Bovine PrP expression levels in transgenic mice influence transmission characteristics of atypical bovine spongiform encephalopathy

Until recently, transmissible spongiform encephalopathy (TSE) disease in cattle was thought to be caused by a single agent strain, bovine spongiform encephalopathy (BSE) (classical BSE or BSE-C). However, due to the initiation of a large-scale surveillance programme throughout Europe, two atypical BSE strains, bovine amyloidotic spongiform encephalopathy (BASE, also named BSE-L) and BSE-H have since been discovered. These atypical BSE isolates have been previously transmitted to a range of transgenic mouse models overexpressing PrP from different species at different levels, on a variety of genetic backgrounds. To control for genetic background and expression level in the analysis of these isolates, we performed here a comprehensive comparison of the neuropathological and molecular properties of all three BSE agents (BASE, BSE-C and BSE-H) upon transmission into the same gene-targeted transgenic mouse line expressing the bovine prion protein (Bov6) and a wild-type control of the same genetic background. Significantly, upon challenge with these BSE agents, we found that BASE did not produce shorter survival times in these mice compared with BSE-C, contrary to previous studies using overexpressing bovine transgenic mice. Amyloid plaques were only present in mice challenged with atypical BSE and neuropathological features, including intensity of PrP deposition in the brain and severity of vacuolar degeneration were less pronounced in BASE compared with BSE-C-challenged mice.

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Presence of subclinical infection in gene-targeted human prion protein transgenic mice exposed to atypical bovine spongiform encephalopathy

Journal of General Virology ◽

10.1099/vir.0.052738-0 ◽

2013 ◽

Vol 94 (12) ◽

pp. 2819-2827 ◽

Cited By ~ 11

Author(s):

Rona Wilson ◽

Karen Dobie ◽

Nora Hunter ◽

Cristina Casalone ◽

Thierry Baron ◽

...

Keyword(s):

Human Health ◽

Bovine Spongiform Encephalopathy ◽

Prion Protein ◽

Disease Transmission ◽

Large Scale ◽

Single Agent ◽

Diagnostic Methods ◽

Transmissible Spongiform Encephalopathies ◽

Spongiform Encephalopathy ◽

Human Prion Protein

The transmission of bovine spongiform encephalopathy (BSE) to humans, leading to variant Creutzfeldt–Jakob disease has demonstrated that cattle transmissible spongiform encephalopathies (TSEs) can pose a risk to human health. Until recently, TSE disease in cattle was thought to be caused by a single agent strain, BSE, also known as classical BSE, or BSE-C. However, due to the initiation of a large-scale surveillance programme throughout Europe, two atypical BSE strains, bovine amyloidotic spongiform encephalopathy (BASE, also named BSE-L) and BSE-H have since been discovered. To model the risk to human health, we previously inoculated these two forms of atypical BSE (BASE and BSE-H) into gene-targeted transgenic (Tg) mice expressing the human prion protein (PrP) (HuTg) but were unable to detect any signs of TSE pathology in these mice. However, despite the absence of TSE pathology, upon subpassage of some BASE-challenged HuTg mice, a TSE was observed in recipient gene-targeted bovine PrP Tg (Bov6) mice but not in HuTg mice. Disease transmission from apparently healthy individuals indicates the presence of subclinical BASE infection in mice expressing human PrP that cannot be identified by current diagnostic methods. However, due to the lack of transmission to HuTg mice on subpassage, the efficiency of mouse-to-mouse transmission of BASE appears to be low when mice express human rather than bovine PrP.

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Vertical Transmission of Bovine Spongiform Encephalopathy Prions Evaluated in a Transgenic Mouse Model

Journal of Virology ◽

10.1128/jvi.79.13.8665-8668.2005 ◽

2005 ◽

Vol 79 (13) ◽

pp. 8665-8668 ◽

Cited By ~ 20

Author(s):

J. Castilla ◽

A. Brun ◽

F. Díaz-San Segundo ◽

F. J. Salguero ◽

A. Gutiérrez-Adán ◽

...

