NR AQRU
AU Piccardo,P.; Tagliavini,F.; Dlouhy,S.R.; Young,K.; Seiler,C.; Rossi,G.; Hill,A.F.; Bugiani,O.; Collinge,J.; Ghetti,B.
TI An antiserum to residues 95-108 of human PrP detects PrPres in a variety of human and animal prion diseases
QU Journal of Neuropathology and Experimental Neurology 1997; 56(5): 589 Nr. 73
PT Meeting Abstract
VT The molecular hallmark of the prion diseases is the presence of protease resistant prion protein (PrPres) in the central nervous system. It has been proposed that PrPres derives from normal PrP by post-translational modifications that may result in PrP isoforms with different physicochemical properties. We synthesized peptides corresponding to the human PrP sequences 23-40, 58-71, 90-102, 95-108, 151-165, 220-231, and raised polyclonal antibodies, using the multiple antigenic peptide system. Our aim is to develop a panel of antibodies that can recognize isoforms and related breakdown products. Upon characterization, we observed that most antisera show an endpoint titer of approximately 1:100,000. Immunohistochemically, the antibodies recognized PrP deposits with or without amyloid tinctorial properties. The region comprising residues 98-108 of human PrP is highly conserved among species including cow, goat, sheep, kudu, chinese hamster and mouse. We have carried out western immunoblot studies, using antiserum to residues 95-108 (PrP 95-108) at a 1:15,000/1:20,000 dilution. PrP 95-108 recognizes numerous isoforms of PrPres in (i) sporadic Creutzfeldt-Jakob disease (CJD), (ii) new variant CJD, (iii) bovine spongiform encephalopathy (BSE), (iv) sheep with natural scrapie, (v) hamster with experimental scrapie. In contrast, monoclonal antibody 3F4 which recognizes human PrP residues 109-112 does not detect PrP in brain extracts of BSE and natural scrapie. These findings indicate that distinct PrPres isoforms are detected in different diseases and in different animal species with a single antibody to PrP. Supported by N529822 'and the Italian Ministry of Health.
IN Während polyklonale Antikörper gegen von den hochkonservierten Aminosäuren 95-108 des Prionproteins abgeleitete Peptide proteaseresistentes Prionprotein in CJD- und vCJD-Patienten, Rind, Schaf und Hamster erkannten, markierten Antikörper gegen PrP-109-112 weder in Rind, noch in Schaf die proteaseresistenten Prionproteine.
ZR 0
AD P.Piccardo*1, F.Tagliavini*2, R.Dlouhy1, K.Young1, C.Seiler1, G.Rossi2, A.F.Hill3, M.Bugiani2, O.Bugiani*2, J.Collinge3, B.Ghetti*1, 1 Indiana University, Indianapolis, IN, USA; 2 Istituto Neurologico Carlo Besta, Milano, Italy; 3 Imperial College School of Medicine at St. Mary's, London, UK
SP englisch
OR Prion-Krankheiten 6