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Effect of Clostridium perfringens epsilon toxin on MDCK cells
E Borrmann1, H Günther, H Köhler
1Federal Institute for Health Protection of Consumers and Veterinary Medicine, Division 4, Jena, Germany. e.borrmann@bgvv.de
Abstract:
Epsilon toxin is one of the major lethal toxins produced by Clostridium perfringens type D and B. It is responsible for a rapidly fatal disease in sheep and other farm animals. Many facts have been published about the physical properties and the biological activities of the toxin, but the molecular mechanism of the action inside the cells remains unclear. We have found that the C. perfringens epsilon toxin caused a significant decrease of the cell numbers and a significant enlargement of the mean cell volume of MDCK cells. The flow cytometric analysis of DNA content revealed the elongation of the S phase and to a smaller extent of the G2+M phase of toxin-treated MDCK cells in comparison to untreated MDCK cells. The results of ultrastructural studies showed that the mitosis is disturbed and blocked at a very early stage, and confirmed the toxin influence on the cell cycle of MDCK cells.
Insights
Clostridium perfringens epsilon toxin significantly reduces cell numbers and enlarges cell volume in MDCK cells. This toxin disrupts cell cycle progression, blocking mitosis and impacting cell division.
Area of Science:
- Microbiology
- Cell Biology
- Toxicology
Background:
- Epsilon toxin from Clostridium perfringens is a lethal agent in livestock.
- Its physical and biological properties are known, but the intracellular molecular mechanism remains elusive.
Purpose of the Study:
- To investigate the molecular mechanism of Clostridium perfringens epsilon toxin's action on cell cycle progression.
- To analyze the effects of epsilon toxin on Madin-Darby Canine Kidney (MDCK) cell proliferation and cell cycle phases.
Main Methods:
- Cell counting and mean cell volume analysis of MDCK cells.
- Flow cytometric analysis of DNA content to assess cell cycle distribution.
- Ultrastructural studies to examine cellular morphology and mitosis.
Main Results:
- Epsilon toxin treatment led to a significant decrease in MDCK cell numbers and an increase in mean cell volume.
- Flow cytometry revealed an elongated S phase and G2+M phase in toxin-treated cells.
- Ultrastructural analysis confirmed disturbed mitosis, blocked at an early stage, indicating cell cycle arrest.
Conclusions:
- Clostridium perfringens epsilon toxin significantly impacts MDCK cell proliferation and viability.
- The toxin interferes with cell cycle progression, specifically affecting DNA replication and mitosis.
- These findings elucidate a key aspect of epsilon toxin's molecular mechanism of action at the cellular level.
