBV)-transformed B cells from patients with anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID), an inherited disorder of NF-kappaB function. Superoxide release and CYBB gene expression by EDA-ID cells were significantly decreased compared with healthy cells and similar to cells from patients with X-linked chronic granulomatous disease (X91(0) CGD). NCF1 gene expression in EDA-ID S32I cells was decreased compared with healthy control cells and similar to that in autosomal recessive (A47(0)) CGD cells. Gel shift assays demonstrated loss of recombinant human p50 binding to a NF-kappaB site 5' to the CYBB gene in U937 cells treated with NF-kappaB inhibitors, repressor-transfected U937 cells, and EDA-ID patients' cells. Zymosan phagocytosis was not affected by transfection of U937 cells with the NF-kappaB repressor. These studies show that NF-kappaB is necessary for CYBB and NCF1 gene expression and activation of the phagocyte NADPH oxidase in this model system.

• Authors: Marcos Luengo-Blanco, Carolina Prando, Jacinta Bustamante, Walmir Cutrim Aragão-Filho, Paulo Vitor Soeiro Pereira, Jussara Rehder, Carolyn Padden, Jean-Laurent Casanova, Peter E Newburger, Antonio Condino-Neto
• Journal: Blood (Blood) Vol. 112 Issue 4 Pg. 1453-60 (Aug 15 2008) ISSN: 1528-0020 [Electronic] United States
• PMID: 18523147 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Membrane Glycoproteins
• NF-kappa B
• CYBB protein, human
• NADPH Oxidase 2
• NADPH Oxidases
• neutrophil cytosolic factor 1

Topics

• Cell Line, Transformed
• Cells, Cultured
• Ectodermal Dysplasia (immunology)
• Gene Expression
• Granulomatous Disease, Chronic (pathology)
• Humans
• Leukocytes (metabolism, pathology)
• Membrane Glycoproteins (genetics)
• NADPH Oxidase 2
• NADPH Oxidases (genetics, metabolism)
• NF-kappa B (physiology)
• Phagocytes (metabolism)
• Phagocytosis