Large‐scale prion protein genotyping in Canadian caribou populations and potential impact on chronic wasting disease susceptibility

Chronic wasting disease (CWD) is a prion-mediated, transmissible disease of cervids, including deer ( Odocoileus spp.), which is characterized by spongiform encephalopathy and death of the prion-infected animals. Official surveillance in the United States using immunohistochemistry (IHC) and ELISA entails the laborious collection of lymphoid and/or brainstem tissue after death. New, highly sensitive prion detection methods, such as real-time quaking-induced conversion (RT-QuIC), have shown promise in detecting abnormal prions from both antemortem and postmortem specimens. We compared RT-QuIC with ELISA and IHC for CWD detection utilizing deer retropharyngeal lymph node (RLN) tissues in a diagnostic laboratory setting. The RLNs were collected postmortem from hunter-harvested animals. RT-QuIC showed 100% sensitivity and specificity for 50 deer RLN (35 positive by both IHC and ELISA, 15 negative) included in our study. All deer were also genotyped for PRNP polymorphism. Most deer were homozygous at codons 95, 96, 116, and 226 (QQ/GG/AA/QQ genotype, with frequency 0.86), which are the codons implicated in disease susceptibility. Heterozygosity was noticed in Pennsylvania deer, albeit at a very low frequency, for codons 95GS (0.06) and 96QH (0.08), but deer with these genotypes were still found to be CWD prion-infected.

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Efficient In Vitro Amplification of Chronic Wasting Disease PrPRES

Journal of Virology ◽

10.1128/jvi.00635-07 ◽

2007 ◽

Vol 81 (17) ◽

pp. 9605-9608 ◽

Cited By ~ 71

Author(s):

Timothy D. Kurt ◽

Matthew R. Perrott ◽

Carol J. Wilusz ◽

Jeffrey Wilusz ◽

Surachai Supattapone ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Body Fluids ◽

The Body ◽

Wasting Disease ◽

Highly Efficient ◽

Significant Step

ABSTRACT Chronic wasting disease (CWD) of cervids is associated with conversion of the normal cervid prion protein, PrPC, to a protease-resistant conformer, PrPCWD. Here we report the use of both nondenaturing amplification and protein-misfolding cyclic amplification (PMCA) to amplify PrPCWD in vitro. Normal brains from deer, transgenic mice expressing cervid PrPC [Tg(cerPrP)1536 mice], and ferrets supported amplification. PMCA using normal Tg(cerPrP)1536 brains as the PrPC substrate produced >6.5 × 109-fold amplification after six rounds. Highly efficient in vitro amplification of PrPCWD is a significant step toward detection of PrPCWD in the body fluids or excreta of CWD-susceptible species.

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Survey of Cattle in Northeast Colorado for Evidence of Chronic Wasting Disease: Geographical and High-Risk Targeted Sample

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870301500309 ◽

2003 ◽

Vol 15 (3) ◽

pp. 274-277 ◽

Cited By ~ 10

Author(s):

Daniel H. Gould ◽

James L. Voss ◽

Michael W. Miller ◽

Annette M. Bachand ◽

Bruce A. Cummings ◽

...

Keyword(s):

High Risk ◽

Large Scale ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

Proteinase K ◽

Spongiform Encephalopathy ◽

Wasting Disease ◽

Adult Cattle ◽

Endemic Areas ◽

Geographically Targeted

A geographically targeted survey of potentially high-risk, adult cattle in chronic wasting disease (CWD)–endemic areas in Colorado was initiated to assess the possibility of the spread of CWD from deer to cattle under natural conditions. Surveyed cattle were sympatric with free-roaming deer in geographically defined areas where CWD occurs and where CWD prevalence has been estimated. To qualify for inclusion in the survey, cattle had to be at least 4 years old and had to have spent a minimum of 4 years in surveyed areas. Brains from culled cattle were examined microscopically and immunohistochemically for tissue alterations indicative of a transmissible spongiform encephalopathy (TSE). Two hundred sixty-two brains were suitable for evaluation and were found to lack changes indicative of a TSE infection. Prion deposition was not demonstrable using a method involving formic acid and proteinase-K treatment before application of monoclonal antibody to bovine prion protein (F99/97.6.1). Some incidental neuropathologic changes unrelated to those of TSEs were detected. Findings from this study suggest that large-scale spread of CWD from deer to cattle under natural range conditions in CWD-endemic areas of northeast Colorado is unlikely.

