Proteinase K enhanced immunoreactivity of the prion protein-specific monoclonal antibody 2A11

Brun, Alejandro; Castilla, Joaquı́n; Ramírez, Miguel Ángel; Prager, Kai; Parra, Beatriz; Salguero, Francisco J.; Shiveral, Diane; Sánchez, Carmen; Sánchez‐Vizcaíno, José Manuel; Douglas, Arthur V.; Torres, Juan María

doi:10.1016/j.neures.2003.09.004

Cited by 32 publications

(27 citation statements)

References 31 publications

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“…In original BSE 5 , BSE/sheep, or BoPrP-Tg110 mice inoculated with BSE 5 or BSE/sheep, no differences could be observed in terms of PrP res (i) electrophoresis mobility (Fig. 4A, lanes 1 to 6) (ii) glycoprofile (Fig.…”

Section: Resultsmentioning

confidence: 94%

Sheep-Passaged Bovine Spongiform Encephalopathy Agent Exhibits Altered Pathobiological Properties in Bovine-PrP Transgenic Mice

Espinosa

Andréoletti

Castilla

et al. 2007

J Virol

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Sheep can be experimentally infected with bovine spongiform encephalopathy (BSE), and the ensuing disease is similar to scrapie in terms of pathogenesis and clinical signs. BSE infection in sheep is an animal and human health concern. In this study, the transmission in BoPrP-Tg110 mice of prions from BSE-infected sheep was examined and compared to the transmission of original cattle BSE in cattle and sheep scrapie prions. Our results indicate no transmission barrier for sheep BSE prions to infect BoPrP-Tg110 mice, but the course of the disease is accelerated compared to the effects of the original BSE isolate. The shortened incubation period of sheep BSE in the model was conserved in subsequent passage in BoPrP-Tg110 mice, indicating that it is not related to infectious titer differences. Biochemical signature, lesion profile, and PrP Sc deposition pattern of both cattle and sheep BSE were similar. In contrast, all three sheep scrapie isolates tested showed an evident transmission barrier and further adaptation in subsequent passage. Taken together, those data indicate that BSE agent can be altered by crossing a species barrier, raising concerns about the virulence of this new prion towards other species, including humans. The BoPrP-Tg110 mouse bioassay should be considered as a valuable tool for discriminating scrapie and BSE in sheep.

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Section: Resultsmentioning

confidence: 94%

Sheep-Passaged Bovine Spongiform Encephalopathy Agent Exhibits Altered Pathobiological Properties in Bovine-PrP Transgenic Mice

Espinosa

Andréoletti

Castilla

et al. 2007

J Virol

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“…Samples were precleared by centrifugation at 2,000 ϫ g for 5 min, after which an equal volume of 2ϫ SDS reducing sample loading buffer was added to all the samples, and each one was boiled for 5 min before loading on an SDS-12% polyacrylamide gel. The expression levels of vole PrP C in brains from the mouse lines were analyzed by Western blotting (WB) and compared with the PrP C content in Bv109M brain (data not shown) using monoclonal antibody (MAb) 2A11 (which recognizes the 160 QVYYRPVDQ 168 epitope, numbered according to the vole PrP sequence) as previously described (20). The BvPrP-Tg407 mouse line expressing Bv109M-PrP C (referred to here as BvPrP-Tg407) was selected for further experiments on the basis of PrP C expression.…”

Section: Methodsmentioning

confidence: 99%

“…Transgenic PrP C expression in homozygous BvPrP-Tg407 mice was checked by Western blotting using a specific anti-PrP monoclonal antibody (2A11 [20]). Brain PrP C expression levels for the homozygous mice were found to be similar to the PrP C levels found in bank voles, and therefore, BvPrP-Tg407 was chosen as the only transgenic-mouse line used in the study.…”

Section: Bv109m Prpmentioning

confidence: 99%

PrP ^C Governs Susceptibility to Prion Strains in Bank Vole, While Other Host Factors Modulate Strain Features

