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The effects of Clostridium perfringens enterotoxin on morphology, viability, and macromolecular synthesis in Vero
Abstract:
Vero (African green monkey kidney) cells grown in tissue culture monolayer were sensitive to Clostridium perfringens enterotoxin. Within 30 minutes of exposure to the enterotoxin gross morphological damage was observed and within 40 minutes approximately 75% of the cells had detached. Nearly half of the cells were nonviable following 35 to 40 minutes incubation with the enterotoxin. Doses as low as 0.1 ng caused small but detectable inhibition of plating efficiency of the cells while more than 100 ng caused the inhibition to approach 100%. Total inhibition of DNA, RNA, and protein synthesis occurred within 30 minutes exposure to enterotoxin. Heat inactivated enterotoxin had no apparent effects upon cellular morphology, detachment, viability, plating efficiency, or incorporation. We propose that the enterotoxin induces structural damage to the cytoplasmic membrane which results in loss of electrolytes and other essential substances from the cells. The outcome of this process is shut down of macromolecular synthesis, gross morphological damage, and eventual death of the cell.
Insights
Clostridium perfringens enterotoxin rapidly damages Vero cells, causing detachment, death, and halting macromolecular synthesis. Heat-inactivated toxin showed no adverse effects, supporting its role in cell membrane damage.
Area of Science:
- Microbiology
- Cell Biology
- Toxicology
Background:
- Clostridium perfringens is a common cause of foodborne illness.
- Enterotoxins produced by C. perfringens are responsible for gastrointestinal symptoms.
- The precise mechanism of enterotoxin-induced cell damage requires further elucidation.
Purpose of the Study:
- To investigate the cytotoxic effects of Clostridium perfringens enterotoxin on Vero cells.
- To determine the time course and dose-dependency of enterotoxin-induced cellular damage.
- To elucidate the molecular mechanisms underlying enterotoxin-mediated cell death.
Main Methods:
- Vero cells were cultured in monolayers and exposed to varying concentrations of C. perfringens enterotoxin.
- Cellular morphology, detachment, viability, and plating efficiency were assessed.
- DNA, RNA, and protein synthesis were measured using incorporation assays.
- Heat-inactivated enterotoxin was used as a control.
Main Results:
- Enterotoxin exposure led to rapid morphological damage and cell detachment within 30-40 minutes.
- Cell viability decreased significantly, with nearly 50% nonviable cells after 40 minutes.
- Low enterotoxin doses (0.1 ng) inhibited plating efficiency, while high doses (>100 ng) caused near-complete inhibition.
- Total inhibition of DNA, RNA, and protein synthesis occurred within 30 minutes.
- Heat-inactivated enterotoxin had no observable effects.
Conclusions:
- Clostridium perfringens enterotoxin induces rapid and severe damage to Vero cells.
- The enterotoxin likely targets the cytoplasmic membrane, leading to loss of essential cellular components.
- This membrane damage results in the cessation of macromolecular synthesis and eventual cell death.