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Recombinant prion protein vaccination of transgenic elk PrP mice and reindeer overcomes self-tolerance and protects mice against chronic wasting disease

Journal of Biological Chemistry ◽

10.1074/jbc.ra118.004810 ◽

2018 ◽

Vol 293 (51) ◽

pp. 19812-19822 ◽

Cited By ~ 3

Author(s):

Dalia H. Abdelaziz ◽

Simrika Thapa ◽

Jenna Brandon ◽

Justine Maybee ◽

Lauren Vankuppeveld ◽

...

Keyword(s):

Prion Protein ◽

Chronic Wasting Disease ◽

Wasting Disease ◽

Self Tolerance ◽

Recombinant Prion Protein

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Species barriers for chronic wasting disease by in vitro conversion of prion protein

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2007.10.087 ◽

2007 ◽

Vol 364 (4) ◽

pp. 796-800 ◽

Cited By ~ 16

Author(s):

Li Li ◽

Michael B. Coulthart ◽

Aru Balachandran ◽

Avi Chakrabartty ◽

Neil R. Cashman

Keyword(s):

Prion Protein ◽

Chronic Wasting Disease ◽

Wasting Disease

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Detection of prion protein in the cerebrospinal fluid of elk (Cervus canadensis nelsoni) with chronic wasting disease using protein misfolding cyclic amplification

Journal of Veterinary Diagnostic Investigation ◽

10.1177/1040638712448060 ◽

2012 ◽

Vol 24 (4) ◽

pp. 746-749 ◽

Cited By ~ 13

Author(s):

Tracy A. Nichols ◽

Terry R. Spraker ◽

Tom Gidlewski ◽

Jenny G. Powers ◽

Glenn C. Telling ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Prion Protein ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Wasting Disease ◽

Protein Misfolding Cyclic Amplification ◽

Cervus Canadensis

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Experimental Transmission of the Chronic Wasting Disease Agent to Swine after Oral or Intracranial Inoculation

Journal of Virology ◽

10.1128/jvi.00926-17 ◽

2017 ◽

Vol 91 (19) ◽

Cited By ~ 14

Author(s):

S. Jo Moore ◽

M. Heather West Greenlee ◽

Naveen Kondru ◽

Sireesha Manne ◽

Jodi D. Smith ◽

...

Keyword(s):

Prion Protein ◽

Prion Disease ◽

Chronic Wasting Disease ◽

Mouse Bioassay ◽

Lymphoid Tissues ◽

Species Barrier ◽

Wasting Disease ◽

Disease Agent ◽

Domestic Swine

ABSTRACT Chronic wasting disease (CWD) is a naturally occurring, fatal neurodegenerative disease of cervids. The potential for swine to serve as hosts for the agent of CWD is unknown. The purpose of this study was to investigate the susceptibility of swine to the CWD agent following experimental oral or intracranial inoculation. Crossbred piglets were assigned to three groups, intracranially inoculated (n = 20), orally inoculated (n = 19), and noninoculated (n = 9). At approximately the age at which commercial pigs reach market weight, half of the pigs in each group were culled (“market weight” groups). The remaining pigs (“aged” groups) were allowed to incubate for up to 73 months postinoculation (mpi). Tissues collected at necropsy were examined for disease-associated prion protein (PrPSc) by Western blotting (WB), antigen capture enzyme immunoassay (EIA), immunohistochemistry (IHC), and in vitro real-time quaking-induced conversion (RT-QuIC). Brain samples from selected pigs were also bioassayed in mice expressing porcine prion protein. Four intracranially inoculated aged pigs and one orally inoculated aged pig were positive by EIA, IHC, and/or WB. By RT-QuIC, PrPSc was detected in lymphoid and/or brain tissue from one or more pigs in each inoculated group. The bioassay was positive in four out of five pigs assayed. This study demonstrates that pigs can support low-level amplification of CWD prions, although the species barrier to CWD infection is relatively high. However, detection of infectivity in orally inoculated pigs with a mouse bioassay raises the possibility that naturally exposed pigs could act as a reservoir of CWD infectivity. IMPORTANCE We challenged domestic swine with the chronic wasting disease agent by inoculation directly into the brain (intracranially) or by oral gavage (orally). Disease-associated prion protein (PrPSc) was detected in brain and lymphoid tissues from intracranially and orally inoculated pigs as early as 8 months of age (6 months postinoculation). Only one pig developed clinical neurologic signs suggestive of prion disease. The amount of PrPSc in the brains and lymphoid tissues of positive pigs was small, especially in orally inoculated pigs. Regardless, positive results obtained with orally inoculated pigs suggest that it may be possible for swine to serve as a reservoir for prion disease under natural conditions.

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