title: A simple, versatile and sensitive cell-based assay for prions from various species.
abstract: Detection and quantification of prion infectivity is a crucial step for various fundamental and applied aspects of prion research. Identification of cell lines highly sensitive to prion infection led to the development of cell-based titration procedures aiming at replacing animal bioassays, usually performed in mice or hamsters. However, most of these cell lines are only permissive to mouse-adapted prions strains and do not allow titration of prions from other species. In this study, we show that epithelial RK13, a cell line permissive to mouse and bank vole prion strains and to natural prion agents from sheep and cervids, enables a robust and sensitive detection of mouse and ovine-derived prions. Importantly, the cell culture work is strongly reduced as the RK13 cell assay procedure designed here does not require subcultivation of the inoculated cultures. We also show that prions effectively bind to culture plastic vessel and are quantitatively detected by the cell assay. The possibility to easily quantify a wider range of prions, including rodent experimental strains but also natural agents from sheep and cervids, should prompt the spread of cell assays for routine prion titration and lead to valuable information in fundamental and applied studies.
authors.affiliation: UMR Institut National de la Recherche Agronomique Ecole Nationale Vétérinaire de Toulouse 1225, Interactions Hôte Agent Pathogène, Toulouse, France.
authors.forename: Zaira E
authors.initials: ZE
authors.lastname: Arellano-Anaya
authors.forename: Jimmy
authors.initials: J
authors.lastname: Savistchenko
authors.forename: Jacinthe
authors.initials: J
authors.lastname: Mathey
authors.forename: Alvina
authors.initials: A
authors.lastname: Huor
authors.forename: Caroline
authors.initials: C
authors.lastname: Lacroux
authors.forename: Olivier
authors.initials: O
authors.lastname: Andréoletti
authors.forename: Didier
authors.initials: D
authors.lastname: Vilette
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D015151:Immunoblotting; D051379:Mice; D008822:Mice, Transgenic; D011328:Prions / Q000032:analysis*; D012756:Sheep
chemical_list: D011328:Prions