Petit L, Gibert M, Gillet D, Laurent-Winter C, Boquet P, Popoff M R
Unité des Toxines Microbiennes, Institut Pasteur, Paris, France.
J Bacteriol. 1997 Oct;179(20):6480-7. doi: 10.1128/jb.179.20.6480-6487.1997.
Epsilon-toxin is produced by Clostridium perfringens types B and D and is responsible for a rapidly fatal enterotoxemia in animals, which is characterized by edema in several organs due to an increase in blood vessel permeability. The Madin-Darby canine kidney (MDCK) cell line has been found to be susceptible to epsilon-toxin (D. W. Payne, E. D. Williamson, H. Havard, N. Modi, and J. Brown, FEMS Microbiol. Lett. 116:161-168, 1994). Here we present evidence that epsilon-toxin cytotoxic activity is correlated with the formation of a large membrane complex (about 155 kDa) and efflux of intracellular K+ without entry of the toxin into the cytosol. Epsilon-toxin induced swelling, blebbing, and lysis of MDCK cells. Iodolabeled epsilon-toxin bound specifically to MDCK cell membranes at 4 and 37 labeled C and was associated with a large complex (about 155 kDa). The binding of epsilon-toxin to the cell surface was corroborated by immunofluorescence staining. The complex formed at 37 degrees C was more stable than that formed at 4 degrees C, since it was not dissociated by 5% sodium dodecyl sulfate and boiling.
ε毒素由B型和D型产气荚膜梭菌产生,可导致动物迅速致命的肠毒血症,其特征是由于血管通透性增加,多个器官出现水肿。已发现Madin-Darby犬肾(MDCK)细胞系对ε毒素敏感(D. W. Payne、E. D. Williamson、H. Havard、N. Modi和J. Brown,《FEMS微生物学快报》116:161 - 168,1994年)。在此,我们提供证据表明,ε毒素的细胞毒性活性与一种大的膜复合物(约155 kDa)的形成以及细胞内K⁺的外流相关,而毒素并未进入细胞质。ε毒素诱导MDCK细胞肿胀、起泡和裂解。碘标记的ε毒素在4℃和37℃时特异性结合到MDCK细胞膜上,并与一种大复合物(约155 kDa)相关。免疫荧光染色证实了ε毒素与细胞表面的结合。37℃形成的复合物比4℃形成的复合物更稳定,因为它不会被5%的十二烷基硫酸钠解离和煮沸。