NR AWGT
AU Heinig,L.; Koelbel,K.; Mueller,D.A.; Hunsmann,G.; Holznagel,E.; Stuke,A.W.
TI Infection of PrP knock-out cells conditionally expressing mammalian prion proteins
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-18
PT Konferenz-Poster
AB The prion protein (PrP) can be the cause of TSE disease, including mad cow disease (BSE), chronic wasting disease (CDW), and Creutzfeldt-Jakob disease (CJD). These diseases are resulting from conversion of a cellular non-pathogenic to an infective pathogenic isoform. For understanding prion diseases the function of the normal cellular isoform is a fundamental must have. We created a prion expression system in murine PrP knock-out cells. In these cells the stable transfected prnp gene was changed between different mammals, including human, chimp, crab eating monkey, cow and moose. In the different cell lines PrP expression is regulated by a tetracyline responding element (TRE). The regulation was achieved by using different tetracycline or doxycxline concentrations. The expression was analysed by western blot (Wb), FACS, immunfluorescence (IF) and circular dichroism (CD) spectroscopy analyses. After characterization of PrP expression, the cells lines were infected with different concentrations of a BSE stem brain homogenate. PrP propagation in the cells was observed after 48 h by using Wb, IF, and FACS analyses. The CD spectroscopy analysis of infected cells is still in process. In addition we will do kinetic studies including proof of chronic infection. From our observations we conclude that PrP knock-out cells stably transfected with human, primate, and ruminant prnp can be infected with BSE. Currently, the molecular mechanism of prion protein propagation is examined more accurately by using 2-D gel electrophoresis (2-DE) analysis of infected and non-infected cell proteome. This system can be used not only for infection studies instead of animal models. It can also be used for characterization of the molecular mechanisms of prion infectivity, e.g. the signal cascade of other proteins they are directly or indirectly involved in the prion protein expression and also PrP conversion.
AD L. Heinig, D.A. Mueller, G. Hunsmann, A.W. Stuke: Dept. Virology and Immunology, German Primate Centre, 37073 Göttingen, Germany; K. Koelbel: Institute for Biochemistry, Martin-Luther-University Halle-Wittenberg, 06120 Halle, Germany; E. Holznagel: Primate Center, Paul-Ehrlich-Institut, 63225 Langen, Germany
SP englisch
PO Italien