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首页> 外文期刊>Journal of Clinical Microbiology >Effects of cycloheximide and puromycin on cytotoxic activity of Escherichia coli verocytotoxin (Shiga-like toxin).
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Effects of cycloheximide and puromycin on cytotoxic activity of Escherichia coli verocytotoxin (Shiga-like toxin).

机译：环己酰亚胺和嘌呤霉素对大肠杆菌Verocytotoxin（志贺样毒素）的细胞毒活性的影响。

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Verocytotoxin (VT)-producing Escherichia coli is closely associated with hemorrhagic colitis and hemolytic uremic syndrome. The diagnosis of this infection requires the demonstration of VT activity in fecal filtrates or the isolation of VT-producing E. coli from stools. To improve the sensitivity of the Vero cell assay for detecting VT, we investigated the interaction between this toxin and cycloheximide and puromycin, agents which, like VT and the related Shiga toxin, are protein synthesis inhibitors. Cycloheximide-treated cells were found to be about eightfold more sensitive to VT, this effect being most pronounced when the drug was added before the toxin. In contrast, puromycin treatment had an antagonistic effect in that it decreased the sensitivity of the cells to VT. In assays of VT in fecal filtrates, the addition of cycloheximide (at 4 to 8 micrograms/ml) increased the sensitivity without affecting the specificity of the assays. Likewise, the use of cycloheximide led to an increase in the sensitivity of the serum VT-neutralizing antibody test by a factor of over eightfold.
机译：产生毒素的大肠埃希菌与出血性结肠炎和溶血性尿毒症综合征密切相关。诊断这种感染需要在粪便滤液中证明VT活性或从粪便中分离出产生VT的大肠杆菌。为了提高Vero细胞测定法检测VT的敏感性，我们研究了该毒素与环己酰亚胺和嘌呤霉素之间的相互作用，这些试剂与VT和相关的志贺毒素一样，都是蛋白质合成抑制剂。发现用环己二酰亚胺处理的细胞对VT的敏感性高大约八倍，当在毒素之前添加药物时，这种作用最为明显。相反，嘌呤霉素处理具有拮抗作用，因为它降低了细胞对VT的敏感性。在粪便滤液中进行VT分析时，添加环己酰亚胺（4至8微克/毫升）可提高灵敏度，而不会影响分析的特异性。同样，使用环己酰亚胺导致血清VT中和抗体测试的敏感性增加了八倍以上。

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