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首页> 外文期刊>Infection and immunity >Effects of Staphylococcal Alpha-, Beta-, Delta-, and Gamma-Hemolysins on Human Diploid Fibroblasts and HeLa Cells: Evaluation of a New Quantitative Assay for Measuring Cell Damage
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Effects of Staphylococcal Alpha-, Beta-, Delta-, and Gamma-Hemolysins on Human Diploid Fibroblasts and HeLa Cells: Evaluation of a New Quantitative Assay for Measuring Cell Damage

机译：葡萄球菌的α-，β-，δ-和γ-溶血素对人二倍体成纤维细胞和HeLa细胞的影响：一种新的定量测定细胞损伤的定量测定方法的评估

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Human diploid embryonic lung fibroblasts and HeLa cells were cultivated in Eagle minima? essential medium supplemented with 10% calf serum. Monolayer cultures were labeled with 3H-uridine and treated with highly purified staphylococcal alpha-, beta-, delta-, or gamma-hemolysin. The release of soluble radioactive substances into the medium was used as an indicator of damage to the cell membrane after treatment with each hemolysin. The assay method described is simple, sensitive, and rapid. It allows quantitative estimation of changes in membrane permeability to be detected before a morphological damage is observed microscopically. Upon incubation for up to 30 min with highly purified staphylococcal hemolysins, only delta-hemolysin caused release of a significant amount of tritiated substances from fibroblasts. Such leakage occurred immediately after addition of delta-lysin and was independent of temperature. With minor exceptions, this was similar to the release of isotopes after treatment of the cells with the nonionic detergent Triton X-100. Treatment of fibroblasts with combinations of two or three of these toxins gave neither a synergistic nor an antagonistic effect. Evidence is presented which indicates that delta-hemolysin is the only important fibroblast damaging activity in crude preparations of extracellular proteins of four strains of S. aureus, whereas HeLa cells are susceptible also to purified alpha-toxin.
机译：人类二倍体胚胎肺成纤维细胞和HeLa细胞在Eagle minima中培养。必需培养基，补充10％小牛血清。用 3 H-尿苷标记单层培养物，并用高度纯化的葡萄球菌α-，β-，δ-或γ-溶血素处理。用每种溶血素处理后，可溶性放射性物质向培养基中的释放被用作细胞膜损伤的指标。所描述的测定方法简单，灵敏且快速。它允许在显微镜观察到形态损伤之前定量检测膜渗透性的变化。与高度纯化的葡萄球菌溶血素孵育30分钟后，仅δ-溶血素会从成纤维细胞中释放出大量tri化物质。这种泄漏是在加入δ-溶素后立即发生的，并且与温度无关。除少数例外，这类似于用非离子型清洁剂Triton X-100处理细胞后同位素的释放。用两种或三种这些毒素的组合处理成纤维细胞既没有协同作用也没有拮抗作用。提出的证据表明，δ-溶血素是四种菌株 S的细胞外蛋白粗制品中唯一重要的成纤维细胞破坏活性。金黄色葡萄球菌，而HeLa细胞也容易感染纯化的α-毒素。

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