NR AXWK
AU Siso,S.; Jeffrey,M.; Martin,S.; McGovern,G.; Steele,P.; Finlayson,J.; Chianini,F.; Gonzalez,L.
TI Detection of PrPd in Kidneys from Scrapie-Affected Sheep
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.132
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Our preliminary findings on deposition of disease-associated PrP (PrPd) in kidneys from scrapie-affected sheep1 indicated for the first time that 1) PrPd accumulated in a significant proportion of kidneys from sheep naturally and experimentally exposed to scrapie, (2) PrPd deposition was consistently restricted to the renal papillae in close contact with the renal pelvis, 3) most immunolabelling was intracellular, and 4) accumulation of PrPd in kidneys occurred regardless of any concurrent renal inflammatory process.
The main purposes of this presentation are to 1) assess the specificity and sensitivity of different PrP antibodies, 2) describe the location and cell type-association of PrPd in the interstitium of renal papillae, and 3) to discuss the factors that might determine the renal deposition of PrPd and the risk of resulting environmental contamination.
We have examined kidneys collected at post-mortem from 24 naturally-infected ARQ/ARQ Suffolk sheep, and from 34 Suffolk or Cheviot sheep experimentallychallenged by the oral or subcutaneous route. The brain inocula used for these last was either from a Suffolk flock (n=22) or from a Cheviot flock (n=12), both with natural scrapie.
Detection of PrPd in the kidney by immunohistochemistry (IHC) using the antibody R145 was achieved in 25% (6/24) of the natural scrapie cases, in 68% (15/22) of the sheep dosed with the Suffolk source, and in 8% (1/12) of those challenged with the Cheviot source. Specificity and sensitivity of IHC results were checked by examining 11 kidneys from scrapie-free sheep, and by performing IHC with other PrP antibodies 8G8, 2A11, 2G11, 6H4, BG4, 12B2 and F99, respectively. In addition, kidney samples were examined by Western blotting with the 6H4 or P4 antibody, and the characteristic three-band pattern of PrPres, was confirmed in the majority of the IHC positive kidneys. The precise location of PrPd within the renal papilla was determined by double IHC for different cell markers and by immunoelectron microscopy; these studies confirmed that PrPd accumulated in the interstitium of the renal papillae, and was either intra-lysosomal or extracellular in close contact with basal membranes.
1 Sisó S, González L, Jeffrey M, Martin S, Chianini F, Steele P (2006) Prion protein in kidneys of scrapie-infected sheep. Vet Rec (letter), 159:327-8.
AD S. Sisó, M. Jeffrey, S. Martin, G. McGovern, L. González, VLA Lasswade, Pathology, UK; P. Steele, J. Finlayson, F. Chianini, Pentlands Science Park, Moredun Research Institute, UK
SP englisch
PO Schottland