A HIGHLY SENSITIVE CELL-LINE, WEHI-164 CLONE 13, FOR MEASURING CYTOTOXIC FACTOR TUMOR-NECROSIS-FACTOR FROM HUMAN-MONOCYTES
A HIGHLY SENSITIVE CELL-LINE, WEHI-164 CLONE 13, FOR MEASURING CYTOTOXIC FACTOR TUMOR-NECROSIS-FACTOR FROM HUMAN-MONOCYTES
复制标题
DOI：
10.1016/0022-1759(86)90322-4
复制
发表时间：
1986-01-01
影响因子：
2.2
通讯作者：
NISSENMEYER, J
中科院分区：
文献类型：
--
作者：
ESPEVIK, T;NISSENMEYER, J
通过有限稀释 WEHI 164 小鼠纤维肉瘤细胞，我们分离出了细胞系。 WEHI 164 克隆 13，对源自人单核细胞的细胞毒性因子 (CF) 极其敏感。通过在MTT四唑细胞毒性测定中使用WEHI 164克隆13，发现来自活化单核细胞的CF上清液必须稀释105-106倍才能达到产生50%死细胞的剂量（LD50）。通过比较不同靶细胞的LD50。发现 WEHI 164 克隆 13 细胞对 CF 诱导的细胞毒性的敏感性比 WEHI 164 亲代细胞高约 103 倍，与放线菌素 D 处理的 L929 细胞相比约高 102 倍。用放线菌素 D 处理 WEHI 164 克隆 13 细胞并没有增加它们对 CF 诱导的细胞毒性的敏感性。重组肿瘤坏死因子(rTNF)还介导对WEHI 164克隆13细胞的高细胞毒性，LD50为2倍。 10-3 纳克/毫升。中和CF抗血清完全抑制了rTNF的毒性活性。 WEHI 164 克隆 13 细胞对单核细胞介导的细胞毒性高度敏感，因为 1-2 个单核细胞能够杀死至少 5000 个这些靶细胞。中和TNF抗血清完全抑制单核细胞介导的细胞毒性。这些结果表明CF上清液和单核细胞对WEHI 164克隆13细胞介导的高水平细胞毒性是由于TNF作为效应分子。
By limiting dilution of WEHI 164 mouse fibrosarcoma cells we have isolated a cell line. WEHI 164 clone 13, which is extremely sensitive to cytotoxic factor (CF) derived from human monocytes. By using WEHI 164 clone 13 in a MTT tetrazolium cytotoxicity assay it was found that CF supernatants from activated monocytes had to be diluted 105-106 times to reach the dose which produced 50% dead cells (LD50). By comparing the LD50 of different target cells. WEHI 164 clone 13 cells were found to be approximately 103 times more sensitive for CF-induced cytotoxicity as compared to WEHI 164 parental cells and approximately 102 times more sensitive as compared to actinomycin D-treated L929 cells. Treatment of the WEHI 164 clone 13 cells weith actinomycin D did not increase their sensitivity for CF-induced cytotoxicity. Recombinant tumor necrosis factor (rTNF) also mediated high cytotoxicity towards WEHI 164 clone 13 cells, with an LD50 of 2 .times. 10-3 ng/ml. Neutralizing CF antiserum completely inhibited the toxic activity of rTNF. WEHI 164 clone 13 cells were highly sensitive to monocyte-mediated cytotoxicity in that 1-2 monocytes were able to kill at least 5000 of these target cells. Neutralizing TNF antiserum completely inhibited monocyte-mediated cytotoxicity. These results indicate that the high level of cytotoxicity mediated by CF supernatants and monocytes on WEHI 164 clone 13 cells is due to TNF as the effector molecule.