Flow cytometric characterization of normal and variant cells with monoclonal antibodies specific for glycophorin A.
Flow cytometric characterization of normal and variant cells with monoclonal antibodies specific for glycophorin A.
复制标题
使用血型糖蛋白 A 特异性单克隆抗体对正常细胞和变异细胞进行流式细胞术表征。
DOI：
10.4049/jimmunol.134.6.4009
复制
发表时间：
1985
影响因子：
4.4
通讯作者：
R. Jensen
中科院分区：
文献类型：
--
作者：
R. Langlois;W. Bigbee;R. Jensen
定量免疫荧光测量红细胞标记单克隆抗体糖蛋白A (GPA)，以评估这些抗体与正常和变异细胞类型的结合水平。本研究中使用的7种抗体包括两种优先与M型GPA结合的抗体，三种优先与N型GPA结合的抗体，还有两种与这两种抗体都结合得很好。流式细胞术分析显示，大多数抗体的结合特异性大于100倍，并且显示三种抗体与细胞结合的GPA的亲和力约为10(9)M（-1）。这些数据还表明，GPA的表达水平在细胞和个体之间的差异小于10%。用以下不同形式的糖蛋白对人红细胞进行流量测量：Mc、Mg、Mk、En(F)、En（UK）、Mi-I、Mi-II、Mi-III、S-s-U-、Tn+和St（a+）。其他被分析的细胞类型包括来自黑猩猩、恒河猴、非洲绿猴和卷尾猴的红细胞，以及来自人类红细胞白血病细胞系K562的细胞。我们的7种抗体的流动分析显示，这些细胞类型具有独特的标记模式，与已知或推断的糖蛋白改变一致。在大多数情况下，变异等位基因表达在正常水平。我们的研究结果支持了其他的观察结果，即变异En（UK）和St（a+）含有GPA-GPB杂交蛋白，并表明它们的表达水平在很大程度上取决于杂交基因的3'端。
Quantitative immunofluorescence measurements were performed on erythrocytes labeled with monoclonal antibodies to glycophorin A (GPA) to assess the level of binding of these antibodies to normal and variant cell types. The seven antibodies used in this study include two that bind preferentially to the M form of GPA, three that bind preferentially to the N form, and two that bind equally well to both. Flow cytometric analysis of mixtures of cells differing in M,N type showed binding specificities of greater than 100-fold for most of the antibodies, and showed that three antibodies bind cell-bound GPA with an affinity of approximately 10(9) M(-1). These data also showed that the level of expression of GPA varies by less than 10% from cell to cell and from individual to individual. Flow measurements were also done on human erythrocytes with the following variant forms of glycophorin: Mc, Mg, Mk, En(F), En(UK), Mi-I, Mi-II, Mi-III, S-s-U-, Tn+, and St(a+). Other cell types analyzed included erythrocytes from chimpanzee, rhesus, African green, and capuchin monkeys, and cells from the human erythroleukemia cell line, K562. Flow analysis with our seven antibodies showed these cell types have distinctive labeling patterns consistent with the known or inferred altered glycophorins presented on these cells. In most cases, variant alleles were expressed at normal levels. Our results support other observations that the variants En(UK) and St(a+) contain hybrid GPA-GPB proteins, and suggest that their level of expression is largely determined by the 3' end of the hybrid genes.