Transmission of scrapie and sheep-passaged bovine spongiform encephalopathy prions to transgenic mice expressing elk prion protein
Gültekin Tamgüney1,2, Michael W. Miller3, Kurt Giles1,2, Azucena Lemus4, David V. Glidden5, Stephen J. DeArmond1,4 and Stanley B. Prusiner1,2
1 Institute for Neurodegenerative Diseases, University of California, San Francisco, CA 94143-0518, USA 2 Department of Neurology, University of California, San Francisco, CA, USA 3 Colorado Division of Wildlife, Wildlife Research Center, Fort Collins, CO, USA 4 Department of Pathology, University of California, San Francisco, CA, USA 5 Department of Epidemiology and Biostatistics, University of California, San Francisco, CA, USA
Correspondence Stanley B. Prusiner mhtml:%7B33B38F65-8D2E-434D-8F9B-8BDCD77D3066%7Dmid://00000131/!x-usc:mailto:firstname.lastname@example.org
Chronic wasting disease (CWD) is a transmissible, fatal prion disease of cervids and is largely confined to North America. The origin of CWD continues to pose a conundrum: does the disease arise spontaneously or result from some other naturally occurring reservoir? To address whether prions from sheep might be able to cause disease in cervids, we inoculated mice expressing the elk prion protein (PrP) transgene [Tg(ElkPrP) mice] with two scrapie prion isolates. The SSBP/1 scrapie isolate transmitted disease to Tg(ElkPrP) mice with a median incubation time of 270 days, but a second isolate failed to produce neurological dysfunction in these mice. Although prions from cattle with bovine spongiform encephalopathy (BSE) did not transmit to the Tg(ElkPrP) mice, they did transmit after being passaged through sheep. In Tg(ElkPrP) mice, the sheep-passaged BSE prions exhibited an incubation time of approximately 300 days. SSBP/1 prions produced abundant deposits of the disease-causing PrP isoform, denoted PrPSc, in the cerebellum and pons of Tg(ElkPrP) mice, whereas PrPSc accumulation in Tg mice inoculated with sheep-passaged BSE prions was confined to the deep cerebellar nuclei, habenula and the brainstem. The susceptibility of ‘cervidized’ mice to ‘ovinized’ prions raises the question about why CWD has not been reported in other parts of the world where cervids and scrapie-infected sheep coexist.
Supplementary figures and tables are available with the online version of this paper.
Supplementary Fig. S1. Densitometric analysis of glycosylation profiles shows that brain homogenates of diseased Tg mice give rise to PK-resistant PrPSc specific for the expressed transgene and are similar for RML, SSBP/1 and Elk1 prions. Upon second passage in Tg(ElkPrP+/+)12584 mice, all profiles resemble those of cervid prions. Filled bars, diglycosylated bands; shaded bars, monoglycosylated bands; empty bars, unglycosylated bands.
Supplementary Fig. S2. Focal hippocampal vacuolation in the proximal portion of the CA1 region during the first passage of RML prions into Tg(ElkPrP+/0)12577 mice (A–D) and diffuse hippocampal vacuolation during serial passage of RML prions in wild-type FVB mice (E–H). (A, E) Haematoxylin and eosin stain; (B, F) GFAP immunostain; (C, D) immunohistochemical stain of formalin-fixed, paraffin-embedded tissue sections for PrPSc. (G, H) Histoblots of coronal sections of RML-infected, FVB mice immunostained for protease-resistant PrPSc. (G) Coronal section through the dorsal hippocampus and thalamus. (H) Coronal section through the cerebellum and inferior colliculus level of the midbrain. Bars, 50 ìm (A, E); 100 ìm (B, F); 25 ìm (C, D).