Espinosa

Nonno

Bari

et al. 2016

J Virol

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Bank vole is a rodent species that shows differential susceptibility to the experimental transmission of different prion strains. In this work, the transmission features of a panel of diverse prions with distinct origins were assayed both in bank vole expressing methionine at codon 109 (Bv109M) and in transgenic mice expressing physiological levels of bank vole PrP C (the BvPrP-Tg407 mouse line). This work is the first systematic comparison of the transmission features of a collection of prion isolates, representing a panel of diverse prion strains, in a transgenic-mouse model and in its natural counterpart. The results showed very similar transmission properties in both the natural species and the transgenic-mouse model, demonstrating the key role of the PrP amino acid sequence in prion transmission susceptibility. However, differences in the PrP Sc types propagated by Bv109M and BvPrP-Tg407 suggest that host factors other than PrP C modulate prion strain features. IMPORTANCEThe differential susceptibility of bank voles to prion strains can be modeled in transgenic mice, suggesting that this selective susceptibility is controlled by the vole PrP sequence alone rather than by other species-specific factors. Differences in the phenotypes observed after prion transmissions in bank voles and in the transgenic mice suggest that host factors other than the PrP C sequence may affect the selection of the substrain replicating in the animal model. Transmissible spongiform encephalopathies (TSEs), or prion diseases, are a group of fatal neurodegenerative diseases that include Creutzfeldt-Jakob disease (CJD) in humans, bovine spongiform encephalopathy (BSE) in cattle, and scrapie in small ruminants. The prion agent consists mainly, if not entirely, of PrP Sc , which is an abnormal isoform of the host-encoded prion cellular protein, PrP C (1-4). Prion propagation is considered to occur in a two-step process in which aggregates of PrP Sc bind new PrP C monomers and then induce their conformational change into PrP Sc . The conversion into PrP Sc results in changes in physical properties, such as decreased solubility in nondenaturing detergents, an increase in ␤-pleated-sheet content, and increased resistance to proteolytic degradation.The occurrence of different TSE strains has been described in several species. Upon experimental transmission, TSE strains show distinctive incubation periods and neuropathological features that are maintained after subpassage in the same species. These strains correspond to different PrP Sc conformations, according to the prion hypothesis (5). Prion strain transmission among different species seems to be driven by the differences in the PrP amino acid sequences and by the prion strain being transmitted (6). Transgenic-mouse models null for the murine Prnp gene and expressing PrP C from another species (such as bovine, ovine, porcine, or human), have been used in the prion research field as useful tools to characterize prion strains and to learn about the susceptibility of the species t...

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“…Tissue sections were prepared as described previously (Castilla et al, 2003) and then incubated overnight at 4°C with primary 6H4 monoclonal antibody (mAb) (Prionics, Schlieren, Switzerland) or 2A11 mAb (Brun et al, 2004) diluted 1:400 in PBS. The slides were then treated with the secondary antibody and developed according to a previously described procedure (Castilla et al, 2003).…”

Section: Plasmid Constructsmentioning

confidence: 99%

Subclinical Bovine Spongiform Encephalopathy Infection in Transgenic Mice Expressing Porcine Prion Protein

Castilla

Gutiérrez‐Adán²,

Brun³

et al. 2004

J. Neurosci.

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The bovine-porcine species barrier to bovine spongiform encephalopathy (BSE) infection was explored by generating transgenic mouse lines expressing the porcine prion protein (PrP) gene. All of the porcine transgenic (poTg) mice showed clinical signs of BSE after intracerebral inoculation with a high-titer BSE inoculum. The protease-resistant PrP (PrP res ) was detected in 14% (3 of 22) of the BSE-infected poTg mice by immunohistochemical or immunoblot analysis. Despite being able to infect 42% (5 of 12) of control mice, a low-dose BSE inoculum failed to penetrate the species barrier in our poTg mouse model. The findings of these infectivity studies suggest that there is a strong species barrier between cows and pigs. However, after second-passage infection of poTg mice using brain homogenates of BSE-inoculated mice scoring negative for the incoming prion protein as inoculum, it was possible to detect the presence of the infectious agent. Thus, porcine-adapted BSE inocula were efficient at infecting poTg mice, giving rise to an incubation period substantially reduced from 300 to 177 d after inoculation and to the presence of PrP res in 100% (21 of 21) of the mice. We were therefore able to conclude that initial exposure to the bovine prion may lead to subclinical infection such that brain homogenates from poTg mice classified as uninfected on the basis of the absence of PrP res are infectious when used to reinoculate poTg mice. Collectively, our findings suggest that these poTg mice could be used as a sensitive bioassay model for prion detection in pigs.

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Proteinase K enhanced immunoreactivity of the prion protein-specific monoclonal antibody 2A11

Cited by 32 publications

References 31 publications

Sheep-Passaged Bovine Spongiform Encephalopathy Agent Exhibits Altered Pathobiological Properties in Bovine-PrP Transgenic Mice

Sheep-Passaged Bovine Spongiform Encephalopathy Agent Exhibits Altered Pathobiological Properties in Bovine-PrP Transgenic Mice

PrP ^C Governs Susceptibility to Prion Strains in Bank Vole, While Other Host Factors Modulate Strain Features

Subclinical Bovine Spongiform Encephalopathy Infection in Transgenic Mice Expressing Porcine Prion Protein

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Proteinase K enhanced immunoreactivity of the prion protein-specific monoclonal antibody 2A11

Cited by 32 publications

References 31 publications

Sheep-Passaged Bovine Spongiform Encephalopathy Agent Exhibits Altered Pathobiological Properties in Bovine-PrP Transgenic Mice

Sheep-Passaged Bovine Spongiform Encephalopathy Agent Exhibits Altered Pathobiological Properties in Bovine-PrP Transgenic Mice

PrP C Governs Susceptibility to Prion Strains in Bank Vole, While Other Host Factors Modulate Strain Features

Subclinical Bovine Spongiform Encephalopathy Infection in Transgenic Mice Expressing Porcine Prion Protein

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PrP ^C Governs Susceptibility to Prion Strains in Bank Vole, While Other Host Factors Modulate Strain Features