NR AWRB
AU Zanusso,G.; Mazza,M.; Farinazzo,A.; Acutis,P.L.; Casalone,C.; Martucci,F.; Fasoli,E.; Brocchi,E.; Capucci,L.; Barbieri,I.; Nonno,R.; Agrimi,U.; Monaco,S.; Caramelli,M.
TI Characterization of PrPres by two dimensional analysis in atypical scrapie cases in Italy
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-38
PT Konferenz-Poster
AB
In the last few years, active survelliance system of TSEs in cattle and in small ruminants induced the detection of an increasing number of scrapie and BSE cases characterized by a PrPsc differing from that of typical cases. Most of these "discordant" cases were positive to ELISA (Biorad) screening test and negative to immunoblot because of the low resistance of PrPsc to proteases treatment. In this study, we obtained brain samples from Italian scrapie cases of "Nor-98 like" in sheep and we carried out a biochemical characterization of PK-resistant PrP. One D-PAGE showed two different PrPsc banding patterns based on the anti-PrP antibody probed. In particular, by using mAb 4G11 directed to a C-terminal epitope (residues: 199-218) PrPsc was represented by three bands which migrated between 16-25kDa that were reduced to single band of 16kDa after deglycosylation. Conversely, P4 (residues: 89-104 ) detected a single band of 12kDa, that did not change in migration after glycans removal, likely representing N- and C-terminally truncated fragment. More interestingly, 2D-PAGE analysis showed that C-terminal and the internal fragments, recognized by mAbs 4G11 and P4 respectively, were clearly distinguishable because of to the opposite charges of this peptides. The 16 kDa C-terminal fragment had a pI ranging from 3.0 to 6.0 whereas the internal peptide of 12kDa migrated around a pH of 9.0. In addition, we compared 2D maps of the present cases to those obtained in previously described scrapie cases (Zanusso et al., J Gen Virol 2003). In conclusion, biochemical analysis of PrPsc in atypical scrapie cases showed that: (i) Italian cases show an electrophoretic PrP profile similar to those reported in other countries; (ii) the 16 and 12kDa fragments derive from two differently truncated fragments; (iii) PrPsc similarities, obtained by 2D-PAGE, between the present cases and previously reported scrapie cases suggest that the PrPsc pattern found in atypical cases might be frequently found in sheep population.
This work was supported by Neuroprion (FOOD-CT-2004-506579)
AD G. Zanusso, A. Farinazzo, E. Fasoli, S. Monaco: Department of Neurological and Visual Sciences, University of Verona, Italy; M. Mazza, P.L. Acutis, C. Casalone, F. Martucci, M. Caramelli: IZSPLVA, Turin, Italy; E. Brocchi, L. Capucci, I. Barbieri: IZSLER, Brescia, Italy; R. Nonno, U. Agrimi: ISS, Rome; Italy. E-mail: gianluigi.zanusso@univr.it
SP englisch
PO Italien