A short purification process for quantitative isolation of PrPScfrom naturally occurring and experimental transmissible spongiform encephalopathies

Polymenidou, Magdalini; Verghese-Nikolakaki, S.; Groschup, Martin H.; Chaplin, Melanie J.; Stack, M.J.; Plaitakis, Andreas; Sklaviadis, Theodoros

doi:10.1186/1471-2334-2-23

Cited by 23 publications

(19 citation statements)

References 32 publications

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“…Solubilized recombinant PrP as well as unprocessed or enriched in PrP Sc [32] brain homogenates from terminally ill mice were analyzed by western blotting. For western blotting, proteins were resolved onto 13% polyacrylamide gels and then electrotransferred onto polyvinylidenefluoride membranes (PVDF, Millipore).…”

Section: Methodsmentioning

confidence: 99%

Immunization with Recombinant Prion Protein Leads to Partial Protection in a Murine Model of TSEs through a Novel Mechanism

et al. 2013

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Transmissible spongiform encephalopathies are neurodegenerative diseases, which despite fervent research remain incurable. Immunization approaches have shown great potential at providing protection, however tolerance effects hamper active immunization protocols. In this study we evaluated the antigenic potential of various forms of recombinant murine prion protein and estimated their protective efficacy in a mouse model of prion diseases. One of the forms tested provided a significant elongation of survival interval. The elongation was mediated via an acute depletion of mature follicular dendritic cells, which are associated with propagation of the prion infectious agent in the periphery and in part to the development of humoral immunity against prion protein. This unprecedented result could offer new strategies for protection against transmissible encephalopathies as well as other diseases associated with follicular dendritic cells.

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Section: Methodsmentioning

confidence: 99%

Immunization with Recombinant Prion Protein Leads to Partial Protection in a Murine Model of TSEs through a Novel Mechanism

et al. 2013

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“…All animals were killed at the terminal stages of the disease by brief anaesthesia using ether and then cervical dislocation. The presence of PrP Sc in brain tissue from both β4 –/– and wild‐type C57/Bl6 mice was verified using a short purification protocol, as previously described (Polymenidou et al ., 2002). Survival curves were drawn using GraphPad Prism software.…”

Section: Methodsmentioning

confidence: 99%

Cellular prion protein co‐localizes with nAChR β4 subunit in brain and gastrointestinal tract

Petrakis¹,

Irinopoulou²,

Panagiotidis³

et al. 2008

Eur J of Neuroscience

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PrP(C), the cellular isoform of prion protein, is widely expressed in most tissues, including brain, muscle and gastrointestinal tract. Despite its involvement in several bioprocesses, PrP has still no apparent physiological role. During propagation of transmissible spongiform encephalopathies (TSE), prion protein is converted to the pathological isoform, PrP(Sc), in a process believed to be mediated by unknown host factors. The identification of proteins associated with PrP may provide information about both the biology of prions and the pathogenesis of TSE. Thus far, PrP(C) has been shown to interact with synaptic proteins, components of the cytoskeleton and intracellular proteins involved in signalling pathways. Here, we describe the association of PrP with the beta4 subunit of nicotinic acetylcholine receptor (nAChR), as indicated by co-immunoprecipitation assays and double-label immunofluorescence. The interaction between prion protein and native beta4 subunit was further studied by affinity chromatography, using immobilized and refolded recombinant PrP as a bait and brain homogenates from normal individuals. Additionally, the participation of beta4 subunit in the pathogenesis of TSE was studied by in vivo assays. beta4(-/-) and wild-type mice were challenged with the RML (Rocky Mountain Laboratories) infectious agent. Transgenic animals displayed altered incubation times but the deletion of beta4 subunit did not result in a significant change of the incubation period of the disease. Our results suggest that PrP(C) is a member of a multiprotein membrane complex participating in the formation and function of alpha3beta4 nAChR.

