NR AXRY
AU Mahal,S.P.; Demczyk,C.A.; Ercan,D.; Weissmann,C.
TI Discrimination of Prion Strains by the Cell Panel Assay
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Oral Abstracts FC2.1
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Vortrag
AB
Cell lines can be highly heterogeneous with respect to their susceptibility to different prion strains. For example, N2a-PK1 cells can propagate the strains RML and 22L but not Me7 and 87V. The sister clone N2a-R33 can propagate 22L but not RML. We set out to establish a cell-based assay to discriminate between prion strains in cell culture. Subclones of various cell lines were exposed to different prion strains and assayed by the Scrapie Cell Assay (SCA) (Klöhn et al. PNAS; 100: 11666-71; 2003). Clones showing high susceptibility to one or more of the strains were isolated and subjected to further rounds of subcloning and selection. Four cell lines showing different responses to the various strains were assembled into a panel. The panel was exposed to a dilution series of strains and the Response Index (RI) was determined for each strain. The RI on the different cell lines revealed a distinct strain signature allowing us to differentiate between prion strains 22L, RML, Me7 and the mouse-adapted BSE strain 301C. Using the panel assay, we found that in some cases propagation of prions in cell culture, for example 22L in R33, leads to a change of their characteristic signature.
The different characteristics after passage in cell culture may be a result of a change in the strain or due to a transient modification imparted by the cells, such as the glycosylation pattern of the PrP or some associated cell component. The modified prions have been inoculated into mice and will be assayed on the cell panel to see whether the modified characteristics are maintained or revert back to that of the original strain.
AD S.P. Mahal, C.A. Demczyk, D. Ercan, C. Weissmann, Scripps Florida, Infectology, USA
SP englisch
PO Schottland