Laboratory of Molecular Biology Graduate School of Environmental Health Sciences, Azabu University, Fuchinobe, Chuo-ku Sagamihara, Japan.
J Basic Microbiol. 2012 Oct;52(5):559-65. doi: 10.1002/jobm.201100297. Epub 2012 Feb 23.
We aimed to clarify if Campylobacter lari exerts a cytolethal distending toxin (CDT) effect on HeLa cells. Campylobacter cell lysates (CCLys) from C. jejuni 81-176 and urease-positive thermophilic Campylobacter (UPTC) CF89-12 and UPTC NCTC12893 isolates were shown to exert a CDT effect on HeLa cells with morphological changes examined by Giemsa staining and microscopy. However, Campylobacter lari JCM2530(T) isolate showed no effect. In addition, Campylobacter cell culture supernatant wash gave low or absent toxic effects with both C. jejuni and C. lari organisms. When western blot analysis was carried out to clarify if there was a CDTB effect in the CCLys and soluble fractions from Campylobacter isolates, which had a CDT effect on HeLa cells or did not have any effect, anti-recombinant CjCDTB antibodies identified an immunoreactively positive signal at around approximately 25 kDa on all the C. lari isolates examined, as well as the C. jejuni 81116 strain. Thus, all the Campylobacter isolates including those without any CDT effect were shown to express CDTB at the translational level.
我们旨在阐明空肠弯曲菌是否对 HeLa 细胞产生细胞致死性扩张毒素 (CDT) 效应。从空肠弯曲菌 81-176 和产脲酶嗜热弯曲菌 (UPTC) CF89-12 和 NCTC12893 分离株的弯曲菌细胞裂解物 (CCLys) 显示对 HeLa 细胞产生 CDT 效应,通过吉姆萨染色和显微镜检查观察到形态变化。然而,空肠弯曲菌 JCM2530(T) 分离株没有这种作用。此外,用空肠弯曲菌和空肠弯曲菌进行细胞培养上清液洗涤时,无论是否存在 CDT 效应,都表现出低毒性或无毒性。当进行 Western blot 分析以阐明对 HeLa 细胞具有 CDT 效应或没有任何效应的弯曲菌分离株的 CCLys 和可溶性部分是否存在 CDTB 效应时,抗重组 CjCDTB 抗体在所有检测到的空肠弯曲菌分离株以及空肠弯曲菌 81116 株上均在大约 25 kDa 左右识别出免疫反应性阳性信号。因此,所有包括没有任何 CDT 效应的弯曲菌分离株都在翻译水平上显示出 CDTB 的表达。