Blood Center of Zhejiang Province, Institute of Transfusion Medicine, Hangzhou, China.
Key Laboratory of Blood Safety Research of Zhejiang Province, Hangzhou, China.
Front Immunol. 2021 Nov 12;12:755661. doi: 10.3389/fimmu.2021.755661. eCollection 2021.
RhD is the most important and complex blood group system because of its highly polymorphic and immunogenic nature. RhD variants can induce immune response by allogeneic transfusion, organ transplantation, and fetal immunity. The transfusion strategies are different for RhD variants formed by various alleles. Therefore, extensive investigation of the molecular mechanism underlying RhD variants is critical for preventing immune-related blood transfusion reactions and fetal immunity.
RhD variants were collected from donors and patients in Zhejiang Province, China. The phenotypes were classified using the serologic method. The full coding regions of gene were analyzed using the PCR-SBT method. The multiplex ligation-dependent probe amplification (MLPA) assay was used to analyze the genotype and gene copy number. SWISS-MODLE and PyMOL software were used to analyze 3D structures of RhD caused by the variant alleles. The effect of non-synonymous substitutions was predicted using Polymorphism Phenotyping algorithm (PolyPhen-2), Sorting Intolerant From Tolerant (SIFT), and Protein Variation Effect Analyzer (PROVEAN) software.
In the collected RhD variants, 28 distinct variant alleles were identified, including three novel variant alleles. RH-MLPA assay is advantageous for determining the copy number of gene. 3D homology modeling predicted that protein conformation was disrupted and may explain RhD epitope differential expression. A total of 14 non-synonymous mutations were determined to be detrimental to the protein structure.
We revealed the diversity of alleles present in eastern Chinese RhD variants. The bioinformatics of these variant alleles extended our knowledge of RhD variants, which was crucial for evaluating their impact to guide transfusion support and avoid immune-related blood transfusion reactions.
RhD 是最重要且最复杂的血型系统，因其具有高度多态性和免疫原性。RhD 变体可通过同种异体输血、器官移植和胎儿免疫引起免疫反应。由于各种等位基因形成的 RhD 变体的输血策略不同，因此广泛研究 RhD 变体的分子机制对于预防免疫相关的输血反应和胎儿免疫至关重要。
从中国浙江省的供体和患者中收集 RhD 变体。使用血清学法对表型进行分类。使用 PCR-SBT 方法分析基因的全长编码区。使用多重连接依赖性探针扩增 (MLPA) 检测分析基因型和基因拷贝数。使用 SWISS-MODLE 和 PyMOL 软件分析由变体等位基因引起的 RhD 的 3D 结构。使用 Polymorphism Phenotyping algorithm (PolyPhen-2)、Sorting Intolerant From Tolerant (SIFT) 和 Protein Variation Effect Analyzer (PROVEAN) 软件预测非同义取代的影响。
在所收集的 RhD 变体中，鉴定出 28 个不同的 变体等位基因，包括 3 个新的变体等位基因。RH-MLPA 检测对于确定基因的拷贝数是有利的。3D 同源建模预测蛋白质构象被破坏，这可能解释了 RhD 表位差异表达。总共确定了 14 个非同义突变对蛋白质结构有害。
我们揭示了中国东部 RhD 变体中存在的 等位基因的多样性。这些变体等位基因的生物信息学扩展了我们对 RhD 变体的认识，这对于评估它们的影响以指导输血支持和避免免疫相关的输血反应至关重要。