Keyword(s):

Mouse Model ◽

Bovine Spongiform Encephalopathy ◽

Transgenic Mouse ◽

Clinical Stage ◽

Epidemiological Data ◽

Spongiform Encephalopathy ◽

Peripheral Tissues ◽

Transgenic Mouse Line ◽

Intracerebral Administration ◽

Show Evidence

ABSTRACT In this work we show evidence of mother-to-offspring transmission in a transgenic mouse line expressing bovine PrP (boTg) experimentally infected by intracerebral administration of bovine spongiform encephalopathy (BSE) prions. PrPres was detected in brains of newborns from infected mothers only when mating was allowed near to the clinical stage of disease, when brain PrPres deposition could be detected by Western blot analysis. Attempts to detect infectivity in milk after intracerebral inoculation in boTg mice were unsuccessful, suggesting the involvement of other tissues as carriers of prion dissemination. The results shown here prove the ability of BSE prions to spread centrifugally from the central nervous system to peripheral tissues and to offspring in a mouse model. Also, these results may complement previous epidemiological data supporting the occurrence of vertical BSE transmission in cattle.

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Transmission and characterization of bovine spongiform encephalopathy sources in two ovine transgenic mouse lines (TgOvPrP4 and TgOvPrP59)

Journal of General Virology ◽

10.1099/vir.0.82062-0 ◽

2006 ◽

Vol 87 (12) ◽

pp. 3763-3771 ◽

Cited By ~ 27

Author(s):

C. Cordier ◽

A. Bencsik ◽

S. Philippe ◽

D. Bétemps ◽

F. Ronzon ◽

...

Keyword(s):

Transgenic Mice ◽

Bovine Spongiform Encephalopathy ◽

Transgenic Mouse ◽

Neuron Specific Enolase ◽

Infected Sheep ◽

Transmissible Spongiform Encephalopathies ◽

Spongiform Encephalopathy ◽

Wild Type ◽

Spongiform Encephalopathies ◽

Mouse Lines

Transgenic mice expressing the prion protein (PrP) of species affected by transmissible spongiform encephalopathies (TSEs) have recently been produced to facilitate experimental transmission of these diseases by comparison with wild-type mice. However, whilst wild-type mice have largely been described for the discrimination of different TSE strains, including differentiation of agents involved in bovine spongiform encephalopathy (BSE) and scrapie, this has been only poorly described in transgenic mice. Here, two ovine transgenic mouse lines (TgOvPrP4 and TgOvPrP59), expressing the ovine PrP (A136 R154 Q171) under control of the neuron-specific enolase promoter, were studied; they were challenged with brainstem or spinal cord from experimentally BSE-infected sheep (AA136 RR154 QQ171 and AA136 RR154 RR171 genotypes) or brainstem from cattle BSE and natural sheep scrapie. The disease was transmitted successfully from all of these sources, with a mean of approximately 300 days survival following challenge with material from two ARQ-homozygous BSE-infected sheep in TgOvPrP4 mice, whereas the survival period in mice challenged with material from the ARR-homozygous BSE-infected sheep was 423 days on average. It was shown that, in the two ovine transgenic mouse lines, the Western blot characteristics of protease-resistant PrP (PrPres) were similar, whatever the BSE source, with a low apparent molecular mass of the unglycosylated glycoform, a poor labelling by P4 monoclonal antibody and high proportions of the diglycosylated form. With all BSE sources, but not with scrapie, florid plaques were observed in the brains of mice from both transgenic lines. These data reinforce the potential of this recently developed experimental model for the discrimination of BSE from scrapie agents.

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Molecular Analysis of the Protease-Resistant Prion Protein in Scrapie and Bovine Spongiform Encephalopathy Transmitted to Ovine Transgenic and Wild-Type Mice

Journal of Virology ◽

10.1128/jvi.78.12.6243-6251.2004 ◽

2004 ◽

Vol 78 (12) ◽

pp. 6243-6251 ◽

Cited By ~ 47

Author(s):

Thierry Baron ◽

Carole Crozet ◽

Anne-Gaëlle Biacabe ◽

Sandrine Philippe ◽

Jérémie Verchere ◽

...

