Goldmacher V S, Lambert J M, Young A Y, Anderson J, Tinnel N L, Kornacki M, Ritz J, Blättler W A
J Immunol. 1986 Jan;136(1):320-5.
Expression of the common acute lymphoblastic leukemia antigen (CALLA) on the surface of individual cells of the human lymphoblastoid lines CW678, Namalwa, and Nalm-6, and the distribution of the antigen epitopes within the cell populations have been determined quantitatively with the murine monoclonal anti-CALLA antibody J5. The distribution of CALLA epitopes in the cell populations was analyzed by indirect immunofluorescence measured by using flow cytometry. The average number of CALLA epitopes per cell were measured by two assays: in a direct assay by binding 125I-labeled antibody J5 to cells, and indirectly by binding 125I-labeled protein A from Staphylococcus aureus to J5-coated cells. On average, CW678, Namalwa, and Nalm-6 cells bore about 1 X 10(4), 6 X 10(4), and 8 X 10(4) CALLA epitopes per cell respectively. Histograms of the absolute number of CALLA epitopes expressed by individual cells in the populations of CW678, Namalwa, and Nalm-6 cultures were generated by a combined analysis of all the binding data. This is the first example of histograms showing quantitative distribution of antigen epitopes. Previously, the expression of antigens by individual cells as obtained by flow cytometry was only presented in terms of relative fluorescence intensity of individual cells in cell populations.
利用鼠单克隆抗普通急性淋巴细胞白血病抗原(CALLA)抗体J5,已对人淋巴母细胞系CW678、Namalwa和Nalm-6单个细胞表面CALLA的表达以及抗原表位在细胞群体中的分布进行了定量测定。通过使用流式细胞术测定间接免疫荧光,分析了细胞群体中CALLA表位的分布。通过两种测定方法测量了每个细胞上CALLA表位的平均数量:一种直接测定方法是将125I标记的抗体J5与细胞结合,另一种间接方法是将来自金黄色葡萄球菌的125I标记蛋白A与包被有J5的细胞结合。平均而言,CW678、Namalwa和Nalm-6细胞每个细胞分别带有约1×10⁴、6×10⁴和8×10⁴个CALLA表位。通过对所有结合数据的综合分析,生成了CW678、Namalwa和Nalm-6培养物群体中单个细胞表达的CALLA表位绝对数量的直方图。这是显示抗原表位定量分布的直方图的首个实例。此前,通过流式细胞术获得的单个细胞的抗原表达仅以细胞群体中单个细胞的相对荧光强度来呈现。