Resistance to Chronic Wasting Disease in Transgenic Mice Expressing a Naturally Occurring Allelic Variant of Deer Prion Protein

ABSTRACT Prion protein (PrP) is a required factor for susceptibility to transmissible spongiform encephalopathy or prion diseases. In transgenic mice, expression of prion protein (PrP) from another species often confers susceptibility to prion disease from that donor species. For example, expression of deer or elk PrP in transgenic mice has induced susceptibility to chronic wasting disease (CWD), the prion disease of cervids. In the current experiments, transgenic mice expressing two naturally occurring allelic variants of deer PrP with either glycine (G) or serine (S) at residue 96 were found to differ in susceptibility to CWD infection. G96 mice were highly susceptible to infection, and disease appeared starting as early as 160 days postinfection. In contrast, S96 mice showed no evidence of disease or generation of disease-associated protease-resistant PrP (PrPres) over a 600-day period. At the time of clinical disease, G96 mice showed typical vacuolar pathology and deposition of PrPres in many brain regions, and in some individuals, extensive neuronal loss and apoptosis were noted in the hippocampus and cerebellum. Extraneural accumulation of PrPres was also noted in spleen and intestinal tissue of clinically ill G96 mice. These results demonstrate the importance of deer PrP polymorphisms in susceptibility to CWD infection. Furthermore, this deer PrP transgenic model is the first to demonstrate extraneural accumulation of PrPres in spleen and intestinal tissue and thus may prove useful in studies of CWD pathogenesis and transmission by oral or other natural routes of infection.

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Transmission and Adaptation of Chronic Wasting Disease to Hamsters and Transgenic Mice: Evidence for Strains

Journal of Virology ◽

10.1128/jvi.02474-06 ◽

2007 ◽

Vol 81 (8) ◽

pp. 4305-4314 ◽

Cited By ~ 72

Author(s):

Gregory J. Raymond ◽

Lynne D. Raymond ◽

Kimberly D. Meade-White ◽

Andrew G. Hughson ◽

Cynthia Favara ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

Wild Type Mouse ◽

Mouse Strains ◽

Spongiform Encephalopathy ◽

Wild Type ◽

Wasting Disease ◽

Incubation Periods

ABSTRACT In vitro screening using the cell-free prion protein conversion system indicated that certain rodents may be susceptible to chronic wasting disease (CWD). Therefore, CWD isolates from mule deer, white-tailed deer, and elk were inoculated intracerebrally into various rodent species to assess the rodents' susceptibility and to develop new rodent models of CWD. The species inoculated were Syrian golden, Djungarian, Chinese, Siberian, and Armenian hamsters, transgenic mice expressing the Syrian golden hamster prion protein, and RML Swiss and C57BL10 wild-type mice. The transgenic mice and the Syrian golden, Chinese, Siberian, and Armenian hamsters had limited susceptibility to certain of the CWD inocula, as evidenced by incomplete attack rates and long incubation periods. For serial passages of CWD isolates in Syrian golden hamsters, incubation periods rapidly stabilized, with isolates having either short (85 to 89 days) or long (408 to 544 days) mean incubation periods and distinct neuropathological patterns. In contrast, wild-type mouse strains and Djungarian hamsters were not susceptible to CWD. These results show that CWD can be transmitted and adapted to some species of rodents and suggest that the cervid-derived CWD inocula may have contained or diverged into at least two distinct transmissible spongiform encephalopathy strains.

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Cell-Based Quantification of Chronic Wasting Disease Prions

Journal of Virology ◽

10.1128/jvi.00633-10 ◽

2010 ◽

Vol 84 (16) ◽

pp. 8322-8326 ◽

Cited By ~ 54

Author(s):

Jifeng Bian ◽

Dana Napier ◽

Vadim Khaychuck ◽

Rachel Angers ◽

Catherine Graham ◽

...

