NR AWKO
AU Moonen,P.; Ruiter,S.; Bossers,A.
TI Eukaryotic cells expressing heterologous PrPc as a putative tool in the diagnosis of prion diseases
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions CE-32
PT Konferenz-Poster
AB Currently, several different strains of pathologically folded prion protein (PrPsc) are recognised in sheep and cattle. Different strains are ultimately defined by bioassay, but can also be determined by more rapid methods like migration behaviour of the pK resistant fragment of PrPsc or by interpretation of the PrPsc distribution in the brain of infected animals. The research presented here focuses on enhancing the ability to discriminate between strains using eukaryotic cells expressing different variants of the cellular form of the prion protein (bovine and several allelic variants ovine). Several isolates of scrapie and BSE have been used for in vitro infection of the PrPc expressing cells available. Cells were serially passaged weekly for several weeks and accumulation of PrPsc has been determined using Western blot. Distribution of PrPsc was studied using immunoperoxidase monolayer assay (IPMA) and fluorescence microscopy. Persistent in vitro infection, as demonstrated in ROV and MOV cells (Vilette et al, 2001, Archer et al, 2004), could not be detected in our own cells. Transient infection was seen in Western blot, fluorescence microscopy, and IPMA, but did not allow the discrimination between different strains. Whether confocal laser scanning microscopy can enhance the resolution is currently being investigated.
AD Central Institute for Animal Disease Control (CIDC-Lelystad), The Netherlands. E-mail: peter.moonen@wur.nl
SP englisch
PO Italien