Hudler H, Stemberger H, Scheiner O, Kollaritsch H, Wiedermann G
Tropenmed Parasitol. 1983 Dec;34(4):248-52.
Cytotoxic action against K562-tissue culture cells was investigated under various conditions with a Chromium-release-assay. When amoebae and target cells were centrifuged together, pathogenic strains of amoebae induced a very fast increase of target cell lysis (up to 50% of maximum lysis after 10 minutes). Only a minor degree of target cell lysis resulted, however, when amoebae and K562 cells were kept in suspension. When amoebae were eliminated selectively by addition of complement 10 minutes after starting the experiment, this fast increase of lysis could not be prevented. These observations suggest that the cytotoxic action might take place in two distinct phases. The first step ("lethal hit") seems to be temperature-independent, whereas a temperature of 37 degrees C is necessary for the second step to occur during which cytoplasmic material is released (chromium release). The presence of amoebae is not necessary for the second step. When amoebae together with and target cells are kept in suspension, amoebae lost their capability of setting the "lethal hit" with increasing time of coincubation. It seems, as if the "lethal hit" cannot be accomplished effectively under the conditions of suspension: cytotoxic substances released by the amoebae cannot be transferred to the target cells and are lost in the fluid phase. Thereby, the amoebae are depleted of such substances. Thus, a stable contact between amoebae and target cells for at least a few minutes seems to be necessary for the expression of cytotoxicity.
采用铬释放试验,在不同条件下研究了对K562组织培养细胞的细胞毒性作用。当变形虫与靶细胞一起离心时,变形虫的致病菌株会导致靶细胞裂解迅速增加(10分钟后达到最大裂解的50%)。然而,当变形虫和K562细胞保持悬浮状态时,仅导致轻微程度的靶细胞裂解。在实验开始10分钟后通过添加补体选择性地清除变形虫时,这种裂解的快速增加无法得到阻止。这些观察结果表明,细胞毒性作用可能分两个不同阶段发生。第一步(“致死打击”)似乎与温度无关,而第二步发生时需要37摄氏度的温度,在此期间细胞质物质被释放(铬释放)。第二步发生时不需要变形虫的存在。当变形虫与靶细胞一起保持悬浮状态时,随着共孵育时间的增加,变形虫失去了进行“致死打击”的能力。似乎在悬浮条件下“致死打击”无法有效完成:变形虫释放的细胞毒性物质无法转移到靶细胞并在液相中丢失。由此,变形虫消耗了此类物质。因此,变形虫与靶细胞之间至少稳定接触几分钟似乎是细胞毒性表达所必需的。