Rumeu M T, Suárez M A, Morales S, Rotger R
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense, Madrid, Spain.
J Appl Microbiol. 1997 Jan;82(1):19-31. doi: 10.1111/j.1365-2672.1997.tb03293.x.
Seventy-six Salmonella enteritidis, three Salmonella virchow and one Salmonella bradenrup strains were screened for enterotoxigenicity by using the Chinese hamster ovary (CHO), Y1 adrenal, Vero and HeLa cell tests. All the strains gave positive reactions for enterotoxin production, except one, and the relative sensitivity to the toxin exhibited by the different cell lines was evaluated. An enterotoxic activity has been identified in sonicated extracts of Salm. enteritidis. This enterotoxin was purified on Agarose A-5m (Bio-Rad) and Superose 12 HR 10/30 column. The enterotoxic activity was eluted from the Superose column in the first peak. Like Vibrio cholerae toxin CT and Escherichia coli enterotoxin LT, it was blocked by GM1 ganglioside, but at a higher concentration. In addition, a cytotoxic factor has been partially identified. The procedure for isolating the cytotoxin included ammonium sulphate precipitation, size-exclusion chromatography and anion exchange chromatography. This cytotoxin factor caused inhibition of protein synthesis in cultured cells, as determined by flow cytometry and [3H]-leucine incorporation. Flow cytometry analysis also showed an activation of CHO cells when exposed to this cytotoxic factor resulting in a state of active growth. Cytotoxic activity was not blocked by gangliosides.
采用中国仓鼠卵巢(CHO)细胞、Y1肾上腺细胞、Vero细胞和HeLa细胞试验,对76株肠炎沙门氏菌、3株维尔肖沙门氏菌和1株布拉登鲁普沙门氏菌进行了肠毒素产生能力的筛选。除1株外,所有菌株的肠毒素产生试验均呈阳性反应,并评估了不同细胞系对该毒素的相对敏感性。已在肠炎沙门氏菌的超声提取物中鉴定出一种肠毒素活性。该肠毒素在琼脂糖A-5m(伯乐公司)和Superose 12 HR 10/30柱上进行纯化。肠毒素活性在Superose柱上的第一个峰中被洗脱出来。与霍乱弧菌毒素CT和大肠杆菌肠毒素LT一样,它被GM1神经节苷脂阻断,但所需浓度更高。此外,还部分鉴定出一种细胞毒素因子。分离细胞毒素的步骤包括硫酸铵沉淀、尺寸排阻色谱和阴离子交换色谱。通过流式细胞术和[3H]-亮氨酸掺入法测定,这种细胞毒素因子可抑制培养细胞中的蛋白质合成。流式细胞术分析还显示,当CHO细胞暴露于这种细胞毒素因子时会被激活,从而进入活跃生长状态。细胞毒素活性不受神经节苷脂的阻断。