Clinical, Functional and Genetic Analysis of Twenty-Four Patients with Chronic Granulomatous Disease – Identification of Eight Novel Mutations in CYBB and NCF2 Genes
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Chronic granulomatous disease is an inherited disorder in which phagocytes lack a functional NADPH oxidase and cannot produce superoxide anions. The most common form is caused by mutations in CYBB encoding gp91phox. We investigated 24 CGD patients and their families. Twenty-one mutations in CYBB were classified as X910, X91+ or X91− variants according to cytochrome b 558 expression. Point mutations in encoding regions represented 50 % of the mutations found in CYBB, splice site mutations 27 %, deletions and insertions 23 %. Eight mutations in CYBB were novel leading to X910CGD cases. Two of these were point mutations: c493G>T and a double mutation c625C>G in exon 6 and c1510C>T in exon 12 leading to a premature stop codon at Gly165 in gp91phox and missense mutations His209Arg/Thr503Ile respectively. Two novel splice mutations in 5′intronic regions of introns 1 and 6 were found. A novel deletion/insertion c1024_1026delCTG/insT results in a frameshift introducing a stop codon at position 346 in gp91phox. The last novel mutation was the insertion of a T at c1373 leading to a frameshift and a premature stop codon at position 484 in gp91phox. For the first time the precise size of two large mutations in CYBB was determined by array-comparative genomic hybridization and carriers’ status were evaluated by multiplex ligation-dependent probe amplification assay. No clear correlation between clinical severity and CYBB mutations could be established. Of three mutations in CYBA, NCF1 and NCF2 leading to rare autosomal recessive CGD, one nonsense mutation c29G>A in exon 1 of NCF2 was new.
KeywordsChronic granulomatous disease NADPH oxidase Nox mutation
MJS is grateful for support from the University Joseph Fourier, Faculty of Medicine; the Ministry of Education and Research, MENRT; the Regional Clinical Research Department, DRCI, Grenoble University Hospital, the CGD Research Trust grant award reference J4G/09/09 and from the National Institutes of Health Grant N01–AI-30070 from the Immunodeficiency Network and the Primary Immunodeficiency Disease Consortium.
The authors are grateful to Dr D. Roos and Dr A. J. Verhoeven for the generous gift of mAb449 and mAb48 antibodies against the two subunits of the cytochrome b 558. We also thank Dr Alison Foote for editing the manuscript.
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