Radosević K, de Grooth B G, Greve J
University of Twente, Department of Applied Physics, Enschede, The Netherlands.
Cytometry. 1993;14(5):535-40. doi: 10.1002/cyto.990140513.
A flow cytometric method for the simultaneous quantification and immunophenotyping of conjugates formed by human peripheral blood lymphocytes (PBL) and K562 cells has been developed. The method uses three fluorescent probes. One of the fluorescent probes (F-18) is used for labeling of PBL prior to incubation with K562 cells. After incubation the cells are treated with monoclonal antibodies labeled with phycoerythrin and Red613, respectively. The combination of F-18 fluorescence and light scattering signals enables identification and quantification of the conjugates while the fluorescence of the monoclonal antibodies provides information about the phenotype of the conjugate forming cells. Results obtained using different monoclonal antibodies are presented. The highest conjugate forming capacity has been found in the CD56+CD8+ population while the CD4+CD8- population has shown the lowest capacity to form conjugates. The influence of a washing step on the conjugate formation is discussed. The possibility to use the method in combination with a cytotoxicity assay is indicated.
已开发出一种流式细胞术方法，用于同时定量分析人外周血淋巴细胞（PBL）与K562细胞形成的结合物并进行免疫表型分析。该方法使用三种荧光探针。其中一种荧光探针（F-18）用于在与K562细胞孵育前标记PBL。孵育后，细胞分别用藻红蛋白和Red613标记的单克隆抗体处理。F-18荧光和光散射信号的组合能够鉴定和定量结合物，而单克隆抗体的荧光则提供有关形成结合物细胞表型的信息。展示了使用不同单克隆抗体获得的结果。发现CD56 + CD8 +群体形成结合物的能力最高，而CD4 + CD8-群体形成结合物的能力最低。讨论了洗涤步骤对结合物形成的影响。指出了将该方法与细胞毒性测定结合使用的可能性。
