Analysis of density and epitopes of D antigen on the surface of erythrocytes from DEL phenotypic individuals carrying the RHD1227A allele
Analysis of density and epitopes of D antigen on the surface of erythrocytes from DEL phenotypic individuals carrying the RHD1227A allele
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DOI：
10.2450/2013.0091-13
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发表时间：
2014-04-01
影响因子：
3.7
通讯作者：
Sun, Guang-Ming
中科院分区：
文献类型：
--
作者：
Gu, Juan;Sun, An-Yuan;Sun, Guang-Ming
背景。D抗原的特性是重要的，因为它们影响D变异细胞的免疫原性。一些关于正常D阳性、弱D和部分D病例抗原位点的研究已被报道，包括对高加索人DEL类型的综合分析。本研究的目的是评估中国人群中携带RHD1227A等位基因的亚洲型DEL表型个体红细胞表面的D抗原密度和表位。材料和方法。通过吸附、洗脱试验和序列特异性引物聚合酶链反应分析，在中国人群中共鉴定出154例携带RHD1227A等位基因的DEL表型个体。用流式细胞术检测这些个体红细胞表面的D抗原密度。用已知D抗原密度的红细胞样本作为标准。d -阴性和d -阳性个体的血液样本作为对照。此外，用18种针对不同D抗原表位的单克隆抗D抗体对携带RHD1227A等位基因的DEL个体红细胞表面的D抗原表位进行了研究。D阴性、D阳性和DEL个体红细胞膜表面D抗原荧光强度中位数均值分别为2.14 +/- 0.25、193.61 +/- 11.43和2.45 +/- 0.82。DEL样本估计每个细胞大约有22个D抗原。所有154例DEL患者的样本与18种不同D抗原表位特异性的单克隆抗D抗体反应阳性。在本研究中，携带RHD1227A等位基因的DEL个体红细胞表面的D抗原密度极低，每个细胞只有很少的抗原分子，但D抗原表位基本完整。
Background. The characteristics of the D antigen are important as they influence the immunogenicity of D variant cells. Several studies on antigenic sites have been reported in normal D positive, weak D and partial D cases, including a comprehensive analysis of DEL types in Caucasians. The aim of this study was to assess D antigen density and epitopes on the erythrocyte surface of Asian type DEL phenotypic individuals carrying the RHD1227A allele in the Chinese population.Materials and methods. A total of 154 DEL phenotypic individuals carrying the RHD1227A allele were identified through adsorption and elution tests and polymerase chain reaction analysis with sequence-specific primers in the Chinese population. D antigen density on the erythrocyte surface of these individuals was detected using a flow cytometric method. An erythrocyte sample with known D antigen density was used as a standard. Blood samples from D-negative and D-positive individuals were used as controls. In addition, D antigen epitopes on the erythrocyte surface of DEL individuals carrying the RHD1227A allele were investigated with 18 monoclonal anti-D antibodies specific for different D antigen epitopes.Results. The means of the median fluorescence intensity of D antigen on the erythrocyte membrane surface of D-negative, D-positive and DEL individuals were 2.14 +/- 0.25, 193.61 +/- 11.43 and 2.45 +/- 0.82, respectively. The DEL samples were estimated to have approximately 22 D antigens per cell. The samples from all 154 DEL individuals reacted positively with 18 monoclonal anti-D antibodies specific for different D antigen epitopes.Discussion. In this study, D antigen density on the erythrocyte surface of DEL individuals carrying the RHD1227A allele was extremely low, there being only very few antigenic molecules per cell, but the D antigen epitopes were grossly complete.