title: Correlation between in vitro cytotoxicity and in vivo lethal activity in mice of epsilon toxin mutants from Clostridium perfringens.
abstract: 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.
authors.affiliation: Laboratory of Cellular and Molecular Neuroscience, Department of Pathology and Experimental Therapeutics, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain; IDIBELL-Bellvitge Biomedical Research Institute, L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: Jonatan
authors.initials: J
authors.lastname: Dorca-Arévalo
authors.affiliation: Institut Pasteur, Unité des Bactéries anaérobies et Toxines, Paris, France.
authors.forename: Serge
authors.initials: S
authors.lastname: Pauillac
authors.affiliation: Laboratory of Cellular and Molecular Neuroscience, Department of Pathology and Experimental Therapeutics, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Díaz-Hidalgo
authors.affiliation: Laboratory of Cellular and Molecular Neuroscience, Department of Pathology and Experimental Therapeutics, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain; IDIBELL-Bellvitge Biomedical Research Institute, L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Martín-Satué
authors.affiliation: Institut Pasteur, Unité des Bactéries anaérobies et Toxines, Paris, France.
authors.forename: Michel R
authors.initials: MR
authors.lastname: Popoff
authors.affiliation: Laboratory of Cellular and Molecular Neuroscience, Department of Pathology and Experimental Therapeutics, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain; IDIBELL-Bellvitge Biomedical Research Institute, L'Hospitalet de Llobregat, Barcelona, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Blasi
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000378:metabolism; D001921:Brain / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D003015:Clostridium Infections / Q000382:microbiology / Q000401:mortality* / Q000503:physiopathology; D003016:Clostridium perfringens / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D004285:Dogs; D004767:Enterotoxemia / Q000382:microbiology / Q000401:mortality* / Q000503:physiopathology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007686:Kidney Tubules, Distal / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D061985:Madin Darby Canine Kidney Cells; D008297:Male; D051379:Mice; D009154:Mutation; D061251:Primary Cell Culture; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D013329:Structure-Activity Relationship; D016019:Survival Analysis
chemical_list: D001427:Bacterial Toxins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins