Heeg K, Reimann J, Kabelitz D, Hardt C, Wagner H
J Immunol Methods. 1985 Mar 18;77(2):237-46. doi: 10.1016/0022-1759(85)90036-5.
Interleukin 2 (IL-2) activity is tested in conditioned media by assessing its ability to support proliferation of selected IL-2 dependent T cell lines, conventionally measured by [3H]thymidine incorporation. Here, we compare this [3H]thymidine uptake test for measuring IL-2 activity with a rapid and sensitive colorimetric method which is based on the ability of viable cells to cleave 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). The sensitivity of the colorimetric method was dependent on the indicator cell line used, being greatest with the cytotoxic T cell line 16 (CTLL-16). The colorimetric method is at least as sensitive as [3H]thymidine uptake tests, does not rely on radioactivity, and is ideally suited to screen large numbers of individual samples for IL-2 activity. The latter point was demonstrated by calculating IL-2-producing helper T cell frequencies in heterogeneous murine lymphocyte populations: in this assay, splenic T cells were clonally expanded under limiting dilution conditions and supernatants conditioned by these in vitro growing T cell clones were tested for IL-2 activity with the colorimetric method. This allowed us to obtain reliable estimates of the frequency of progenitor cells of IL-2-producing T cell clones in various populations.
通过评估其支持选定的白细胞介素2（IL-2）依赖性T细胞系增殖的能力，在条件培养基中检测IL-2活性，传统上通过[3H]胸苷掺入来测量。在此，我们将这种用于测量IL-2活性的[3H]胸苷摄取试验与一种基于活细胞裂解3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑（MTT）能力的快速灵敏比色法进行比较。比色法的灵敏度取决于所使用的指示细胞系，对细胞毒性T细胞系16（CTLL-16）最为敏感。比色法至少与[3H]胸苷摄取试验一样灵敏，不依赖放射性，非常适合筛选大量单个样本的IL-2活性。通过计算异基因小鼠淋巴细胞群体中产生IL-2的辅助性T细胞频率证明了后一点：在该试验中，脾T细胞在有限稀释条件下进行克隆扩增，并用比色法检测这些体外生长的T细胞克隆产生的上清液中的IL-2活性。这使我们能够可靠地估计不同群体中产生IL-2的T细胞克隆祖细胞的频率。
