NR AOQO
AU Boesen,M.; Dietrich,R.; Groschup,M.H.; Pichner,R.; Gareis,M.; Müller,S.; Scherer,S.; Märtlbauer,E.
TI Distribution of cellular Prion Protein in selected tissues of ruminants
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-48
PT Konferenz-Poster
AB
As cellular Prion Protein (PrPc) acts as source for PrPsc replication, knowledge about the occurrence of PrPc in different tissues is of fundamental importance to enable a reasonable assessment of their potential infectivity. Astonishingly, only sporadic reports exist about the physiological tissue distribution of the PrPc in sheep and particularly in cattle.
With respect to the analysis of different tissues for PrPc, a rapid and sensitive immunochemical method using monoclonal antibodies (Mab) was established. Several Mab combinations were studied and a non-competitive sandwich system using Mab P4 for coating microtiter plates and a horseradish peroxidase labelled Mab, specific for the C-terminus, for detection turned out to be the most sensitive assay. For extraction of the different tissues, derived from lambs, calves and adult cattle, a simple and rapid method was established. Samples were homogenised in PBS containing 3% glucose by using a Ribolyser. The resulting homogenates were further diluted in PBS containing 1% BSA and then directly analysed by the sandwich EIA.
In sheep high levels were generally found in the brainstem, but considerable amounts of PrPc could also be detected in lungs, tonsils and third eye lid, whereas for muscles (skeleton as well as heart), most of the lymph nodes, thymus, liver, udder, ovaries and spleen only weak positive results were obtained. Varying reactivity could be found in skeleton muscle, heart and liver of calves and adult cattle.
AD M. Boesen, R. Dietrich, E. Märtlbauer, Ludwig-Maximilians-Universität München, Germany; M. Groschup, Institut für neue und neuartige Tierseuchenerreger, BFAV, Isle of Riems, Germany; R. Pichner, M. Gareis, Institut für Mikrobiologie und Toxikologie, BAFF, Kulmbach, Germany; S. Müller, S. Scherer, Institut für Mikrobiologie, ZIEL Weihenstephan, Freising, Germany
SP englisch
PO Deutschland