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“…Nickel powder may therefore find a use in detection and/or removal of prions in large volumes of fluids such as urine, abattoir wastewater or other sources where there is a risk of prion contamination. Several methods have been designed for PrP Sc concentration, such as immunoprecipitation, TCA precipitation, salt precipitation (Hilmert & Diringer, 1984;Polymenidou et al, 2002), sodium phosphotungstate or streptomycin precipitation (Moussa et al, 2006). PrP Sc bound to stainless steel retains its infectivity (Flechsig et al, 2001) and therefore, most likely, its original conformation.…”

Section: Discussionmentioning

confidence: 99%

Prion adsorption to stainless steel is promoted by nickel and molybdenum

Luhr

Lőw

Taraboulos

et al. 2009

Journal of General Virology

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Prions are infectious agents resulting from the conversion of a normal cellular protein, PrP C , to a misfolded species, PrPSc . Iatrogenic transmission of prions is known from surgical procedures involving stainless steel materials. Here, it was shown that stainless steel containing nickel and molybdenum binds PrP Sc more efficiently and transmits infection to cells in culture to a higher degree than if these elements are not present. Furthermore, both nickel and molybdenum alone adsorbed PrP Sc , and nickel powder could be used to extract PrP Sc from dilute solutions, thus providing a simple approach to concentration of PrP Sc . The fact that nickel and molybdenum in steel alloys increased the binding affinity, and bound infectivity, of PrP Sc is an important issue to consider in the manufacture of surgical instruments and abattoir tools. INTRODUCTIONPrions cause lethal neurodegenerative diseases such as scrapie in sheep, bovine spongiform encephalopathy in cows and Creutzfeldt-Jakob disease (CJD) in humans. The infectious agents are essentially composed of abnormally folded, partially protease-resistant proteins, PrP Sc , rich in b-sheet structure, that can impose their conformation on the normal cellular prion proteins, PrP C (Prusiner, 1998). PrP C and PrP Sc can both exist as full-length proteins, but N-terminal trimming of PrP Sc by limited proteolysis in vitro or by cellular proteases generates a 27-30 kDa fragment, PrP 27-30, which is a proteinase K (PK)-resistant product that retains infectivity (Gabizon et al., 1988;Meyer et al., 1986).Infectious prions are present at high levels in the brain, spinal cord, retina, tonsils and spleen (Bruce et al., 2001;Hogan et al., 1986; Wadsworth et al., 2001), but also at lower levels in blood and skeletal muscles (Mulcahy et al., 2004;Peden et al., 2004Peden et al., , 2006. Prions are resistant to conventional sterilization procedures and to inactivation by various chemical treatments (Alper et al., 1966;Gibbs et al., 1994Gibbs et al., , 1978. The potential presence of prions in biological tissues and fluids has led to the awareness that instruments and materials used in dentistry, surgery and abattoir activities may be a source of prion spread. For instance, prion-contaminated intracerebrally implanted electrodes can transmit CJD in humans (Bernoulli et al., 1977;Gibbs et al., 1994), and stainless steel suture wires exposed to scrapie can efficiently transmit the infection to indicator mice (Flechsig et al., 2001;Zobeley et al., 1999) and in this context can be used as transfer vehicles in bioassays for prion infectivity (Fichet et al., 2004;Lemmer et al., 2004;Yan et al., 2004). To minimize the risk of prion transmission, it is important to devise novel methods of disinfecting stainless steel instruments and materials (Baier et al., 2004; Fichet et al., 2004;Jackson et al., 2005; MullerHellwig et al., 2006;Peretz et al., 2006;Race & Raymond, 2004;Yan et al., 2004), as many existing decontamination methods may damage the instrumentation (Brown et al., 2005). Howe...

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A short purification process for quantitative isolation of PrPScfrom naturally occurring and experimental transmissible spongiform encephalopathies

Cited by 23 publications

References 32 publications

Immunization with Recombinant Prion Protein Leads to Partial Protection in a Murine Model of TSEs through a Novel Mechanism

Immunization with Recombinant Prion Protein Leads to Partial Protection in a Murine Model of TSEs through a Novel Mechanism

Cellular prion protein co‐localizes with nAChR β4 subunit in brain and gastrointestinal tract

Prion adsorption to stainless steel is promoted by nickel and molybdenum

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