Yang Kuo-Liang
Laboratory of Immunogenetics, Tzu Chi Cord Blood Bank and Buddhist Tzu Chi Marrow Donor Registry, Buddhist Tzu Chi Stem Cells Centre, Hualien Tzu Chi Hospital, Hualien, Taiwan.
Department of Laboratory Medicine, Buddhist Tzu Chi University, Hualien, Taiwan.
Tzu Chi Med J. 2016 Jan-Mar;28(1):6-8. doi: 10.1016/j.tcmj.2016.02.001. Epub 2016 Mar 7.
HLA-DRB116:35 is a low incidence allele in the HLA-DRB1 locus. The objective of this study is to report the ethnicity of DRB116:35 and its deduced probable HLA associated haplotype in two Taiwanese unrelated bone marrow hematopoietic stem cell donors and to determine its variation from DRB116:02:01 and DRB116:01:01.
A sequence-based typing method was employed to confirm the low incidence allele DRB1*16:35. Polymerase chain reaction was performed to amplify exon 2 and exon 3 of the HLA-A and HLA-B loci and exon 2 of the HLA-DRB1 locus using group-specific primer sets. The amplicons were sequenced employing BigDye Terminator Cycle Sequencing Ready Reaction kits in both directions according to the manufacturer's protocols.
The DNA sequence of DRB116:35 is identical to DRB116:02:01 in exons 2, except for residue 364 where the C of DRB116:02:01 is replaced by the T of DRB116:35 (codon 93, CGG->TGG). The nucleotide exchange leads to an amino acid alteration to the protein sequence of DRB116:02:01 at residue 93 where the arginine (R) of DRB116:02:01 is changed to the tryptophan (W) of DRB116:35. We deduced the probable HLA haplotype in association with DRB116:35 in Taiwanese to be A11-B13- DRB1*16:35.
Information on the deduced probable HLA haplotype in association with the low incidence DRB1*16:35 allele that we report here is of value for HLA testing laboratories for reference purposes. In addition, it can be used by stem cell transplantation donor search coordinators to determine a strategy for finding compatible donors in unrelated bone marrow donor registries when a patient has this uncommon HLA allele.
HLA - DRB116:35是HLA - DRB1基因座中的一个低发生率等位基因。本研究的目的是报告两名台湾非亲属骨髓造血干细胞供体中DRB116:35的种族及其推导的可能的HLA相关单倍型,并确定其与DRB116:02:01和DRB116:01:01的差异。
采用基于序列的分型方法来确认低发生率等位基因DRB1*16:35。使用组特异性引物对进行聚合酶链反应,以扩增HLA - A和HLA - B基因座的外显子2和外显子3以及HLA - DRB1基因座的外显子2。根据制造商的方案,使用BigDye Terminator循环测序预混试剂盒对扩增子进行双向测序。
DRB116:35的DNA序列在第2外显子中与DRB116:02:01相同,除了第364位残基,DRB116:02:01的C被DRB116:35的T取代(密码子93,CGG->TGG)。这种核苷酸交换导致DRB116:02:01蛋白质序列在第93位残基处发生氨基酸改变,DRB116:02:01的精氨酸(R)变为DRB116:35的色氨酸(W)。我们推导台湾人群中与DRB116:35相关的可能的HLA单倍型为A11 - B13 - DRB1*16:35。
我们在此报告的与低发生率DRB1*16:35等位基因相关的推导的可能的HLA单倍型信息,对HLA检测实验室具有参考价值。此外,当患者具有这种罕见的HLA等位基因时,干细胞移植供体搜索协调员可利用该信息来确定在非亲属骨髓供体登记处寻找匹配供体的策略。