title: Simple diagnosis of STAT1 gain-of-function alleles in patients with chronic mucocutaneous candidiasis.
abstract: CMCD is a rare congenital disorder characterized by persistent or recurrent skin, nail, and mucosal membrane infections caused by Candida albicans. Heterozygous GOF STAT1 mutations have been shown to confer AD CMCD as a result of impaired dephosphorylation of STAT1. We aimed to identify and characterize STAT1 mutations in CMCD patients and to develop a simple diagnostic assay of CMCD. Genetic analysis of STAT1 was performed in patients and their relatives. The mutations identified were characterized by immunoblot and reporter assay using transient gene expression experiments. Patients' leukocytes are investigated by flow cytometry and immunoblot. Six GOF mutations were identified, three of which are reported for the first time, that affect the CCD and DBD of STAT1 in two sporadic and four multiplex cases in 10 CMCD patients from Japan. Two of the 10 patients presented with clinical symptoms atypical to CMCD, including other fungal and viral infections, and three patients developed bronchiectasis. Immunoblot analyses of patients' leukocytes showed abnormally high levels of pSTAT1 following IFN-γ stimulation. Based on this finding, we performed a flow cytometry-based functional analysis of STAT1 GOF alleles using IFN-γ stimulation and the tyrosine kinase inhibitor, staurosporine. The higher levels of pSTAT1 observed in primary CD14(+) cells from patients compared with control cells persisted and were amplified by the presence of staurosporine. We developed a flow cytometry-based STAT1 functional screening method that would greatly facilitate the diagnosis of CMCD patients with GOF STAT1 mutations.
authors.affiliation: 2.Hiroshima University Graduate School of Biomedical & Health Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan. masak@hiroshima-u.ac.jp.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Mizoguchi
authors.forename: Miyuki
authors.initials: M
authors.lastname: Tsumura
authors.forename: Satoshi
authors.initials: S
authors.lastname: Okada
authors.forename: Osamu
authors.initials: O
authors.lastname: Hirata
authors.forename: Shizuko
authors.initials: S
authors.lastname: Minegishi
authors.forename: Kohsuke
authors.initials: K
authors.lastname: Imai
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Hyakuna
authors.forename: Hideki
authors.initials: H
authors.lastname: Muramatsu
authors.forename: Seiji
authors.initials: S
authors.lastname: Kojima
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Ozaki
authors.forename: Takehide
authors.initials: T
authors.lastname: Imai
authors.forename: Sachiyo
authors.initials: S
authors.lastname: Takeda
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Okazaki
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Ito
authors.forename: Shin'ichiro
authors.initials: S
authors.lastname: Yasunaga
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Takihara
authors.forename: Vanessa L
authors.initials: VL
authors.lastname: Bryant
authors.forename: Xiao-Fei
authors.initials: XF
authors.lastname: Kong
authors.forename: Sophie
authors.initials: S
authors.lastname: Cypowyj
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Boisson-Dupuis
authors.forename: Anne
authors.initials: A
authors.lastname: Puel
authors.forename: Jean-Laurent
authors.initials: JL
authors.lastname: Casanova
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Morio
authors.forename: Masao
authors.initials: M
authors.lastname: Kobayashi
mesh_terms: D000483:Alleles; D002178:Candidiasis, Chronic Mucocutaneous / Q000235:genetics*; D004247:DNA / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D009154:Mutation; D010766:Phosphorylation; D050794:STAT1 Transcription Factor / Q000235:genetics* / Q000378:metabolism
chemical_list: D018950:Lipopolysaccharide Receptors; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D004247:DNA
keywords: CMCD; GOF; STAT1; pSTAT1