Keyword(s):

Transgenic Mice ◽

Bovine Spongiform Encephalopathy ◽

Prion Protein ◽

Transmissible Spongiform Encephalopathy ◽

Spongiform Encephalopathy ◽

Infectious Agents ◽

Wild Type ◽

Molecular Features ◽

Different Strains ◽

Scrapie Strains

ABSTRACT The existence of different strains of infectious agents involved in scrapie, a transmissible spongiform encephalopathy (TSE) of sheep and goats, remains poorly explained. These strains can, however, be differentiated by characteristics of the disease in mice and also by the molecular features of the protease-resistant prion protein (PrPres) that accumulates into the infected tissues. For further analysis, we first transmitted the disease from brain samples of TSE-infected sheep to ovine transgenic [Tg(OvPrP4)] and to wild-type (C57BL/6) mice. We show that, as in sheep, molecular differences of PrPres detected by Western blotting can differentiate, in both ovine transgenic and wild-type mice, infection by the bovine spongiform encephalopathy (BSE) agent from most scrapie sources. Similarities of an experimental scrapie isolate (CH1641) with BSE were also likewise found following transmission in ovine transgenic mice. Secondly, we transmitted the disease to ovine transgenic mice by inoculation of brain samples of wild-type mice infected with different experimental scrapie strains (C506M3, 87V, 79A, and Chandler) or with BSE. Features of these strains in ovine transgenic mice were reminiscent of those previously described for wild-type mice, by both ratios and by molecular masses of the different PrPres glycoforms. Moreover, these studies revealed the diversity of scrapie strains and their differences with BSE according to labeling by a monoclonal antibody (P4). These data, in an experimental model expressing the prion protein of the host of natural scrapie, further suggest a genuine diversity of TSE infectious agents and emphasize its linkage to the molecular features of the abnormal prion protein.

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Markedly Increased Susceptibility to Natural Sheep Scrapie of Transgenic Mice Expressing Ovine PrP

Journal of Virology ◽

10.1128/jvi.75.13.5977-5984.2001 ◽

2001 ◽

Vol 75 (13) ◽

pp. 5977-5984 ◽

Cited By ~ 135

Author(s):

Jean-Luc Vilotte ◽

Solange Soulier ◽

Rachid Essalmani ◽

Marie-George Stinnakre ◽

Daniel Vaiman ◽

...

Keyword(s):

Transgenic Mice ◽

Survival Time ◽

Transmissible Spongiform Encephalopathy ◽

Experimental Studies ◽

State Level ◽

Spongiform Encephalopathy ◽

Survival Times ◽

New Host ◽

Improved Model ◽

Sheep Scrapie

ABSTRACT The susceptibility of sheep to scrapie is known to involve, as a major determinant, the nature of the prion protein (PrP) allele, with the VRQ allele conferring the highest susceptibility to the disease. Transgenic mice expressing in their brains three different ovine PrPVRQ-encoding transgenes under an endogenous PrP-deficient genetic background were established. Nine transgenic (tgOv) lines were selected and challenged with two scrapie field isolates derived from VRQ-homozygous affected sheep. All inoculated mice developed neurological signs associated with a transmissible spongiform encephalopathy (TSE) disease and accumulated a protease-resistant form of PrP (PrPres) in their brains. The incubation duration appeared to be inversely related to the PrP steady-state level in the brain, irrespective of the transgene construct. The survival time for animals from the line expressing the highest level of PrP was reduced by at least 1 year compared to those of two groups of conventional mice. With one isolate, the duration of incubation was as short as 2 months, which is comparable to that observed for the rodent TSE models with the briefest survival times. No survival time reduction was observed upon subpassaging of either isolate, suggesting no need for adaptation of the agent to its new host. Overexpression of the transgene was found not to be required for transmission to be accelerated compared to that observed with wild-type mice. Conversely, transgenic mice overexpressing murine PrP were found to be less susceptible than tgOv lines expressing ovine PrP at physiological levels. These data argue that ovine PrPVRQ provided a better substrate for sheep prion replication than did mouse PrP. Altogether, these tgOv mice could be an improved model for experimental studies on natural sheep scrapie.

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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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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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