Keyword(s):

Transgenic Mice ◽

Cell Cultures ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

Spongiform Encephalopathy ◽

Wasting Disease ◽

Cell Assay ◽

Naturally Occurring ◽

Experimental Strains ◽

Isolated Cell

ABSTRACT Cell-based measurement of prion infectivity is currently restricted to experimental strains of mouse-adapted scrapie. Having isolated cell cultures with susceptibility to prions from diseased elk, we describe a modification of the scrapie cell assay allowing evaluation of prions causing chronic wasting disease, a naturally occurring transmissible spongiform encephalopathy. We compare this cervid prion cell assay to bioassays in transgenic mice, the only other existing method for quantification, and show this assay to be a relatively economical and expedient alternative that will likely facilitate studies of this important prion disease.

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Humanized Transgenic Mice Are Resistant to Chronic Wasting Disease Prions From Norwegian Reindeer and Moose

The Journal of Infectious Diseases ◽

10.1093/infdis/jiab033 ◽

2021 ◽

Author(s):

Jonathan D F Wadsworth ◽

Susan Joiner ◽

Jacqueline M Linehan ◽

Kezia Jack ◽

Huda Al-Doujaily ◽

...

Keyword(s):

Transgenic Mice ◽

Brain Tissue ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

Spongiform Encephalopathy ◽

Zoonotic Potential ◽

Wasting Disease ◽

Human Prion Protein ◽

Prion Transmission ◽

Wild Reindeer

Abstract Chronic wasting disease (CWD) is the transmissible spongiform encephalopathy or prion disease affecting cervids. In 2016, the first cases of CWD were reported in Europe in Norwegian wild reindeer and moose. The origin and zoonotic potential of these new prion isolates remain unknown. In this study to investigate zoonotic potential we inoculated brain tissue from CWD-infected Norwegian reindeer and moose into transgenic mice overexpressing human prion protein. After prolonged postinoculation survival periods no evidence for prion transmission was seen, suggesting that the zoonotic potential of these isolates is low.

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Inhibition of Protease-Resistant Prion Protein Formation in a Transformed Deer Cell Line Infected with Chronic Wasting Disease

Journal of Virology ◽

10.1128/jvi.80.2.596-604.2006 ◽

2006 ◽

Vol 80 (2) ◽

pp. 596-604 ◽

Cited By ~ 48

Author(s):

Gregory J. Raymond ◽

Emily A. Olsen ◽

Kil Sun Lee ◽

Lynne D. Raymond ◽

P. Kruger Bryant ◽

...

Keyword(s):

Cell Line ◽

Prion Protein ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

Mule Deer ◽

Simian Virus 40 ◽

Spongiform Encephalopathy ◽

Wasting Disease ◽

Neuronal Marker

ABSTRACT Chronic wasting disease (CWD) is an emerging transmissible spongiform encephalopathy (prion disease) of North American cervids, i.e., mule deer, white-tailed deer, and elk (wapiti). To facilitate in vitro studies of CWD, we have developed a transformed deer cell line that is persistently infected with CWD. Primary cultures derived from uninfected mule deer brain tissue were transformed by transfection with a plasmid containing the simian virus 40 genome. A transformed cell line (MDB) was exposed to microsomes prepared from the brainstem of a CWD-affected mule deer. CWD-associated, protease-resistant prion protein (PrPCWD) was used as an indicator of CWD infection. Although no PrPCWD was detected in any of these cultures after two passes, dilution cloning of cells yielded one PrPCWD-positive clone out of 51. This clone, designated MDBCWD, has maintained stable PrPCWD production through 32 serial passes thus far. A second round of dilution cloning yielded 20 PrPCWD-positive subclones out of 30, one of which was designated MDBCWD2. The MDBCWD2 cell line was positive for fibronectin and negative for microtubule-associated protein 2 (a neuronal marker) and glial fibrillary acidic protein (an activated astrocyte marker), consistent with derivation from brain fibroblasts (e.g., meningeal fibroblasts). Two inhibitors of rodent scrapie protease-resistant PrP accumulation, pentosan polysulfate and a porphyrin compound, indium (III) meso-tetra(4-sulfonatophenyl)porphine chloride, potently blocked PrPCWD accumulation in MDBCWD cells. This demonstrates the utility of these cells in a rapid in vitro screening assay for PrPCWD inhibitors and suggests that these compounds have potential to be active against CWD in vivo.

