中国汉族群体中异基因造血干细胞移植前人类白细胞抗原的高分辨分型

High-resolution typing of human leucocyte antigen before allogenic hematopoietic stem cell transplantation in Chinese Han people

目的:通过对异基因造血干细胞移植无关供受者对的人类白细胞抗原-A,B,DRB1基因的序列分析,探讨中国汉族群体中异基因造血干细胞移植前人类白细胞抗原高分辨分型的必要性。方法:实验于2000-08/2004-03在深圳市血液中心完成。供者和受者114对的血样分别来源中华骨髓库和各移植医院,供者和受者均签署知情同意书。采用聚合酶链式反应-测序分型技术对114对中国汉族无关供受者间的人类白细胞抗原-A,B,DRB1等位基因进行序列分析,观察中国汉族无关供受者对间等位基因水平的配合率、各等位基因家族的配对分布和错配分布规律。某些位点的模棱两可结果,则通过相应位点的高分辨聚合酶链式反应-序列特异性引物分型试剂进行分离并确认最后结果。结果:①高分辨与低分辨分型结果比较:114对无关供受者对中,110对人类白细胞抗原-A,B,DRB1的聚合酶链式反应-测序分型结果与低分辨(等位基因星号后两位数)DNA分型结果相吻合,4对聚合酶链式反应-测序分型结果与低分辨结果不相符。②供受者间人类白细胞抗原-A,B,DRB1高分辨配对分类结果:39%的中国汉族供受者对完全配合,但61%的供受者间存在至少一个等位基因的错配。③供受者间人类白细胞抗原-A,B,DRB1等位基因家族配对分布结果:3个座位的错配率分别为A(23%),B(10%),DRB1(17%);A和DRB1座位的错配比较集中,A座位仅见于A02,A11和A24家族,DRB1座位见于DRB104,DRB108,DRB112,DRB114和DRB115家族;而B座位的错配则比较分散,比较高的是B35,B48,B61和B62。④供受者间人类白细胞抗原-A,B,DRB1错配等位基因组合结果:3个座位的错配等位基因组合呈现较高的多样性,比较常见的几种组合为A0201/0207,A1101/1102,A0201/0206,A0203/0207,B4002/4006,DRB11201/1202和DRB11501/1502。结论:中国汉族异基因造血干细胞移植无关供受者对的人类白细胞抗原等位基因配合率低且其分布具有一定规律和特殊性,在移植前有必要对无关供受者对进行人类白细胞抗原高分辨分型。

AIM： To study the necessity of high-resolution typing of human leucocyte antigen before allogenic hematopoietic stem cell transplantation in Chinese Han people by sequencing human leucocyte antigen-A, B, DRB1 alleles of unrelated donor-recipient pairs of allogenic hematopoietic stem cell transplantation. METHODS： The experiment was conducted in Shenzhen Blood Center from August 2002 to March 2004. Blood samples of 114 donors and patients who had subscribed a consent form came from Chinese Bone Marrow Bank and associated hospitals, respectively. The human leucocyte antigen-A, B, and DRB1 alleles of 114 unrelated donor-recipient pairs in Chinese Han people were sequenced by polymerase chain reaction-sequence based typing technology so as to investigate the matched ratio of unrelated donorrecipient pairs in Chinese Han people at allele level, and the matched distribution and the mismatched regularity of allele families. Ambiguous resuits of some loci were isolated and confirmed by high-resolution polymerase chain reaction-sequence specific primer typing kits. RESULTS：①Comparison between results of high-resolution and low-resolution typing： Among the 114 unrelated donor-recipient pairs, results from 110 pairs of the human leucocyte antigen-A, B, DRB1 typed by polymerase chain reaction-sequence based on typing technology were found in accordance with low-resolution DNA typing （two numbers after the asterisk）, whereas 4 pairs out of accordance with low-resolution typing. ②Greup of human leucocyte antigen-A, B, DRB1 alleles between the donor-recipient pairs at high-resolution： Thirty-nine percent of donor-recipient pairs were fully matched and 61 percent mismatched at least one allele in Chinese Han people. ③Matched distribution of human leucocyte antigen-A, B, DRB1 allele families between the donor-recipient pairs： The mismatch ratio of A, B and DRB1 was 23%, 10% and 17%, respectively. Meanwhile, the mismatch at A and DRB1 loci were relatively centralized, only found at A＊02, A＊11 and A＊24 families of A locus, and at DRB1＊04, DRB1＊08, DRB1＊12, DRB1＊14 and DRB1＊15 of DRB1 locus; Whereas, the mismatch at B locus was relatively dispersive and the higher mismatches were B＊35, B＊48, B＊61 and B＊62. ④Mismatched allele combinations of human leucocyte antigen-A, B, DRB1 loci between the donor-recipient pairs： The mismatched allele combination of 3 loci showed higher diversity, and the most common mismatches were A＊0201/0207, A＊1101/1102, A＊0201/ 0206, A＊0203/0207, B＊4002/4006, DRB1＊1201/1202 and DRB1＊1501/ 1502. CONCLUSION： Human leucocyte antigen alleles between unrelated donor-recipient pairs of allogenic hematopoietic stem cell transplantation in Chinese Han people match lowly and distribute regularly and specially. It is concluded that it is necessary to perform high-resolution typing of human leucocyte antigen before transplantation.