A transfectant RK13 cell line permissive to classical caprine scrapie prion propagation

Abstract

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.

Select Gene Expression Across Variable PrPSc Permissive Ovine Microglia

“…Potentially, prion protein could have a similar interaction with FN1. Additional studies of FN1 are warranted to determine if similar correlations with prion permissibility exist in other cell culture systems (i.e., Rov or cpRK13 [35]) and prion strains (i.e., natural scrapie isolates X124 or SSBP-1, Nor98, human prion isolates). Furthermore, future studies to silence FN1 or deletion of FN1 from culture media in order to create cell culture models for human prions may be warranted.…”

From Cell Culture to Organoids-Model Systems for Investigating Prion Strain Characteristics

“…Dassanayake et al, demonstrated that RK13 cells engineered to express caprine PrP with haplotype 2 (cpRK13 cells) could be infected with natural caprine scrapie of haplotype 1,1 or haplotype 1,2, but not to natural cases with haplotypes 3 or 4. However, Tg338 mice, which express caprine scrapie with haplotype 2, were able to propagate these natural haplotype 3 and 4 isolates [ 56 ]. Therefore, these mice might possess specific cofactors necessary for propagation that are lacking in RK13 cells.…”

“…Therefore, these mice might possess specific cofactors necessary for propagation that are lacking in RK13 cells. Subsequently, they found that after 2 serial passages of haplotype 3 or 4 scrapie isolates in Tg338 mice, cpRK13 cells were able to propagate these mouse-adapted isolates [ 56 ].…”

Cellular models for discovering prion disease therapeutics: Progress and challenges

“…RK13 cells were one of the first cell lines to replicate non-murineadapted prion strains, and the first to replicate natural ovine scrapie (Vilette et al, 2001). RK13 cells have proven to be an impressively versatile cell line given that they have been shown thus far to replicate ovine scrapie, caprine scrapie, vole-adapted BSE, CWD, and mouse prions upon expression of the appropriate PrP C molecule (Bian et al, 2010;Courageot et al, 2008;Dassanayake et al, 2016;Grassmann, Wolf, Hofmann, Graham, & Vorberg, 2013;Vilette et al, 2001). This versatility not only allows for the study of each prion disease individually but also offers researchers the ability to compare and study vari- Studying scrapie-infected MovS, in particular, may allow for greater insight into the processes by which prions invade the peripheral nervous system (Bedecs, 2008 (Archer et al, 2004;Neale et al, 2010;Sabuncu et al, 2003;Vilette et al, 2001).…”

“…One cell line, RK13 cells transfected with caprine PrP C (cpRK13), has been reported to successfully and stably replicate goat scrapie prions (Dassanayake et al, 2016). Although a caprine scrapie-infected cell line derived from goat tissue would better mirror real-life conditions, previous attempts to establish such a model have failed (Oelschlegel et al, 2015).…”