我国汉族Vogt—Koyanagi—Harada综合征患者人类白细胞抗原DQBI基因多态性研究  

Polymorphisms in HLA-DQB1 promoter and coding regions in Chinese Han patients with Vogt-Koyanagi-Harada syndrome

作　　者：闵寒毅[1] 刘英[2] 牛妮芳[2] 张美芬[1] 朱席琳[2] 赵家良[1] 

机构地区：[1]中国医学科学院北京协和医院眼科,100710 [2]中国协和医科大学冒家分子生物学重点实验室基础医学研究所

出　　处：《中华眼底病杂志》2010年第5期418-422,共5页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的 探讨人类白细胞抗原（HLA）DQB1启动子和编码子在Vogt-Koyanagi-Harada（VKH）综合征发病中的作用.方法 88例汉族VKH综合征患者和88名正常对照者纳入本研究.VKH综合征患者中,男性41例,女性47例,发病年龄15～67岁,平均年龄（36±11）岁.正常对照者中,男性42例,女性46例,年龄24～68岁,平均年龄（41±19）岁.采用聚合酶链式反应-序列特异性引物（PCR-SSP）方法进行HLA-DQB1等位基因分型,聚合酶链式反应-单链构象多态性（PCR-SSCP）-克隆-测序法检测HLA-DQB1基因启动子区（HLA-QBP）等位基因.应用x2检验或Fisher精确概率法进行统计分析.结果 VKH综合征患者中HLA-DQB1＊0401（0.318比0.045,x2=44.00,P=0.000,OR=9.8）和DQB1＊0303（0.068比0.006,x2=9.67,P=0.002,OR=12.81）频率明显高于正常对照者,差异有统计学意义.而HLA-DQB1＊0601（0.017比0.096,x2=10.39,P=0.001,OR=0.16）和HLA-DQB1＊0302（0.063比0.193,x2=13.48,P=0.000,OR=0.28）在VKH综合征患者中出现的频率显著低于正常对照者,差异有统计学意义.VKH综合征患者HLA-DQB1基因启动子区-189C/A的C等位基因频率显著高于正常对照者（0.324比0.074,x2=45.92,P=0.000）,而-227G/A等位基因频率低于正常对照者（0.011比0.108,x2=15.63,P=0.000）.VKH综合征患者中易感等位基因的组合（-189C和HLA-DQB1＊0401）频率明显高于正常对照者,正常对照者中抗性等位基因的组合（-227G和HLA-DQB1＊0601）频率显著高于VKH综合征患者.结论 VKH综合征发病可能与HLA-DQB1启动子和编码区相互作用有关.Objective To investigate the genetic interaction of HLA-DQB1 promoter and coding alleles in the pathogenesis of Vogt-Koyanagi-Harada syndrome （VKH）. Methods Eighty-eight Chinese Han patients with VKH and eighty-eight non-VKH normal controls were enrolled in this study. DNA was extracted from white blood cells of the subjects by phenol-chloroform method. Thirteen alleles were genotyped by polymerase chain reaction-sequence-specific primers （PCR-SSP）, polymerase chain reactionsingle strand conformation polymorphism （PCR-SSCP） and clone-sequencing was applied to determine the polymorphisms of the promoter and coding regions of HLA-DQB1 gene. Chromas and Bioedit software were used to analyze the sequences of the promoter of HLA-DQB1. Chi-square test and Fisherexact test were the statistical methods. Relationships among single nucleotide polymorphism （SNP） in the promoter and coding region were analyzed. Results Twelve of thirteen already known HLA-DQB1 alleles were genotyped by PCR-SSP in VKH patients. The most frequent allele in VKH patients was HLA-DQB1 ＊ 0401 （0.318,44.00, P=0.000, OR=9.8）. So was for HLA-DQB1＊0303 （0.068 vs. 0.006, x2=9.67, P=0.002,OR=12.81）. In contrast, the frequency of HLA-DQB1＊0601 （0.017 vs. 0.096, x2=10.39, P=0.001,OR=0.16） and HLA-DQB1 ＊ 0302 （0.062 vs. 0.193, x2=13.48, P=0.000, OR=0.28） in VKH patients were significantly lower than normal controls. Twelve SNP were found in all subjects. The frequency of C allele at position - 189C/A in VKH patients was significantly higher than that in controls （0.324 vs. 0.074, x2=45.92, P=0.000）. However, the frequency of G allele at position -227G/A in VKH patients was significantly lower than that in the normal controls （0.011 vs. 0.108, x2=15.63, P=0.000）. The frequency of combination of susceptible alleles in promoter and coding area （-189C and HLA-DQB1 ＊ 0401） in VKH patients was statistically higher than that in controls, the frequency of combination of resistant alleles in control （-227G and

关 键 词：葡萄膜脑膜脑炎综合征/病因学 HLA抗原/生理学 聚合酶链反应 

分 类 号：R773.9[医药卫生—眼科]