Prion formation, but not clearance, is supported by protein misfolding cyclic amplification

Shikiya, Ronald A.; Eckland, Thomas; Young, Alan J.; Bartz, Jason C.

doi:10.4161/19336896.2014.983759

Cited by 9 publications

(8 citation statements)

References 68 publications

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“…PMCA was also demonstrated as more sensitive than bioassays by several logs of magnitude for the detection of prions [ 36 , 46 , 49 ]. This extended limit of detection might originate from an absence of prion clearance mechanisms in PMCA reactions [ 50 ]. Moreover, PMCA-generated prions appear to be highly infectious, with titers similar, if not identical, to those derived from laboratory bioassays at the terminal stage of the disease [ 36 , 51 ].…”

Section: Discussionmentioning

confidence: 99%

Rapid and Highly Sensitive Detection of Variant Creutzfeldt - Jakob Disease Abnormal Prion Protein on Steel Surfaces by Protein Misfolding Cyclic Amplification: Application to Prion Decontamination Studies

et al. 2016

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The prevalence of variant Creutzfeldt-Jakob disease (vCJD) in the population remains uncertain, although it has been estimated that 1 in 2000 people in the United Kingdom are positive for abnormal prion protein (PrPTSE) by a recent survey of archived appendix tissues. The prominent lymphotropism of vCJD prions raises the possibility that some surgical procedures may be at risk of iatrogenic vCJD transmission in healthcare facilities. It is therefore vital that decontamination procedures applied to medical devices before their reprocessing are thoroughly validated. A current limitation is the lack of a rapid model permissive to human prions. Here, we developed a prion detection assay based on protein misfolding cyclic amplification (PMCA) technology combined with stainless-steel wire surfaces as carriers of prions (Surf-PMCA). This assay allowed the specific detection of minute quantities (10−8 brain dilution) of either human vCJD or ovine scrapie PrPTSE adsorbed onto a single steel wire, within a two week timeframe. Using Surf-PMCA we evaluated the performance of several reference and commercially available prion-specific decontamination procedures. Surprisingly, we found the efficiency of several marketed reagents to remove human vCJD PrPTSE was lower than expected. Overall, our results demonstrate that Surf-PMCA can be used as a rapid and ultrasensitive assay for the detection of human vCJD PrPTSE adsorbed onto a metallic surface, therefore facilitating the development and validation of decontamination procedures against human prions.

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Section: Discussionmentioning

confidence: 99%

Rapid and Highly Sensitive Detection of Variant Creutzfeldt - Jakob Disease Abnormal Prion Protein on Steel Surfaces by Protein Misfolding Cyclic Amplification: Application to Prion Decontamination Studies

et al. 2016

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“…Western blot detection of PrP Sc from brain homogenate was performed as described previously (53). Briefly, brain homogenate (5%, wt/vol) was digested with proteinase K (PK) at a final concentration of 2 U/ml (Roche Diagnostics Corporation, Indianapolis, IN) at 37°C for 60 min.…”

Section: Methodsmentioning

confidence: 99%

“…Hamster prion protein was detected using the mouse monoclonal anti-PrP antibody 3F4 (1:10,000; Chemicon, Temecula, CA). The Western blot was developed with Pierce SuperSignal West Femto maximum-sensitivity substrate, according to the manufacturer's instructions (Pierce, Rockford, IL), and imaged on a Kodak 4000R Imaging Station (Kodak, Rochester, NY) as previously described (53). PrP Sc conformational stability assay.…”

