人类白细胞抗原A和DRB1基因与1型糖尿病的相关性研究

目的研究江苏地区人类白细胞抗原（HL4）-A和月出-DRBl基因频率及单倍型与1型糖尿病的相关性。方法于2008至2009年在江苏省江阴市和南京市选择自发酮症急性起病、依赖胰岛素治疗的1型糖尿病门诊或住院患者51例（1型糖尿病组），其中男27例，女24例，平均发病年龄（14±9）岁。另以2002至2006年登记加入中华骨髓库江苏分库的汉族无关骨髓捐献志愿者HLA分型资料20248份为对照。从受试者外周血中提取细胞基因组DNA，利用聚合酶链反应-寡核苷酸探针序列特异性引物技术进行HLA—A和月似-DRBl基因分型，应用Arlequin软件分析计算单倍型频率及连锁不平衡情况，通过卡方检验等比较组间基因频率、单倍型频率及疾病相关性分析。结果HLA-A点频率最高的为A＊02和A＊24，HLA-DRBl位点频率最高的为DRBl＊09。与对照组相比，1型糖尿病组HLA—A＊3[6．86％（7／102）比2．43％（984／40496），X^2=8．40，P〈0．01，比值比（OR）=3．00]、A＊24[27．45％（28／102）比16．79％（6799／40496），X^2=8．27，P〈0．01，OR=1．87]和DRBl＊09[32．35％（33／102）比16．15％（6540／40496），X^2=19．69，P〈0．01，OR=2．47]、DRBl＊03[15．69％（16／102）比4．76％（1927／40496），X^2=26．66，P〈0．01，OR=3．69]频率显著升高。A＊01-DRBl＊03在连锁不平衡分析中为1型糖尿病组所特有。结论检测出位点A＊03、A＊24及DR03、DR09对1型糖尿病的易患性，发现3个有易患作用的A—DR单倍型，观察到与1型糖尿病关联的祖先单倍型A01DR03。

Relationship between human leukocyte antigen A and DRB1 gene and type 1 diabetes mellitus

Objective To investigate the relationship between type l diabetes mellitus and human leukocyte antigen （ HLA ） -el and-DRBl gene frequencies and haplotypes. Methods Fifty-one patients （ male 27, female 24, average age of onset （ 14±9 ） y） with spontaneously acute ketotic onset and insulin-dependent type 1 diabetes mellitus （ T1DM ） were enrolled during 2008 and 2009 from Jiangyin and Nanjing city in Jiangsu Province. HLA typing data with the similar genetic background of 20 248 unrelated donors in Jiangsu Branch of Chinese Marrow Donor Progranl were set as controls. DNA was extracted from the peripheral blood, and HLA-A or DRB1 gene was genotyped with polymerase chain reaction-sequence specific oligonueleotide primers, and the haplotype frequencies, linkage disequilibrium and cmxelation with type 1 diabetes were also analyzed by Arlequin software and Chi-square test in the two groups. Results The frequencies of A ＊ 02 and A ＊ 24 were the highest in HLA-A locus, and DRBI ＊ 09 was the highest in DRB1 locus. Compared with the controlled group, the frequencies of HLA-A ＊ 03, HLA-A ＊ 24, DRBI ＊ 09 and DRB1 ＊ 03 were significantly increased in T1DM patients（6. 86% vs 2.43% ,27.45% vs 16. 79% ,32.35% vs 16. 15% , 15.69% vs 4. 76% , respectively; X^2 = 8.40,8.27,19.69,26. 66; odds ratio = 3.00, 1.87, 2.47,3.69;all P 〈 0.01 ） . A ＊ 01-DRB1 ＊ 03 haplotype was specific with T1DM group in linkage disequilibrium analysis. Conclusion We find the herd susceptibility of loci A ＊ 03, A ＊ 24 and DR03, DR09 in T1DM subjects, and three A-DR susceptibility haplotypes. The relationship between ancestor haplotype of A01DR03 and T1DM is confirmed.