Supplementary Table S1. Uninoculated transgenic mice expressing cervid or ovine PrP do not develop spontaneous disease
Transgenic line PrP expression (n-fold) Age (days)* No. ill animals/ no. observed animals† Tg(ElkPrP+/0)12577‡ 2 618 0/8 Tg(ElkPrP+/+)12584 6§ 630 0/7 Tg(OvPrP+/0)14882 0.7 602 0/5 Tg(OvPrP+/+)14882 1.4 601 0/8 *Age of youngest animal when experiment was terminated. †Excludes animals that died from intercurrent disease. ‡Data from Tamgüney et al. (2006). §Based on 3-fold expression for Tg(ElkPrP+/0)12584 mice (Tamgüney et al., 2006). Reference Tamgüney, G., Giles, K., Bouzamondo-Bernstein, E., Bosque, P. J., Miller, M. W., Safar, J., DeArmond, S. J. & Prusiner, S. B. (2006). Transmission of elk and deer prions to transgenic mice. J Virol 80, 9104–9114. Medline Supplementary Table S2. Conformational stability measurements of prion isolates Prion isolates [GdnHCl]½±SD (M) n* SSBP/1 2.1±0.1 2 SSBP/1 ? Tg(ElkPrP+/0)12577 ? Tg(ElkPrP+/+)12584 2.2±0.1 2 SSBP/1 ? Tg(OvPrP+/+)14882 1.6±0.2 3 SSBP/1 ? Tg(ElkPrP+/0)12577 ? Tg(OvPrP+/+)14882 1.7±0.1 2 RML 1.8 1 RML ? Tg(ElkPrP+/0)12577 ? Tg(ElkPrP+/+)12584 1.1±0.1 2 Scrapie 027 2.9±0.1 2 Elk1 2.3 2 Elk1 ? Tg(ElkPrP+/+)12584 ? Tg(ElkPrP+/+)12584 2.1±0.2 2 SA04 ? Tg(ElkPrP+/+)12584 1.4±0.1 2 *Number of assays per isolate or number of brains assayed for passaged inocula.
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REMEMBER, THE NOR-98 ATYPICAL SCRAPIE, WYOMING IS ONE STATE WHERE IT HAS BEEN DOCUMENTED. TO date, i believe that 6 cases of the NOR-98 have been documented to date in the USA. ...tss
Aspects of the Cerebellar Neuropathology in Nor98
Gavier-Widén, D1; Benestad, SL2; Ottander, L1; Westergren, E1 1National Veterinary Insitute, Sweden; 2National Veterinary Institute,
Norway Nor98 is a prion disease of old sheep and goats. This atypical form of scrapie was first described in Norway in 1998. Several features of Nor98 were shown to be different from classical scrapie including the distribution of disease associated prion protein (PrPd) accumulation in the brain. The cerebellum is generally the most affected brain area in Nor98. The study here presented aimed at adding information on the neuropathology in the cerebellum of Nor98 naturally affected sheep of various genotypes in Sweden and Norway. A panel of histochemical and immunohistochemical (IHC) stainings such as IHC for PrPd, synaptophysin, glial fibrillary acidic protein, amyloid, and cell markers for phagocytic cells were conducted. The type of histological lesions and tissue reactions were evaluated. The types of PrPd deposition were characterized. The cerebellar cortex was regularly affected, even though there was a variation in the severity of the lesions from case to case. Neuropil vacuolation was more marked in the molecular layer, but affected also the granular cell layer. There was a loss of granule cells. Punctate deposition of PrPd was characteristic. It was morphologically and in distribution identical with that of synaptophysin, suggesting that PrPd accumulates in the synaptic structures. PrPd was also observed in the granule cell layer and in the white matter. The pathology features of Nor98 in the cerebellum of the affected sheep showed similarities with those of sporadic Creutzfeldt-Jakob disease in humans.
***The pathology features of Nor98 in the cerebellum of the affected sheep showed similarities with those of sporadic Creutzfeldt-Jakob disease in humans.
Monday, December 1, 2008 When Atypical Scrapie cross species barriers
Research Project: Strain Typing of Chronic Wasting Disease (Cwd) and Scrapie by Intracerebral Inoculation into Transgenic and Inbred Mouse Lines
Location: Animal Diseases Research
2007 Annual Report
1a.Objectives (from AD-416)
To identify and differentiate typical and atypical case samples of CWD and Scrapie by characterizing the biologic phenotype and pathologic profile of these agents when administered to susceptible lines of transgenic and inbred mice.
