摘要目的 探讨中国南方汉族造血干细胞捐献者人类白细胞抗原(HLA)-A、-B、-C、-DRB1及-DQB1位点高分辨等位基因的多态性分布.方法 选择2016年1月至2017年12月于中国造血干细胞捐献者资料库(CMDP)深圳分库登记的4 262例中国南方汉族造血干细胞捐献者为研究对象.采用PCR-序列特异性寡核苷酸探针(SSO)和二代测序(NGS)技术对本研究4 262例造血干细胞捐献者的HLA-A、-B、-C、-DRB1及-DQB1位点进行高分辨等位基因分型检测.采用直接计数法分别计算上述5个HLA位点的常见、确认及罕见等位基因频率.采用x2检验判断研究对象的HLA-A、-B、-C、-DRB1及-DQB1各位点等位基因的分布是否符合Hardy-Weinberg平衡定律.结果 ①本研究4 262例中国南方汉族造血干细胞捐献者的HLA-A、-B、-C、-DRB1及-DQB1位点等位基因观察值与期望值分别比较,差异均无统计学意义(x2 =295.068、572.482、355.205、362.584、312.150,P＞0.05).本研究4 262例造血干细胞捐献者的HLA-A、-B、-C、-DRB1及-DQB1位点基因型分布均符合Hardy-Weinberg平衡定律.②本研究4 262例中国南方汉族造血干细胞捐献者中,检出的HLA-A、-B、-C、-DRB1及-DQB1位点的等位基因分别为49、97、37、48和22种,其中等位基因频率＞0.05的常见等位基因包括:HLA-A* 11∶01、24∶02、02∶07、02∶01、33∶03和02∶03,累计等位基因频率为0.767 1;HLA-B* 40∶01、46∶01、58∶01、13∶01和51∶01,累计等位基因频率为0.471 6;HLA-C* 01∶02、07∶02、03∶04、08∶01、03∶02、06∶02和03∶03,累计等位基因频率为0.782 5;HLA-DRB1* 09∶01、15∶01、12∶02、07∶01、08∶03、03∶01、11∶01和04∶05,累计等位基因频率为0.656 4;以及HLA-DQB1* 03∶01、03∶03、06∶01、05∶02、03∶02、02∶01、05∶03、06∶02和02∶02,累计等位基因频率为0.877 6.③本研究4 262例中国南方汉族造血干细胞捐献者中,HLA-A、-B、-C、-DRB1及-DQB1位点的确认及罕见等位基因分别检出22、42、14、11和7种,其中HLA-A* 29∶03,HLA-B* 18∶04、15∶296和54∶03,HLA-C* 12∶12、HLA-DRB1* 04∶101,以及HLA-DQB1*02∶10为罕见等位基因,目前尚未被收录于CMDP的《常见及确认等位基因表(CWD)》2.3版本.结论 本研究获得较为完整的中国南方汉族造血干细胞捐献者的HLA-A、-B、-C、-DRB1及-DQB1位点的高分辨等位基因多态性分布数据,记录并统计HLA确认及罕见等位基因.本研究结果有助于CMDP中HLA基因人群分布和多态性数据的积累,并且为完善CMDP的CWD提供参考依据.

abstractsObjective To explore the human leukocyte antigen (HLA)-A,-B,-C,-DRB1 and-DQB1 high-resolution alleles polymorphism in hematopoietic stem cells donors of Chinese Han population from Southern China.Methods A total of 4 262 cases hematopoietic stem cell donors of chinese Han population in Shenzhen China Marrow Donor Program (CMDP) from January 2016 to December 2017 were selected as subjects.HLA-A,-B,-C,-DRB1 and-DQB1 loci for high resolution typing of these 4 262 cases hematopoietic stem cell donors were detected by PCR-sequence-specific oligonucleotide probes (SSO) and next-generation sequencing (NGS) analysis.The frequencies of common,well-documented and rare alleles were studied by direct counting method.The x2 test was used to determine whether the distribution of HLA-A,-B,-C,-DRB1 and-DQB1 alleles of these 4 262 cases hematopoietic stem cell donors accorded with the Hardy-Weinberg equilibrium.Results ① Compared observed values of HLA-A,-B,-C,-DRB1 and-DQB1 alleles in these 4 262 cases hematopoietic stem cell donors of Chinese Han population from Southern China with the expected values,there were not statistically significance (x2 =295.068,572.482,355.205,362.584,312.150;P＞0.05),which indicated that the distribution of HLA-A,-B,-C,-DRB1 and-DQB1 alleles of all hematopoietic stem cell donors in this study conformed with Hardy-Weinberg equilibrium.② The alleles of HLA-A,-B,-C,-DRB1 and-DQB1 loci in 4 262 cases hematopoietic stem cells donors of Chinese Han population from Southern China were 49,97,37,48 and 22,respectively.Among them,the common alleles with frequencies higher than 0.05 included HLA-A * 11 ∶ 01,24∶ 02,02∶07,02∶01,33∶03 and 02∶03;HLA-B*40∶01,46∶01,58∶01,13∶01 and 51∶01;HLA-C*01∶02,07∶02,03∶04,08∶01,03∶02,06∶02 and 03∶03;HLA-DRB1*09∶01,15∶01,12∶02,07∶01,08∶03,03∶01,11∶01 and 04∶05;and HLA-DQB1*03∶01,03∶03,06∶01,05∶02,03∶02,02∶01,05∶03,06∶02 and 02∶02.For each of the locus,the cumulative allele frequencies of common alleles were 0.767 1,0.471 6,0.782 5,0.656 4 and 0.877 6,respectively.③ The well-documented and rare alleles of HLA-A,-B,-C,-DRB1 and-DQB1 loci were detected as 22,42,14,11 and 7.Among them,the rare alleles of HLA-A* 29∶03;HLA-B* 18∶04,15∶296 and 54∶03;HLA-C * 12∶12;HLA-DRB1 * 04∶101 and HLA-DQB1*02∶10 were not included in the Common Alleles and Well-Documented Alleles (CWD) v2.3 of CMDP.Conclusions Relatively complete high-resolution allelic distribution data of HLA-A,-B,-C-DRB1 and-DQB1 loci in hematopoietic stem cells donors of Chinese Han population from Southern China were obtained in this study.And HLA well-documented and rare alleles were recorded.The results of this study contribute to the accumulation on population distribution and polymorphism data of HLA gene in hematopoietic stem cell donors,and provide reference for improving CWD of CMDP.