Nathan C F, Brukner L H, Silverstein S C, Cohn Z A
J Exp Med. 1979 Jan 1;149(1):84-99. doi: 10.1084/jem.149.1.84.
Lymphoma cells were rapidly lysed by activated macrophages and granulocytes in the presence of PMA. Release of 51Cr from lymphoma cells correlated closely with their destruction as viewed by scanning electron microscopy, and with reduction in the number of trypan blue-excluding cells. The standard assay involved 51 Cr release measured at 4.5 h, but injury appeared to be complete in 1 h. Of eight different types of effector cells tested, only those releasing abundant H2O2 in response to PMA were effective, that, is BCG-, C. parvum-, or casein-activated macrophages, or thioglycollate-elicited granulocytes. Normal macrophages, J774 cells, or macrophages elicited with thioglycollate broth or proteose-peptone were ineffective. BCG-activated macrophages and granulocytes caused 50% specific release of 51Cr from P388 lymphoma cells at E:T ratios between 1.4 and 4.5, and from mouse erythrocytes at E:T ratios of 0.017 to 0.025. 10 types of target cells varied widely in their susceptibility to lysis by reagent H2O2, with one-half maximal lysis occurring at H2O2 concentrations ranging from 3.63 X 10(-6) M to 3.85 X 10(-5) M. Effector cells were expected to generate approximately that much H2O2 during the period of injury. Susceptibility of the target cells to lysis by PMA-triggered granulocytes correlated closely with their sensitivity to H2O2 (r = 0.98). The membrane-active agents LPS and digitonin, which did not trigger H2O2 release, did not trigger cytotoxicity. The dose-response curve for triggering of H2O2 release by PMA was identical to that for triggering cytotoxicity. These results provided strong circumstantial evidence for the importance of H2O2 in extracellular cytolysis by activated macrophages and granulocytes when pharmacologically triggered.
在佛波酯（PMA）存在的情况下，淋巴瘤细胞会被活化的巨噬细胞和粒细胞迅速裂解。从淋巴瘤细胞中释放的51Cr与通过扫描电子显微镜观察到的细胞破坏密切相关，也与台盼蓝拒染细胞数量的减少相关。标准检测方法是在4.5小时时测量51Cr的释放，但损伤似乎在1小时内就已完成。在测试的八种不同类型的效应细胞中，只有那些在PMA刺激下释放大量过氧化氢（H2O2）的细胞才有效，即卡介苗（BCG）、微小隐孢子虫或酪蛋白活化的巨噬细胞，或巯基乙酸盐诱导的粒细胞。正常巨噬细胞、J774细胞，或用巯基乙酸盐肉汤或蛋白胨诱导的巨噬细胞无效。BCG活化的巨噬细胞和粒细胞在效应细胞与靶细胞（E:T）比例为1.4至4.5时，能使P388淋巴瘤细胞特异性释放50%的51Cr，在E:T比例为0.017至0.025时，能使小鼠红细胞特异性释放50%的51Cr。10种靶细胞对试剂H2O2裂解的敏感性差异很大，半数最大裂解发生在H2O2浓度范围为3.63×10(-6) M至3.85×10(-5) M时。预计效应细胞在损伤期间会产生大约那么多的H2O2。靶细胞对PMA触发的粒细胞裂解的敏感性与其对H2O2的敏感性密切相关（r = 0.98）。不触发H2O2释放的膜活性剂脂多糖（LPS）和洋地黄皂苷不会触发细胞毒性。PMA触发H2O2释放的剂量反应曲线与触发细胞毒性的曲线相同。这些结果提供了有力的间接证据，证明在药理学触发下，H2O2在活化的巨噬细胞和粒细胞的细胞外细胞溶解中具有重要作用。