1b.Approach (from AD-416)
Tissue samples from deer, elk, sheep and goats with Transmissible Spongiform Encephalopathy (TSE) will be administered to mice by intracerebral injection. Multiple tissue types will be included, such as samples of brain, lymph node, blood, urine, feces, antler velvet and muscle. Mouse models used as recipient hosts will include both preexisting and recently created transgenic and inbred mouse lines. Recipient mouse phenotype will be evaluated by measuring clinical response, population disease rate, incubation time, and pathologic profile within the central nervous system (CNS). Pathologic profile of CNS lesion foci is assessed by evaluating anatomic localization, spongiform change, astrocytic gliosis, and deposition of protease resistant prion protein. BSL-1; 9-4-06. Documents SCA with U. of WA.
This report serves to document research conducted under a specific cooperative agreement between ARS and the University of Washington. Additional details of research can be found in the report for the parent project 5348-32000-026-00D Transmissible Spongiform Encephalopathies: the role of genetics, strain variation, and environmental contamination in disease control. The purpose of this new SCA is to identify and differentiate typical and atypical case samples of CWD and scrapie by characterizing the biologic phenotype and pathologic profile of these agents when administered to susceptible lines of transgenic and inbred mice. There will be weekly interactions between the ADODR, ADRU scientists and University of Washington collaborators through personnel visits, conference calls and emails.
Chronic Wasting Disease and Potential Transmission to Humans
Ermias D. Belay,* Ryan A. Maddox,* Elizabeth S. Williams,? Michael W. Miller,? Pierluigi Gambetti,§ and Lawrence B. Schonberger*
*Centers for Disease Control and Prevention, Atlanta, Georgia, USA; ?University of Wyoming, Laramie, Wyoming, USA; ?Colorado Division of Wildlife, Fort Collins, Colorado, USA; and §Case Western Reserve University, Cleveland, Ohio, USA
Suggested citation for this article: Belay ED, Maddox RA, Williams ES, Miller MW, Gambetti P, Schonberger LB. Chronic wasting disease and potential transmission to humans. Emerg Infect Dis [serial on the Internet]. 2004 Jun [date cited]. Available from:http://www.cdc.gov/ncidod/EID/vol10no6/03-1082.htm
Chronic wasting disease (CWD) of deer and elk is endemic in a tri-corner area of Colorado, Wyoming, and Nebraska, and new foci of CWD have been detected in other parts of the United States. Although detection in some areas may be related to increased surveillance, introduction of CWD due to translocation or natural migration of animals may account for some new foci of infection. Increasing spread of CWD has raised concerns about the potential for increasing human exposure to the CWD agent. The foodborne transmission of bovine spongiform encephalopathy to humans indicates that the species barrier may not completely protect humans from animal prion diseases. Conversion of human prion protein by CWD-associated prions has been demonstrated in an in vitro cell-free experiment, but limited investigations have not identified strong evidence for CWD transmission to humans. More epidemiologic and laboratory studies are needed to monitor the possibility of such transmissions.
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Volume 12, Number 10-October 2006
Human Prion Disease and Relative Risk Associated with Chronic Wasting Disease
Samantha MaWhinney,* W. John Pape,? Jeri E. Forster,* C. Alan Anderson,?§ Patrick Bosque,?¶ and Michael W. Miller#
*University of Colorado at Denver and Health Sciences Center, Denver, Colorado, USA; ?Colorado Department of Public Health and Environment, Denver, Colorado, USA; ?University of Colorado School of Medicine, Denver, Colorado, USA; §Denver Veteran's Affairs Medical Center, Denver, Colorado, USA; ¶Denver Health Medical Center, Denver, Colorado, USA; and #Colorado Division of Wildlife, Fort Collins, Colorado, USA
Suggested citation for this article
The transmission of the prion disease bovine spongiform encephalopathy (BSE) to humans raises concern about chronic wasting disease (CWD), a prion disease of deer and elk. In 7 Colorado counties with high CWD prevalence, 75% of state hunting licenses are issued locally, which suggests that residents consume most regionally harvested game. We used Colorado death certificate data from 1979 through 2001 to evaluate rates of death from the human prion disease Creutzfeldt-Jakob disease (CJD). The relative risk (RR) of CJD for CWD-endemic county residents was not significantly increased (RR 0.81, 95% confidence interval [CI] 0.40-1.63), and the rate of CJD did not increase over time (5-year RR 0.92, 95% CI 0.73-1.16). In Colorado, human prion disease resulting from CWD exposure is rare or nonexistent. However, given uncertainties about the incubation period, exposure, and clinical presentation, the possibility that the CWD agent might cause human disease cannot be eliminated.
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