title: A transfectant RK13 cell line permissive to classical caprine scrapie prion propagation.
abstract: To assess scrapie infectivity associated with caprine-origin tissues, bioassay can be performed using kids, lambs or transgenic mice expressing caprine or ovine prion (PRNP) alleles, but the incubation periods are fairly long. Although several classical ovine scrapie prion permissive cell lines with the ability to detect brain-derived scrapie prion have been available, no classical caprine scrapie permissive cell line is currently available. Therefore, the aims of this study were to generate a rabbit kidney epithelial cell line (RK13) stably expressing caprine wild-type PRNP (cpRK13) and then to assess permissiveness of cpRK13 cells to classical caprine scrapie prion propagation. The cpRK13 and plasmid control RK13 (pcRK13) cells were incubated with brain-derived classical caprine scrapie inocula prepared from goats or ovinized transgenic mice (Tg338, express ovine VRQ allele) infected with caprine scrapie. Significant PrP(Sc) accumulation, which is indicative of scrapie prion propagation, was detected by TSE ELISA and immunohistochemistry in cpRK13 cells inoculated with classical caprine scrapie inocula. Western blot analysis revealed the typical proteinase K-resistant 3 PrP(res) isoforms in the caprine scrapie prion inoculated cpRK13 cell lysate. Importantly, PrP(Sc) accumulation was not detected in similarly inoculated pcRK13 cells, whether by TSE ELISA, immunohistochemistry, or western blot. These findings suggest that caprine scrapie prions can be propagated in cpRK13 cells, thus this cell line may be a useful tool for the assessment of classical caprine prions in the brain tissues of goats.
authors.affiliation: a Department of Veterinary Microbiology and Pathology , College of Veterinary Medicine, Washington State University , Pullman , WA , USA ;
authors.forename: Rohana P
authors.initials: RP
authors.lastname: Dassanayake
authors.affiliation: b Animal Disease Research Unit, Agricultural Research Service , U.S. Department of Agriculture , Pullman , WA , USA.
authors.forename: Dongyue
authors.initials: D
authors.lastname: Zhuang
authors.affiliation: b Animal Disease Research Unit, Agricultural Research Service , U.S. Department of Agriculture , Pullman , WA , USA.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Truscott
authors.affiliation: a Department of Veterinary Microbiology and Pathology , College of Veterinary Medicine, Washington State University , Pullman , WA , USA ;
authors.forename: Sally A
authors.initials: SA
authors.lastname: Madsen-Bouterse
authors.affiliation: a Department of Veterinary Microbiology and Pathology , College of Veterinary Medicine, Washington State University , Pullman , WA , USA ;
authors.forename: Katherine I
authors.initials: KI
authors.lastname: O'Rourke
authors.affiliation: a Department of Veterinary Microbiology and Pathology , College of Veterinary Medicine, Washington State University , Pullman , WA , USA ;|b Animal Disease Research Unit, Agricultural Research Service , U.S. Department of Agriculture , Pullman , WA , USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Schneider
mesh_terms: D000818:Animals; D002460:Cell Line; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology*; D015870:Gene Expression; D015511:Goat Diseases / Q000235:genetics / Q000473:pathology*; D006041:Goats / Q000235:genetics*; D000072002:Prion Proteins / Q000032:analysis / Q000235:genetics*; D011817:Rabbits; D012608:Scrapie / Q000235:genetics / Q000473:pathology*; D014162:Transfection
chemical_list: D000072002:Prion Proteins
keywords: Caprine; Cell culture; Classical scrapie; Prions; Propagation; RK13 cells