a simple, versatile and sensitive cell-based assay for prions from various species一个简单的,通用的和敏感的细胞试验对朊病毒从不同的物种.pdfVIP

A Simple, Versatile and Sensitive Cell-Based Assay for Prions from Various Species Zaira E. Arellano-Anaya, Jimmy Savistchenko, Jacinthe Mathey, Alvina Huor, Caroline Lacroux, Olivier ´ Andreoletti, Didier Vilette* ´ ´ ˆ ` UMR INRA (Institut National de la Recherche Agronomique) ENVT (Ecole Nationale Veterinaire de Toulouse) 1225, Interactions Hote Agent Pathogene, Toulouse, France 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. Citation: Arellano-Anaya ZE, Savistchenko J, Mathey J, Huor A, Lacr