Zeevi A, Chiu K M, Duquesnoy R J
J Immunol. 1980 Sep;125(3):1130-5.
This report describes a method of growing soft agar colonies of human T lymphocytes activated in the MLR. Two types of colonies were demonstrated: lower colonies grew within the agar layer, and upper colonies grew on the surface of the agar layer. Three days of priming the lymphocytes in the MLR and the use of supernatants of day-3 MLR cultures to provide T cell colony growth factor were necessary for optimal colony formation. Lymphocytes obtained from colonies were grown in long-term (2 to 4 weeks) cultures to generate sufficient numbers of cells to be tested in different functional assays. Cells from both types of colonies exhibited PLT activity. Upper colony cells showed considerably higher CML activity than lower colony cells (mean percent cytotoxicity 37 +/- 5 vs 6 +/- 3). Cells from both types of colonies contained radiosensitive suppressor cell activity that inhibited the primary MLR. The suppressor cell effect of lower colony cells was specific for the original stimulator, but upper colony cells displayed nonspecific suppressive effects. For both types of colony cells, it appeared that suppressive effects were unrelated to the CML activity of these cells. These data suggest that the soft agar colony assay offers a promising approach to separate subpopulations of lymphocytes activated in the MLR.
本报告描述了一种培养在混合淋巴细胞反应（MLR）中活化的人T淋巴细胞软琼脂集落的方法。展示了两种类型的集落：下层集落在琼脂层内生长，上层集落在琼脂层表面生长。在MLR中对淋巴细胞进行3天的预刺激，并使用第3天MLR培养物的上清液来提供T细胞集落生长因子，对于最佳集落形成是必要的。从集落中获得的淋巴细胞在长期（2至4周）培养物中生长，以产生足够数量的细胞用于不同的功能测定。两种类型集落的细胞均表现出PLT活性。上层集落细胞显示出比下层集落细胞高得多的CML活性（平均细胞毒性百分比为37±5对6±3）。两种类型集落的细胞都含有抑制原发性MLR的放射敏感性抑制细胞活性。下层集落细胞的抑制细胞效应对原始刺激物具有特异性，但上层集落细胞表现出非特异性抑制作用。对于两种类型的集落细胞，抑制作用似乎与这些细胞的CML活性无关。这些数据表明，软琼脂集落测定法为分离在MLR中活化的淋巴细胞亚群提供了一种有前景的方法。
