Voogt P J, Fibbe W E, Veenhof W F, Brand A, Goulmy E, van Rood J J, Falkenburg J H
Laboratory of Experimental Hematology, University Medical Center, Leiden, The Netherlands.
Leukemia. 1987 May;1(5):427-31.
Several techniques are available for the serological analysis of antigenic determinants on human hematopoietic progenitor cells (HPC). However, techniques for the recognition of cellularly defined antigens on such progenitor cells have not yet been described. We therefore developed an in vitro cellular cytotoxicity assay, with bone marrow cells as target cells. In this assay specific cytotoxic T lymphocyte (CTL) lines are used as effectors for cell-mediated cytolysis of bone marrow mononuclear cells that express the antigens for which the CTLs were primed in a mixed lymphocyte culture. As a model we used CTL lines against HLA-A2 or -B7 determinants. By using effector-target ratios varying from 1:2 to 4:1, 4 hr of incubation of these CTL lines with bone marrow mononuclear cells from HLA-A2 or -B7 positive donors resulted in a specific dose-dependent growth inhibition up to 100% of myeloid (CFU-GM), erythroid (BFU-E), and multipotential (CFU-GEMM) HPC. In contrast no inhibition of HPC was observed using mononuclear bone marrow cells from HLA-A2 or -B7 negative individuals as target cells. Experiments in which cell-cell contact was prevented showed that the antigen-specific lysis of HPC was dependent on intimate cell-cell contact between effector-CTLs and bone marrow target cells. Our results show that this cell-mediated cytotoxicity assay can be used as a sensitive and specific tool for the study of cellularly defined antigens on human hematopoietic progenitor cells.
有几种技术可用于对人类造血祖细胞（HPC）上的抗原决定簇进行血清学分析。然而，尚未描述用于识别此类祖细胞上细胞定义抗原的技术。因此，我们开发了一种以骨髓细胞为靶细胞的体外细胞毒性测定法。在该测定法中，特异性细胞毒性T淋巴细胞（CTL）系用作效应细胞，用于对表达抗原的骨髓单个核细胞进行细胞介导的细胞溶解，这些抗原是CTL在混合淋巴细胞培养中被致敏的对象。作为模型，我们使用了针对HLA - A2或 - B7决定簇的CTL系。通过使用从1:2到4:1变化的效应细胞与靶细胞比例，将这些CTL系与来自HLA - A2或 - B7阳性供体的骨髓单个核细胞孵育4小时，导致髓系（CFU - GM）、红系（BFU - E）和多能（CFU - GEMM）HPC出现特异性剂量依赖性生长抑制，最高可达100%。相比之下，使用来自HLA - A2或 - B7阴性个体的骨髓单个核细胞作为靶细胞时，未观察到HPC受到抑制。防止细胞 - 细胞接触的实验表明，HPC的抗原特异性裂解依赖于效应CTL与骨髓靶细胞之间紧密的细胞 - 细胞接触。我们的结果表明，这种细胞介导的细胞毒性测定法可作为研究人类造血祖细胞上细胞定义抗原的灵敏且特异的工具。
