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Assembly of Clostridium perfringens epsilon-toxin on MDCK cell membrane
M Nagahama1, S Ochi, J Sakurai
1Department of Microbiology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Japan.
Abstract:
Clostridium perfringens epsilon-toxin bound to the Madin Darby canine kidney (MDCK) cells and aggregated. The complex of the toxin was formed in a dose- and a time-dependent manner. The formation of the complex increased with a decrease in viable counts of MDCK cells and with increasing K+ release from the cells. The inactivated toxin heated at 100 degrees C did not aggregate under the condition. In addition, the prototoxin dose-dependently bound to the cells, but did not form the complex. Incubation of the toxin with MDCK cell membranes also showed the formation of the complex, but that with membrane preparations prepared from Vero cells or sheep erythrocytes, which are insensitive for the toxin, showed no formation of the complex. Incubation of the toxin with mouse brain homogenates resulted in formation of the complex, but that with brain homogenates heated at 80 degrees C or mouse liver homogenates showed no formation of the complex. These observations show that the complex formation of epsilon-toxin is essential for toxicity of the toxin.
Insights
Clostridium perfringens epsilon-toxin aggregation with susceptible cells, like Madin Darby canine kidney (MDCK) cells, is crucial for its toxicity. This complex formation correlates with cell damage and ion release.
Area of Science:
- Microbiology
- Toxicology
- Cell Biology
Background:
- Clostridium perfringens epsilon-toxin is a potent virulence factor.
- Understanding the mechanism of epsilon-toxin's action is vital for developing effective countermeasures.
Purpose of the Study:
- To investigate the complex formation of Clostridium perfringens epsilon-toxin with target cells.
- To determine the relationship between complex formation and toxin-induced cell damage.
Main Methods:
- Incubation of epsilon-toxin with Madin Darby canine kidney (MDCK) cells and cell membranes.
- Assessing toxin-cell complex formation via dose- and time-dependency.
- Monitoring cell viability and potassium (K+) release.
- Using heat-inactivated toxin and non-sensitive cell membranes/homogenates as controls.
Main Results:
- Epsilon-toxin formed a dose- and time-dependent complex with MDCK cells, correlating with decreased cell viability and increased K+ release.
- Heat-inactivated toxin and prototoxin did not form complexes.
- Complex formation occurred with MDCK cell membranes and mouse brain homogenates but not with insensitive cell membranes or liver homogenates.
Conclusions:
- Complex formation between epsilon-toxin and susceptible cell membranes is essential for its toxicity.
- The findings highlight a critical step in the epsilon-toxin's pathogenic mechanism.