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Comparison of Abnormal Prion Protein Glycoform Patterns from Transmissible Spongiform Encephalopathy Agent-Infected Deer, Elk, Sheep, and Cattle

Journal of Virology ◽

10.1128/jvi.76.23.12365-12368.2002 ◽

2002 ◽

Vol 76 (23) ◽

pp. 12365-12368 ◽

Cited By ~ 37

Author(s):

Richard E. Race ◽

Anne Raines ◽

Thierry G. M. Baron ◽

Michael W. Miller ◽

Allen Jenny ◽

...

Keyword(s):

Bovine Spongiform Encephalopathy ◽

Prion Protein ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

Spongiform Encephalopathy ◽

Wasting Disease

ABSTRACT Analysis of abnormal prion protein glycoform patterns from chronic wasting disease (CWD)-affected deer and elk, scrapie-affected sheep and cattle, and cattle with bovine spongiform encephalopathy failed to identify patterns capable of reliably distinguishing these transmissible spongiform encephalopathy diseases. However, PrP-res patterns sometimes differed among individual animals, suggesting infection by different or multiple CWD strains in some species.

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Generation of human chronic wasting disease in transgenic mice

Acta Neuropathologica Communications ◽

10.1186/s40478-021-01262-y ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

Zerui Wang ◽

Kefeng Qin ◽

Manuel V. Camacho ◽

Ignazio Cali ◽

Jue Yuan ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Disease ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Cellular Prion Protein ◽

Spongiform Encephalopathy ◽

Species Barrier ◽

Wasting Disease ◽

Bona Fide

AbstractChronic wasting disease (CWD) is a cervid prion disease caused by the accumulation of an infectious misfolded conformer (PrPSc) of cellular prion protein (PrPC). It has been spreading rapidly in North America and also found in Asia and Europe. Although bovine spongiform encephalopathy (i.e. mad cow disease) is the only animal prion disease known to be zoonotic, the transmissibility of CWD to humans remains uncertain. Here we report the generation of the first CWD-derived infectious human PrPSc by elk CWD PrPSc-seeded conversion of PrPC in normal human brain homogenates using in vitro protein misfolding cyclic amplification (PMCA). Western blotting with human PrP selective antibody confirmed that the PMCA-generated protease-resistant PrPSc was derived from the human PrPC substrate. Two lines of humanized transgenic mice expressing human PrP with either Val or Met at the polymorphic codon 129 developed clinical prion disease following intracerebral inoculation with the PMCA-generated CWD-derived human PrPSc. Diseased mice exhibited distinct PrPSc patterns and neuropathological changes in the brain. Our study, using PMCA and animal bioassays, provides the first evidence that CWD PrPSc can cross the species barrier to convert human PrPC into infectious PrPSc that can produce bona fide prion disease when inoculated into humanized transgenic mice.

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Comparison of Histological Lesions and Immunohistochemical Staining of Proteinase-resistant Prion Protein in a Naturally Occurring Spongiform Encephalopathy of Free-ranging Mule Deer (Odocoileus hemionus) with Those of Chronic Wasting Disease of Captive Mule Deer

Veterinary Pathology ◽

10.1354/vp.39-1-110 ◽

2002 ◽

Vol 39 (1) ◽

pp. 110-119 ◽

Cited By ~ 66

Author(s):

T. R. Spraker ◽

R. R. Zink ◽

B. A. Cummings ◽

M. A. Wild ◽

M. W. Miller ◽

...

Keyword(s):

Prion Protein ◽

Immunohistochemical Staining ◽

Chronic Wasting Disease ◽

Mule Deer ◽

Odocoileus Hemionus ◽

Spongiform Encephalopathy ◽

Wasting Disease ◽

Naturally Occurring ◽

Free Ranging

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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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