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, Tzu Chi University, Hualien, Taiwan.
Tzu Chi Med J. 2016 Jul-Sep;28(3):99-102. doi: 10.1016/j.tcmj.2016.06.003. Epub 2016 Jul 29.
HLA-C04:82 is a low incidence HLA-C allele. The aim here is to report the ethnicity of C04:82 and its associated HLA haplotypes among Taiwanese individuals.
A sequence-based typing method was used to confirm this low incidence allele. Polymerase chain reaction was performed to amplify exons 2 and 3 of the HLA-A, HLA-B, and HLA-C loci and exon 2 of the HLA-DRB1 locus using group-specific primer sets. The amplicons were sequenced in both directions using Terminator Cycle Sequencing Ready Reaction kits and the manufacturer's protocols. The potential unrelated bone marrow stem cell donors in this study were randomized individuals with Taiwanese ethnicity who participated in the Tzu Chi Bone Marrow Donor Registry. The family members in the family part of the study were volunteer blood donors.
The DNA sequence of C04:82 was identical to C04:01:01:01 in exons 2, 3, and 4. It differed from C04:01:01:01 in exon 5 where a segment of nucleotides (CTAGCTGTC) was inserted between residues 969 and 970 of C04:01:01:01. The insertion of these nucleotides caused a 35 amino acid alteration to the protein sequence of C04:01:01:01. Three probable HLA haplotypes that were associated with C04:82 among Taiwanese individuals were deduced. Confirmation of the DNA and protein sequences of C*04:82 and its Taiwanese ethnicity were established in this study.
The ethnicity of the C04:82 allele and the deduced probable HLA haplotypes associated with the low-incidence C04:82 allele are of value for reference purposes for HLA testing laboratories. In addition, they can be used by search coordinators to aid the creation of a strategy for finding compatible stem cell donors for patients who carry this uncommon HLA allele.
HLA - C04:82是一种低发生率的HLA - C等位基因。本文旨在报告台湾人群中C04:82的种族分布及其相关的HLA单倍型。
采用基于序列的分型方法来确认这种低发生率的等位基因。使用组特异性引物对进行聚合酶链反应，以扩增HLA - A、HLA - B和HLA - C基因座的外显子2和3以及HLA - DRB1基因座的外显子2。使用终止循环测序预混反应试剂盒和制造商的方案对扩增产物进行双向测序。本研究中潜在的非亲属骨髓干细胞供体是随机选取的具有台湾种族的个体，他们参与了慈济骨髓捐赠登记处。研究中家庭部分的家庭成员是志愿献血者。
C04:82的DNA序列在外显子2、3和4上与C04:01:01:01相同。它在外显子5上与C04:01:01:01不同，在C04:01:01:01的第969和970位残基之间插入了一段核苷酸（CTAGCTGTC）。这些核苷酸的插入导致C04:01:01:01的蛋白质序列发生35个氨基酸的改变。推导出台湾人群中与C04:82相关的三种可能的HLA单倍型。本研究确定了C*04:82的DNA和蛋白质序列及其台湾种族特征。
C04:82等位基因的种族分布以及推导得出的与低发生率C04:82等位基因相关的可能HLA单倍型，对HLA检测实验室具有参考价值。此外，搜索协调员可利用它们来协助制定策略，为携带这种罕见HLA等位基因的患者寻找匹配的干细胞供体。
