Effect of Epsilon Toxin–GFP on MDCK Cells and Renal Tubules In Vivo

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  • Alex Soler-Jover
    Departament de Biologia Cel·lular i Anatomia Patològica, Campus de Bellvitge, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain
  • Juan Blasi
    Departament de Biologia Cel·lular i Anatomia Patològica, Campus de Bellvitge, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain
  • Inma Gómez de Aranda
    Departament de Biologia Cel·lular i Anatomia Patològica, Campus de Bellvitge, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain
  • Piedad Navarro
    Departament de Biologia Cel·lular i Anatomia Patològica, Campus de Bellvitge, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain
  • Maryse Gibert
    CNR Anaérobies, Institut Pasteur, Paris, France
  • Michel R. Popoff
    CNR Anaérobies, Institut Pasteur, Paris, France
  • Mireia Martín-Satué
    Departament de Biologia Cel·lular i Anatomia Patològica, Campus de Bellvitge, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain

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<jats:p> Epsilon toxin (∊-toxin), produced by Clostridium perfringens types B and D, causes fatal enterotoxemia, also known as pulpy kidney disease, in livestock. Recombinant ∊-toxin–green fluorescence protein (∊-toxin–GFP) and ∊-prototoxin–GFP were successfully expressed in Escherichia coli. MTT assays on MDCK cells confirmed that recombinant ∊-toxin–GFP retained the cytotoxicity of the native toxin. Direct fluorescence analysis of MDCK cells revealed a homogeneous peripheral pattern that was temperature sensitive and susceptible to detergent. ∊-Toxin–GFP and ∊-prototoxin-GFP bound to endothelia in various organs of injected mice, especially the brain. However, fluorescence mainly accumulated in kidneys. Mice injected with ∊-toxin–GFP showed severe kidney alterations, including hemorrhagic medullae and selective degeneration of distal tubules. Moreover, experiments on kidney cryoslices demonstrated specific binding to distal tubule cells of a range of species. We demonstrate with new recombinant fluorescence tools that ∊-toxin binds in vivo to endothelial cells and renal tubules, where it has a strong cytotoxic effect. Our binding experiments indicate that an ∊-toxin receptor is expressed on renal distal tubules of mammalian species, including human. (J Histochem Cytochem 52:931–942, 2004) </jats:p>

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