NR AWMC
AU Polak,M.P.; Zmudzinski,J.F.; Larska,M.; van Zijderveld,F.G.; Erkens,J.H.F.; van Keulen,L.J.M.; Stack,M.J.; Langeveld,J.P.M.
TI Unusual migration pattern of PrPres from bovine brainstem tissue
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 PR-28
PT Konferenz-Poster
AB Confirmation of BSE may be difficult in rare cases especially when quality of brainstem is poor as in dead-on-farm animals. Experience from Western blotting (WB) has revealed that a low speed clarifying centrifugation can help to confirm a negative diagnosis by the removal of equivocal PrP-signals. However, we present here three cases - 2 autolysed cases from Poland and a case from a slaughterhouse in The Netherlands - which after centrifugation persistenly exhibit PK-resistant PrP-immuno reactive bands with an unusual migration. Confirmation was performed by OIE-SAF immunoblotting in the autolysed samples and by IHC in the case from slaughterhouse. The objective of this study was to further examine these cases for molecular properties of the PrP-bands. Studies carried out were: graded digestion with PK, deglycosylation with PNGaseF, precipitations, analysis by WB with a range of monoclonal antibodies. In comparison with BASE-like (or L-type) isolates, the present 3 examples exhibit PrPres bands with lower MWr. Antibodies 12B2 and 9A2, specific for the respective PK-resistant N-terminal regions aa residues 101-105 and 108-110 of bovine PrP, were not reactive, and removal of carbohydrates by PNGaseF indicated that the molecular entity of the PrPres bands represented a glycosylated PrP-moiety encompassing an N-terminus located somewhere between aa residues 110 and 120 and the C-terminus of mature PrP. These observations call for a cautious approach especially when testing samples in a very advanced stage of autolysis. However, the finding of a similar case from a slaughterhouse indicates that all 3 represent natural in vivo phenomena rather than a consequence of autolysis.
AD M.P. Polak, J.F. Zmudzinki, M. Larska: National Veterinary Research Institute, Department of Virology, Pulawy, Poland; F.G. van Zijderveld, J.H.F. Erkens, L.J.M. van Keulen, J.P.M. Langeveld: CIDC-Lelystad, Department of Bacteriology and TSEs, Lelystad, The Netherlands; M.J. Stack: VLA, TSE Molecular Biology Department, Woodham Lane, Addlestone, Surrey UK. E-mail: ppolak@piwet.pulawy.pl
SP englisch
PO Italien