广西造血干细胞骨髓库HLA-A、B、DRB1高分辨等位基因多态性研究

Polymorphism analysis of HLA-A,-B,-DRB1 alleles and haplotypes at high-resolution level in donors from marrow donor program registry of Guangxi

目的研究广西地区人群造血干细胞捐献志愿者的HLA-A、B、DRB1位点在高分辨水平上的等位基因多态性、基因频率及单体型分布特征。方法应用PCR-直接测序法(PCR-SBT)对中国造血干细胞捐献者资料库(CMDP)广西分库,3 431名无个体血缘关系健康供者的HLA-A、B、DRB1等位基因进行高分辨分型。结果共检出HLAA等位基因43个,B等位基因80个和DRB1等位基因42个,HLA-A-B-DRB1单体型检测到1507种。其中频率>0.05的常见等位基因,包括5种HLA-A∶A~*11∶01,A~*24∶02,A~*02∶03,A~*02∶07,A~*33∶03;6种HLA-B∶B~*46∶01,B~*40∶01,B~*15∶02,B~*58∶01,B~*13∶01,B~*38∶02;7种HLA-DRB1∶DRB1~*15∶01,DRB1~*16∶02,DRB1~*09∶01,DRB1~*03∶01,DRB1~*12∶02,DRB1~*14∶54,DRB1~*15∶02。487条HLA-A-B单体型中,79条单体型呈现显著的正连锁不平衡(ALD>0,HF≥1.09×10-4,χ2>3.84),5条表现为强连锁不平衡(RLD>0.40),18条单体型的频率>0.01;597条HLA-B-DRB1单体型中,68条单体型呈现显著的正连锁不平衡(ALD>0,HF≥1.09×10-4,χ2>3.84),8条表现为强连锁不平衡(RLD>0.40),16条单体型的频率高于0.01;1 507条HLA-A-B-DRB1单体型中,统计分析三座位单体型最常见的是A~*33∶03-B~*58∶01-DRB1~*03∶01(0.058 8)。结论广西地区人群HLA-A、B、DRB1位点在高分辨水平上获得等位基因频率、单体型分布数据及相关遗传参数,为今后开展临床造血干细胞移植、中国人群CWD表的制订和骨髓库最适库容的评估等研究及相关疾病研究提供本地区信息。

Objective To analyze the genetic polymorphism,allele frequencies and the distribution of haplotypes of HLA-A,B and DRB1 alleles by analyzing the hematopoietic stem cell donors（ HSC） in native population of Guangxi. Methods Polymerase chain reaction-sequence-based typing method（ PCR-SBT） was conducted for HLA-A,B and DRB1 highresolution genotyping on 3 431 unrelated healthy hematopoietic stem cell donors from Guangxi sub-registry of Chinese National Marrow Donor Program（ CMDP）. Results A total of 43 HLA-A alleles,80 B alleles,and 42 DRB1 alleles were detected,and 1 507 A-B-DRB1 haplotypes were statistically inferred. Of these,high frequency alleles found at a frequency of ≥5%were 5 HLA-A（ A^＊11∶ 01,A^＊24∶ 02,A^＊02∶ 03,A^＊02∶ 07,A^＊33∶ 03）,6 HLA-B（ B^＊46∶ 01,B^＊40∶ 01,B^＊15∶ 02,B^＊58∶ 01,B^＊13∶ 01,B^＊38 ∶ 02）,7 HLA-DRB1（ DRB1^＊15 ∶ 01,DRB1^＊16 ∶ 02,DRB1^＊09 ∶ 01,DRB1^＊03 ∶ 01,DRB1^＊12 ∶02,DRB1^＊14∶ 54,DRB1^＊15∶ 02）. Among 487 kinds of HLA-A-B haplotypes,there were 18 kinds of frequencies that were higher than 0. 01,and 79 kinds with statistically significant and positive linkage disequilibrium（ ALD〉 0,HF acuity 1. 09× 10^4,χ^2〉 3. 84）. Moreover,among 579 kinds of the HLA-B-DR haplotypes,there were 16 kinds of frequencies higher than 0. 01,and 68 kinds with statistically significant and positive linkage disequilibrium. Statistical analysis of the three-locus haplotypes showed that A^＊33∶ 03-B^＊58∶ 01-DRB1^＊03∶ 01（0. 058 8） was the most predominant. Conclusion Gene frequencies and haplotypic associations within HLA-A,-B,and-DRB1 loci were determined at a high-resolution allelic level in native population of Guangxi and should provide useful native information on the correlation of HLA with bone marrow donor registry,composition of the CWD alleles in the Chinese Han population,and estimation of optimal bone marrow registry size and disease association studies.