TY - JOUR
T1 - Simple diagnosis of STAT1 gain-of-function alleles in patients with chronic mucocutaneous candidiasis
AU - Mizoguchi, Yoko
AU - Tsumura, Miyuki
AU - Okada, Satoshi
AU - Hirata, Osamu
AU - Minegishi, Shizuko
AU - Imai, Kohsuke
AU - Hyakuna, Nobuyuki
AU - Muramatsu, Hideki
AU - Kojima, Seiji
AU - Ozaki, Yusuke
AU - Imai, Takehide
AU - Takeda, Sachiyo
AU - Okazaki, Tetsuya
AU - Ito, Tsuyoshi
AU - Yasunaga, Shinich'iro
AU - Takihara, Yoshihiro
AU - Bryant, Vanessa L.
AU - Kong, Xiao Fei
AU - Cypowyj, Sophie
AU - Boisson-Dupuis, Stéphanie
AU - Puel, Anne
AU - Casanova, Jean Laurent
AU - Morio, Tomohiro
AU - Kobayashi, Masao
PY - 2014/4
Y1 - 2014/4
N2 - 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.
AB - 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.
KW - CMCD
KW - GOF
KW - STAT1
KW - pSTAT1
UR - http://www.scopus.com/inward/record.url?scp=84899791876&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84899791876&partnerID=8YFLogxK
U2 - 10.1189/jlb.0513250
DO - 10.1189/jlb.0513250
M3 - Article
C2 - 24343863
AN - SCOPUS:84899791876
SN - 0741-5400
VL - 95
SP - 667
EP - 676
JO - Journal of Leukocyte Biology
JF - Journal of Leukocyte Biology
IS - 4
ER -