使用非贴壁细胞FM3A对产气荚膜梭菌粗制肠毒素细胞毒性活性进行流式细胞术检测。

Flow cytometric assay for cytotoxic activity of crude Clostridium perfringens enterotoxin using non-adherent cell FM3A.

作者信息

Hu D, Sugii S, Kusunoki H, Uemura T

机构信息

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, College of Agriculture, Osaka Prefecture University, Sakai, Japan.

出版信息

FEMS Immunol Med Microbiol. 1995 Dec;12(3-4):239-44. doi: 10.1111/j.1574-695X.1995.tb00198.x.

DOI:10.1111/j.1574-695X.1995.tb00198.x

PMID:8745009

Abstract

The flow cytometric assay method was tested for the cytotoxic activity of Clostridium perfringens enterotoxin (CPE) in culture using mouse mammary carcinoma cell line FM3A stained with propidium iodide (PI). From the results obtained, FM3A cells proved to be susceptible to CPE. A reproducible dose-response curve with FM3A was obtained between crude CPE at 13.9-109 ng/ml and between purified CPE at 40-400 ng/ml, respectively. These findings indicate that non-adherent FM3A is preferable to determine the cytotoxic activity of CPE because it can be used without detachment procedures with trypsinin compared with adherent African monkey kidney cell line (Vero cells). Furthermore, the flow cytometry with non-adherent cell FM3A stained with PI only proved to be a useful method to determine the biological activity of CPE in culture isolates.

摘要

采用碘化丙啶（PI）染色的小鼠乳腺癌细胞系FM3A，通过流式细胞术检测产气荚膜梭菌肠毒素（CPE）在培养物中的细胞毒性活性。从获得的结果来看，FM3A细胞被证明对CPE敏感。分别在13.9 - 109 ng/ml的粗制CPE和40 - 400 ng/ml的纯化CPE之间获得了与FM3A相关的可重复剂量反应曲线。这些发现表明，非贴壁的FM3A更适合用于确定CPE的细胞毒性活性，因为与贴壁的非洲猴肾细胞系（Vero细胞）相比，它无需用胰蛋白酶进行分离程序即可使用。此外，仅用PI染色的非贴壁细胞FM3A进行流式细胞术被证明是一种确定培养物分离物中CPE生物活性的有用方法。