J Korean Med Sci.  2009 Dec;24(6):1045-1050. 10.3346/jkms.2009.24.6.1045.

Genetic Analysis of 10 Unrelated Korean Families with p22-phox-deficient Chronic Granulomatous Disease: An Unusually Identical Mutation of the CYBA Gene on Jeju Island, Korea

• Kim YM
• Park JE
• Kim JY
• Lim HK
• Nam JK
• Cho M
• Shin KS

• KMID: 1783131
• DOI: http://doi.org/10.3346/jkms.2009.24.6.1045

Abstract

Chronic granulomatous disease (CGD) is a rare hereditary disorder characterized by recurrent life-threatening bacterial and fungal infections. The underlying defect in CGD is an inability of phagocytes to produce reactive oxygen species as a result of defects in NADPH oxidase. Considering that CGD generally affects about 3-4 in 1,000,000 individuals, it is surprising that the prevalence of CGD on Jeju Island is 20.7 in 1,000,000 individuals. We performed genetic analysis on 12 patients from 10 unrelated families and found that all patients had an identical homozygous single-base substitution of C to T in exon 1 (c.7C>T) of the CYBA gene, which was expected to result in a nonsense mutation (p.Q3X). Because Jeju Island has long been a geologically isolated region, the high prevalence of CGD on Jeju Island is presumably associated with an identical mutation inherited from a common ancestor or proband.

Keyword

Granulomatous Disease, Chronic; CYBA; NADPH Oxidase; Molecular Diagnosis; Codon, Nonsense; Korea

MeSH Terms

Adolescent
Animals
Asian Continental Ancestry Group/*genetics
Base Sequence
Child
Child, Preschool
DNA Mutational Analysis
Female
Geography
Granulomatous Disease, Chronic/*genetics
Humans
Infant
Korea
Male
Molecular Sequence Data
*Mutation
NADPH Oxidase/*genetics
Pedigree
Young Adult

Figure

• 

  Fig. 1 Dihydrorhodamine-1,2,3 (DHR) flow cytometric analysis of granulocyte oxidative activity in family III. There was no difference in phagocyte oxidative activity between the parents of patient 4 and healthy controls after no stimulation (black) or stimulation with PMA (red). A lack of phagocyte oxidative activity after stimulation with PMA was observed in patient 4.

• 

  Fig. 2 Western blot analysis of NADPH oxidase components in family III. Western blot analysis demonstrated the absence of p22-phox expression in patients 4 and 5, but the expression of gp91phox, p47-phox, p67phox, and p40-phox were normal. There was no abnormal expression of any NADPH oxidase components in other family members. F, Father of patients 4 and 5; M, Mother of patients 4 and 5; P4, Patient 4; S, Elder sister of patient 5; P5, Patient 5; B, Younger brother of patient 5.

• 

  Fig. 3 RT-PCR of CYBA cDNA from family III. The mRNA of p22-phox was reverse transcribed as described above. There was no difference in size between the p22-phox cDNAs in family III and those in healthy controls. F, Father of patients 4 and 5; M, Mother of patients 4 and 5; P4, Patient 4; S, Elder sister of patient 5; P5, Patient 5; B, Younger brother of patient 5.

• 

  Fig. 4 Sequence analysis of the CYBA gene from family III. (A) Pedigree of family III. (B) Sequence analysis of the CYBA gene from family III. Patients 4 and 5 had an identical, homozygous, single-base substitution of C- to T in exon 1 (c.7C>T), which was expected to result in a nonsense mutation (p.Q3X). The sequencing results for other family members showed a double signal at the same position, indicating that they were heterozygous for the mutation. *site of mutation. F, Father of patients 4 and 5; M, Mother of patients 4 and 5; P4, Patient 4; S, Elder sister of patient 5; P5, Patient 5; B, Younger brother of patient 5.

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