Essential role of nuclear factor-kappaB for NADPH oxidase activity in normal and anhidrotic ectodermal dysplasia leukocytes.
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.
Department of Pediatrics and Pharmacology, Center for Investigation in Pediatrics, State University of Campinas Medical School, Campinas, SP, Brazil.
Marcos
M
Luengo-Blanco
Carolina
C
Prando
Jacinta
J
Bustamante
Walmir Cutrim
WC
Aragão-Filho
Paulo Vitor Soeiro
PV
Pereira
Jussara
J
Rehder
Carolyn
C
Padden
Jean-Laurent
JL
Casanova
Peter E
PE
Newburger
Antonio
A
Condino-Neto
D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004476:Ectodermal Dysplasia / Q000276:immunology*; D015870:Gene Expression; D006105:Granulomatous Disease, Chronic / Q000473:pathology; D006801:Humans; D007962:Leukocytes / Q000378:metabolism* / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics*; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000502:physiology*; D010586:Phagocytes / Q000378:metabolism; D010587:Phagocytosis
D008562:Membrane Glycoproteins; D016328:NF-kappa B; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1