title: Rapid detection of intracellular p47phox and p67phox by flow cytometry; useful screening tests for chronic granulomatous disease.
abstract: Chronic granulomatous disease (CGD) is caused by defects of NADPH oxidase. The diagnosis of CGD can be made by analysis of NADPH oxidase activity, however, identification of the CGD subgroups is required before performing mutation analysis. The membrane-bound subunits, gp91phox and p22phox, can be quickly analyzed by flow cytometry, unlike the cytosolic components, p47phox and p67phox. We evaluated the feasibility of flow cytometric detection of p47phox and p67phox with specific monoclonal antibodies in two patients with p47phox deficiency and 7 patients with p67phox deficiency. Consistent with previous observations, p47phox and p67phox were expressed in phagocytes and B cells, but not in T or natural killer cells, from normal controls. In contrast, patients with p47phox and p67phox deficiency showed markedly reduced levels of p47phox and p67phox, respectively. These techniques will be useful to rapidly assess the expression of the cytosolic components, p47phox and p67phox, and represents important secondary screening tests for CGD.
authors.affiliation: Department of Pediatrics, School of Medicine, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, 920-8641, Japan. taizo@staff.kanazawa-u.ac.jp
authors.forename: Taizo
authors.initials: T
authors.lastname: Wada
authors.forename: Masahiro
authors.initials: M
authors.lastname: Muraoka
authors.forename: Tomoko
authors.initials: T
authors.lastname: Toma
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Imai
authors.forename: Tomonari
authors.initials: T
authors.lastname: Shigemura
authors.forename: Kazunaga
authors.initials: K
authors.lastname: Agematsu
authors.forename: Kohei
authors.initials: K
authors.lastname: Haraguchi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Moriuchi
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Oh-Ishi
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kitoh
authors.forename: Osamu
authors.initials: O
authors.lastname: Ohara
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Morio
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yachie
mesh_terms: D000293:Adolescent; D000328:Adult; D001402:B-Lymphocytes / Q000276:immunology*; D002469:Cell Separation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003600:Cytosol / Q000378:metabolism*; D004252:DNA Mutational Analysis; D005240:Feasibility Studies; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006105:Granulomatous Disease, Chronic / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008403:Mass Screening; D009154:Mutation / Q000235:genetics; D019255:NADPH Oxidases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D010586:Phagocytes / Q000276:immunology*; D010750:Phosphoproteins / Q000032:analysis*; D055815:Young Adult
chemical_list: D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1