Gu Juan, Sun An-Yuan, Wang Xue-Dong, Shao Chao-Peng, Li Zheng, Huang Li-Hua, Pan Zhao-Lin, Wang Qing-Ping, Sun Guang-Ming
Department of Clinical Laboratory, The Fifth People's Hospital of Wuxi, Affiliated Hospital of Nanjing Medical University, Wuxi, Jiangsu, People's Republic of China.
Department of Clinical Laboratory, The Affiliated Provincial Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China.
Blood Transfus. 2014 Apr;12(2):244-9. doi: 10.2450/2013.0091-13. Epub 2013 Nov 21.
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.
背景:D抗原的特性很重要,因为它们会影响D变异细胞的免疫原性。关于正常D阳性、弱D和部分D病例的抗原位点已有多项研究报道,包括对白种人中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。 材料与方法:通过吸附洗脱试验和序列特异性引物聚合酶链反应分析,在中国人群中鉴定出154例携带RHD1227A等位基因的DEL表型个体。采用流式细胞术检测这些个体红细胞表面的D抗原密度。以已知D抗原密度的红细胞样本作为标准。将D阴性和D阳性个体的血样用作对照。此外,用18种针对不同D抗原表位的单克隆抗-D抗体研究携带RHD1227A等位基因的DEL个体红细胞表面的D抗原表位。 结果:D阴性、D阳性和DEL个体红细胞膜表面D抗原的中位荧光强度平均值分别为2.14±0.25、193.61±11.43和2.45±0.82。估计DEL样本每个细胞约有22个D抗原。所有154例DEL个体的样本与18种针对不同D抗原表位的单克隆抗-D抗体均呈阳性反应。 讨论: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,每个细胞仅有极少数抗原分子,但D抗原表位基本完整。
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