Correlation between in vitro cytotoxicity and in vivo lethal activity in mice of epsilon toxin mutants from Clostridium perfringens

Epsilon toxin (Etx) from Clostridium perfringens is a pore-forming protein with a lethal effect on livestock, producing severe enterotoxemia characterized by general edema and neurological alterations. Site-specific mutations of the toxin are valuable tools to study the cellular and molecular mechan...

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Bibliographic Details
Published in:PloS one 2014-07, Vol.9 (7), p.e102417
Main Authors: Dorca-Arévalo, Jonatan, Pauillac, Serge, Díaz-Hidalgo, Laura, Martín-Satué, Mireia, Popoff, Michel R, Blasi, Juan
Format: Article
Language:English
Subjects:
Analysis
Animals
Apoptosis
Bacteria
Bacterial Toxins
Bacterial Toxins - genetics
Bacterial Toxins - metabolism
Bacterial Toxins - toxicity
Biocompatibility
Biological Transport
Biology and Life Sciences
Biomedical research
Blood-Brain Barrier
Blood-Brain Barrier - metabolism
Brain
Brain - drug effects
Brain - metabolism
Brain - pathology
Brain research
Cell membranes
Cellular Biology
Clostridium Infections
Clostridium Infections - microbiology
Clostridium Infections - mortality
Clostridium Infections - physiopathology
Clostridium perfringens
Clostridium perfringens - genetics
Clostridium perfringens - growth & development
Clostridium perfringens - pathogenicity
Correlation
Cysteine
Cytotoxicity
Cèl·lules epitelials
Dogs
Edema
Enterotoxemia
Enterotoxemia - microbiology
Enterotoxemia - mortality
Enterotoxemia - physiopathology
Epithelial Cells
Epithelial Cells - drug effects
Epithelial Cells - metabolism
Epithelial Cells - pathology
Experiments
Fluorescence
Fusion protein
Gene Expression
Green fluorescent protein
Green Fluorescent Proteins
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
In vivo methods and tests
Kidney Tubules, Distal
Kidney Tubules, Distal - drug effects
Kidney Tubules, Distal - metabolism
Kidney Tubules, Distal - pathology
Laboratories
Life Sciences
Livestock
Madin Darby Canine Kidney Cells
Male
Medicine and Health Sciences
Membranes cel·lulars
Mice
Mutants
Mutation
Neurosciences
Ovis aries
Parenchyma
Pathology
Pore formation
Primary Cell Culture
Proteins
Recombinant Fusion Proteins
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Recombinant Fusion Proteins - toxicity
Structure-Activity Relationship
Studies
Survival Analysis
Toxicity
Toxicology
Toxines
Toxins
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Summary:Epsilon toxin (Etx) from Clostridium perfringens is a pore-forming protein with a lethal effect on livestock, producing severe enterotoxemia characterized by general edema and neurological alterations. Site-specific mutations of the toxin are valuable tools to study the cellular and molecular mechanism of the toxin activity. In particular, mutants with paired cysteine substitutions that affect the membrane insertion domain behaved as dominant-negative inhibitors of toxin activity in MDCK cells. We produced similar mutants, together with a well-known non-toxic mutant (Etx-H106P), as green fluorescent protein (GFP) fusion proteins to perform in vivo studies in an acutely intoxicated mouse model. The mutant (GFP-Etx-I51C/A114C) had a lethal effect with generalized edema, and accumulated in the brain parenchyma due to its ability to cross the blood-brain barrier (BBB). In the renal system, this mutant had a cytotoxic effect on distal tubule epithelial cells. The other mutants studied (GFP-Etx-V56C/F118C and GFP-Etx-H106P) did not have a lethal effect or cross the BBB, and failed to induce a cytotoxic effect on renal epithelial cells. These data suggest a direct correlation between the lethal effect of the toxin, with its cytotoxic effect on the kidney distal tubule cells, and the ability to cross the BBB.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0102417