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From: TSS ()
Subject: Molecular profiling of ovine prion diseases using thermolysin-resistant PrPSc and endogenous C2 PrP fragments
Date: July 26, 2007 at 6:59 am PST

J. Virol. doi:10.1128/JVI.00640-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.


Molecular profiling of ovine prion diseases using thermolysin-resistant PrPSc and endogenous C2 PrP fragments

Jonathan P. Owen, Helen C. Rees, Ben C. Maddison, Linda A. Terry, Leigh Thorne, Roy Jackman, Garry C. Whitelam, and Kevin C. Gough*
ADAS UK, Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH. UK.; Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH. UK.; Veterinary Laboratories Agency, Woodham Lane, New Haw, Addlestone, Surrey. KT15 3NB. UK

* To whom correspondence should be addressed. Email: kcg3@le.ac.uk.

Abstract


Disease-associated PrP fragments produced upon in vitro or in vivo proteolysis can provide significant insight into the causal strain of prion disease. A novel molecular strain typing assay is described that used thermolysin digestion of caudal medulla samples to produce PrPres signatures on Western blots that readily distinguished experimental ovine BSE from classical scrapie. Furthermore, the accumulation of such PrPres species within the cerebellum also appeared to be dependent upon TSE strain, allowing the discrimination between two experimental strains of scrapie and the grouping of natural scrapie isolates into two profiles. The occurrence of endogenously produced PrP fragments, namely glycosylated and unglycosylated C2, is also described within different CNS regions; the first detailed description of such scrapie-associated fragments within a natural host. The advent of C2 fragments within defined CNS regions, as compared between BSE and scrapie cases and between two experimental scrapie strains, appeared to be largely dependent upon TSE strain. The combined analyses of C2 fragments and thermolysin-resistant PrP species within caudal medulla, cerebellum and spinal cord samples allowed natural scrapie isolates to be separated into 4 distinct molecular profiles: most isolates produced C2 and PrPres in all CNS regions, a second group lacked detectable cerebellar C2 fragments, one isolate is described that lacked both cerebellar PrPres and C2, and a further isolate lacked detectable C2 within all three CNS regions and also lacked cerebellar PrPres. This CNS region specific deposition of disease-associated PrP species may reflect the natural heterogeneity of scrapie strains within the UK sheep flock.


http://jvi.asm.org/cgi/content/abstract/JVI.00640-07v1?papetoc


TSS



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