Section: Methodsmentioning

confidence: 99%

Incongruity between Prion Conversion and Incubation Period following Coinfection

Langenfeld

Shikiya

Kincaid

et al. 2016

J Virol

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When multiple prion strains are inoculated into the same host, they can interfere with each other. Strains with long incubation periods can suppress conversion of strains with short incubation periods; however, nothing is known about the conversion of the long-incubation-period strain during strain interference. To investigate this, we inoculated hamsters in the sciatic nerve with long-incubation-period strain 139H prior to superinfection with the short-incubation-period hyper (HY) strain of transmissible mink encephalopathy (TME). First, we found that 139H is transported along the same neuroanatomical tracks as HY TME, adding to the growing body of evidence indicating that PrP Sc favors retrograde transneuronal transport. In contrast to a previous report, we found that 139H interferes with HY TME infection, which is likely due to both strains targeting the same population of neurons following sciatic nerve inoculation. Under conditions where 139H blocked HY TME from causing disease, the strainspecific properties of PrP Sc corresponded with the strain that caused disease, consistent with our previous findings. In the groups of animals where incubation periods were not altered, we found that the animals contained a mixture of 139H and HY TME PrP Sc . This finding expands the definition of strain interference to include conditions where PrP Sc formation is altered yet disease outcome is unaltered. Overall, these results contradict the premise that prion strains are static entities and instead suggest that strain mixtures are dynamic regardless of incubation period or clinical outcome of disease. IMPORTANCEPrions can exist as a mixture of strains in naturally infected animals, where they are able to interfere with the conversion of each other and to extend incubation periods. Little is known, however, about the dynamics of strain conversion under conditions where incubation periods are not affected. We found that inoculation of the same animal with two strains can result in the alteration of conversion of both strains under conditions where the resulting disease was consistent with infection with only a single strain. These data challenge the idea that prion strains are static and suggests that strain mixtures are more dynamic than previously appreciated. This observation has significant implications for prion adaptation. P rion diseases are transmissible neurodegenerative diseases of animals, including humans, with no known effective treatment. Prion diseases of animals include scrapie of sheep and goats, chronic wasting disease of cervids, bovine spongiform encephalopathy of cattle, and transmissible mink encephalopathy (TME) of ranch-raised mink. Prion diseases of humans include kuru of the Fore people of Papua New Guinea, Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome, and fatal familial insomnia (1-8). Human prion diseases are unique in biology in that they can have infectious, familial, or sporadic etiologies. Interestingly, brain material from all three etiologies is infecti...

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“…that balance between formation and clearance) occur in vivo [ 28 , 66 ]. This has been further investigated with PMCA that supports PrP Sc formation but not clearance [ 67 ]. Using PMCA with brain homogenate as the template for conversion, prions strains with short incubation periods generally have more efficient PrP Sc formation compared to PrP Sc from long incubation period strains [ 41 , 68 ].…”

Section: Discussionmentioning

confidence: 99%

PrPSc formation and clearance as determinants of prion tropism

et al. 2017

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Prion strains are characterized by strain-specific differences in neuropathology but can also differ in incubation period, clinical disease, host-range and tissue tropism. The hyper (HY) and drowsy (DY) strains of hamster-adapted transmissible mink encephalopathy (TME) differ in tissue tropism and susceptibility to infection by extraneural routes of infection. Notably, DY TME is not detected in the secondary lymphoreticular system (LRS) tissues of infected hosts regardless of the route of inoculation. We found that similar to the lymphotropic strain HY TME, DY TME crosses mucosal epithelia, enters draining lymphatic vessels in underlying laminae propriae, and is transported to LRS tissues. Since DY TME causes disease once it enters the peripheral nervous system, the restriction in DY TME pathogenesis is due to its inability to establish infection in LRS tissues, not a failure of transport. To determine if LRS tissues can support DY TME formation, we performed protein misfolding cyclic amplification using DY PrPSc as the seed and spleen homogenate as the source of PrPC. We found that the spleen environment can support DY PrPSc formation, although at lower rates compared to lymphotropic strains, suggesting that the failure of DY TME to establish infection in the spleen is not due to the absence of a strain-specific conversion cofactor. Finally, we provide evidence that DY PrPSc is more susceptible to degradation when compared to PrPSc from other lymphotrophic strains. We hypothesize that the relative rates of PrPSc formation and clearance can influence prion tropism.

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Prion formation, but not clearance, is supported by protein misfolding cyclic amplification

Cited by 9 publications

References 68 publications

Rapid and Highly Sensitive Detection of Variant Creutzfeldt - Jakob Disease Abnormal Prion Protein on Steel Surfaces by Protein Misfolding Cyclic Amplification: Application to Prion Decontamination Studies

Rapid and Highly Sensitive Detection of Variant Creutzfeldt - Jakob Disease Abnormal Prion Protein on Steel Surfaces by Protein Misfolding Cyclic Amplification: Application to Prion Decontamination Studies

Incongruity between Prion Conversion and Incubation Period following Coinfection

PrPSc formation and clearance as determinants of prion tropism

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