Kahle P, Wernet P, Rehbein A, Kumbier I, Pawelec G
Scand J Immunol. 1981 Nov;14(5):493-502.
Conditions for the cloning of alloactivated human lymphocytes by limiting dilution in the presence of interleukin 2 (IL 2) and filler cells have been investigated. Cloning efficiencies were extremely high (at least 40%) when IL 2 derived from pooled peripheral blood mononuclear cells (PBM), stimulated with phytohaemagglutinin in the presence of irradiated Epstein-Barr virus-transformed lymphoid cell lines (LCL), was used in combination with irradiated pooled PBM as filler cells. Cloning efficiencies were reduced almost twofold using autologous filler cells and were further reduced dramatically using autologous IL2, although some clones could still be obtained. In addition, cloning efficiencies were acceptable (over 30%) when IL 2 produced spontaneously from the leukaemic cell Jurkat (M-N) was used. In contrast, sheep erythrocytes and LCL as filler cells failed to allow successful cloning, although LCL were beneficial in the expansion of cloned populations. Other factors being equal, the use of pooled PBM as filler cells and highly active preparations of IL 2 will be the method of choice for cloning functional human T lymphocytes.
研究了在白细胞介素2（IL-2）和填充细胞存在的情况下，通过有限稀释法克隆同种异体激活的人淋巴细胞的条件。当使用来自汇集的外周血单个核细胞（PBM）的IL-2（在经辐照的爱泼斯坦-巴尔病毒转化的淋巴母细胞系（LCL）存在下用植物血凝素刺激）与经辐照的汇集PBM作为填充细胞联合使用时，克隆效率极高（至少40%）。使用自体填充细胞时克隆效率降低近两倍，使用自体IL-2时克隆效率进一步大幅降低，尽管仍可获得一些克隆。此外，当使用白血病细胞Jurkat（M-N）自发产生的IL-2时，克隆效率是可以接受的（超过30%）。相比之下，绵羊红细胞和LCL作为填充细胞无法成功克隆，尽管LCL对克隆群体的扩增有益。在其他因素相同的情况下，使用汇集的PBM作为填充细胞和高活性的IL-2制剂将是克隆功能性人T淋巴细胞的首选方法。
