title: Killing of Trypanozoon Parasites by the Equine Cathelicidin eCATH1.
abstract: Trypanozoon parasites infect both humans, causing sleeping sickness, and animals, causing nagana, surra, and dourine. Control of nagana and surra depends to a great extent on chemotherapy. However, drug resistance to several of the front-line drugs is rising. Furthermore, there is no official treatment for dourine. Therefore, there is an urgent need to develop antiparasitic agents with novel modes of action. Host defense peptides have recently gained attention as promising candidates. We have previously reported that one such peptide, the equine antimicrobial peptide eCATH1, is highly active against equine Gram-positive and Gram-negative bacteria, without cytotoxicity against mammalian cells at bacteriolytic concentrations. In the present study, we show that eCATH1 exhibits an in vitro 50% inhibitory concentration (IC50) of 9.5 μM against Trypanosoma brucei brucei, Trypanosoma evansi, and Trypanosoma equiperdum Its trypanocidal mechanism involves plasma membrane permeabilization and mitochondrial alteration based on the following data: (i) eCATH1 induces the rapid influx of the vital dye SYTOX Green; (ii) it rapidly disrupts mitochondrial membrane potential, as revealed by immunofluorescence microscopy using the fluorescent dye rhodamine 123; (iii) it severely damages the membrane and intracellular structures of the parasites as early as 15 min after exposure at 9.5 μM and 5 min after exposure at higher concentrations (19 μM), as evidenced by scanning and transmission electron microscopy. We also demonstrate that administration of eCATH1 at a dose of 10 mg/kg to T. equiperdum-infected mice delays mortality. Taken together, our findings suggest that eCATH1 is an interesting template for the development of novel therapeutic agents in the treatment of trypanosome infections.
authors.affiliation: ANSES, Dozule Laboratory for Equine Diseases, Bacteriology and Parasitology Unit, Goustranville, France Université de Caen Basse-Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France.
authors.forename: S
authors.initials: S
authors.lastname: Cauchard
authors.affiliation: Unit of Parasite Diagnostics, Department of Biomedical Sciences, Institute for Tropical Medicine, Antwerp, Belgium.
authors.forename: N
authors.initials: N
authors.lastname: Van Reet
authors.affiliation: Unit of Parasite Diagnostics, Department of Biomedical Sciences, Institute for Tropical Medicine, Antwerp, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: Büscher
authors.affiliation: Centre de Microscopie Appliquée à la Biologie, SF4206 ICORE, Université de Caen Basse-Normandie, France.
authors.forename: D
authors.initials: D
authors.lastname: Goux
authors.affiliation: Institute of Biochemistry, Christian-Albrechts University of Kiel, Kiel, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Grötzinger
authors.affiliation: Zoological Institute, Comparative Immunobiology, Christian-Albrechts University of Kiel, Kiel, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Leippe
authors.affiliation: Université de Caen Basse-Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France CHU de Caen, Service de Microbiologie, Caen, France CNR de la Résistance aux Antibiotiques, Laboratoire Associé Entérocoques, Caen, France.
authors.forename: V
authors.initials: V
authors.lastname: Cattoir
authors.affiliation: ANSES, Dozule Laboratory for Equine Diseases, Bacteriology and Parasitology Unit, Goustranville, France.
authors.forename: C
authors.initials: C
authors.lastname: Laugier
authors.affiliation: ANSES, Dozule Laboratory for Equine Diseases, Bacteriology and Parasitology Unit, Goustranville, France julien.cauchard@anses.fr.
authors.forename: J
authors.initials: J
authors.lastname: Cauchard
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D020128:Inhibitory Concentration 50; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014345:Trypanosoma / Q000187:drug effects*; D014346:Trypanosoma brucei brucei / Q000187:drug effects*
chemical_list: D023181:Antimicrobial Cationic Peptides; D014344:Trypanocidal Agents
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