Abstract
Transmissible spongiform encephalopathies (TSEs), including scrapie in sheep (Ovis aries), are fatal neurodegenerative diseases caused by the misfolding of the cellular prion protein (PrPC) into a â-rich conformer (PrPSc) that accumulates into higher-order structures in the brain and other tissues. Distinct strains of TSEs exist, characterized by different pathologic profiles upon passage into rodents and representing distinct conformations of PrPSc. One biochemical method of distinguishing strains is the stability of PrPSc as determined by unfolding in guanidine hydrochloride (GdnHCl), which is tightly and positively correlated with the incubation time of disease upon passage into mice. Here, we utilize a rapid, protease-free version of the stability assay to characterize naturally occurring scrapie samples, including a fast-acting scrapie inoculum for which incubation time is highly dependent on the amino acid at codon 136 of the prion protein. We utilize the stability methodology to identify the presence of two distinct isolates in the inoculum, and compare isolate properties to those of a host-stabilized reference scrapie isolate (NADC 13-7) in order to assess the stability/incubation time correlation in a natural host system. We demonstrate the utility of the stability methodology in characterizing TSE isolates throughout serial passage in livestock, which is applicable to a range of natural host systems, including strains of bovine spongiform encephalopathy and chronic wasting disease.
Highlights
Prion diseases, including bovine spongiform encephalopathy (BSE) in cattle, scrapie in sheep and goats, chronic wasting disease (CWD) in cervids, and Creutzfeldt-Jakob disease (CJD) in humans, are neurodegenerative diseases that manifest as behavioral changes and/or neurological signs and eventually progress to death
Known as transmissible spongiform encephalopathies (TSEs), prion diseases are caused by misfolding of the cellular, protease-sensitive prion protein (PrPC) into an infectious, more protease-resistant form (PrPSc) that accumulates in the nervous system and certain other body tissues
As a model system for examining correlations between incubation time and PrPSc stability, we began by considering the properties of a United States sheep scrapie inoculum that exhibited a strong dependence of host genotype on the disease incubation time
Summary
Prion diseases, including bovine spongiform encephalopathy (BSE) in cattle, scrapie in sheep and goats, chronic wasting disease (CWD) in cervids, and Creutzfeldt-Jakob disease (CJD) in humans, are neurodegenerative diseases that manifest as behavioral changes and/or neurological signs and eventually progress to death. We utilized the ELISA-based stability assay to consider the relationships between PrPSc stability, incubation time, and host genotype in a natural host system for prion disease: sheep (Ovis aries) experimentally infected with classical scrapie.
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