l -Arginine ethylester enhances in vitro amplification of PrP Sc in macaques with atypical L-type bovine spongiform encephalopathy and enables presymptomatic detection of PrP Sc in the bodily fluids

Murayama, Yuichi; Ono, Fumiko; Shimozaki, Noriko; Shibata, Hiroaki

doi:10.1016/j.bbrc.2016.01.105

Cited by 7 publications

(9 citation statements)

References 34 publications

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“…Another modified PMCA protocol using L-Arginine ethylester also allowed early detection of prions in saliva as well as in urine of atypical BSE-infected macaques. PrP Sc presence was shown in saliva in half of the samples obtained from one animal at pre-clinical stage of the disease and in all the samples of another animal at the symptomatic phase of the disease [85]. These studies clearly indicate the importance of optimizing each assay for specific prion strains and PrP Sc seed source.…”

Section: Salivamentioning

confidence: 77%

“…PMCA has been applied to the detection of prions in CSF of hamsters infected with scrapie-derived prions, reaching detection limits of approximately 50 attograms PrP Sc [31]. Similarly, but increasing the sensitivity by the use of specific cofactors during the propagation in vitro, CSF from macaques infected with classical BSE and atypical L-type BSE (L-BSE) at the pre-symptomatic stage was also used [84,85]. During the last decade, PMCA has been progressively substituted by the RT-QuIC, for the detection of PrP Sc in CSF of animals affected by prion diseases.…”

Section: Prp Sc In Csf In Animal Prion Diseasesmentioning

confidence: 99%

“…Variations of the PMCA methodology in order to enhance sensitivity have been used for the specific detection of atypical BSE prions in urine of experimentally infected macaques. Using L-Arginine ethylester during the in vitro propagation, the technique allowed detection of prions in urine of 1 out of 2 animals at pre-clinical stage and in all the samples at the clinical stage of disease [85]. PMCA has also been applied to the study of scrapie-affected sheep and CWD-affected deer urine samples.…”

Section: Urinementioning

confidence: 99%

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Detection of Pathognomonic Biomarker PrPSc and the Contribution of Cell Free-Amplification Techniques to the Diagnosis of Prion Diseases

et al. 2020

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Transmissible spongiform encephalopathies or prion diseases are rapidly progressive neurodegenerative diseases, the clinical manifestation of which can resemble other promptly evolving neurological maladies. Therefore, the unequivocal ante-mortem diagnosis is highly challenging and was only possible by histopathological and immunohistochemical analysis of the brain at necropsy. Although surrogate biomarkers of neurological damage have become invaluable to complement clinical data and provide more accurate diagnostics at early stages, other neurodegenerative diseases show similar alterations hindering the differential diagnosis. To solve that, the detection of the pathognomonic biomarker of disease, PrP Sc , the aberrantly folded isoform of the prion protein, could be used. However, the amounts in easily accessible tissues or body fluids at pre-clinical or early clinical stages are extremely low for the standard detection methods. The solution comes from the recent development of in vitro prion propagation techniques, such as Protein Misfolding Cyclic Amplification (PMCA) and Real Time-Quaking Induced Conversion (RT-QuIC), which have been already applied to detect minute amounts of PrP Sc in different matrixes and make early diagnosis of prion diseases feasible in a near future. Herein, the most relevant tissues and body fluids in which PrP Sc has been detected in animals and humans are being reviewed, especially those in which cell-free prion propagation systems have been used with diagnostic purposes.

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Section: Salivamentioning

confidence: 77%

Section: Prp Sc In Csf In Animal Prion Diseasesmentioning

confidence: 99%

Section: Urinementioning

confidence: 99%

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Detection of Pathognomonic Biomarker PrPSc and the Contribution of Cell Free-Amplification Techniques to the Diagnosis of Prion Diseases

et al. 2020

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“…To this end, several studies have looked to develop in vitro prion amplification assays to detect very low levels of atypical BSE prions. Murayama and coworkers used wild type mouse brain as sPMCA substrate supplemented with a cocktail of polyanions (polyadenylic acid salt (Poly-A), heparin and sodium polyphosphate) and L-arginine ethylester 16 . The method included sPMCA amplifications over 280 hours and could detect L-BSE down to a 10 −10 dilution of brain homogenate.…”

Section: Discussionmentioning

confidence: 99%

“…To date, PMCA has been used for the high sensitivity detection of L-BSE derived from experimentally infected macaques although the products were not distinguished from other types of BSE 16 . QuIC has also been used to amplify both L-BSE and H-BSE from infected cattle.…”

Section: Introductionmentioning

confidence: 99%

In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification

O’Connor

Bishop

Workman

et al. 2017

Prion

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The in vitro amplification of prions by serial protein misfolding cyclic amplification has been shown to detect PrPSc to levels at least as sensitive as rodent bioassay but in a fraction of the time. Bovine spongiform encephalopathy is a zoonotic prion disease in cattle and has been shown to occur in 3 distinct forms, classical BSE (C-BSE) and 2 atypical BSE forms (L-BSE and H-BSE). Atypical forms are usually detected in asymptomatic, older cattle and are suggested to be spontaneous forms of the disease. Here, we show the development of a serial protein misfolding cyclic amplification method for the detection of H-BSE. The assay could detect PrPSc from 3 distinct experimental isolates of H-BSE, could detect PrPSc in as little as 1×10−12 g of brain material and was highly specific. Additionally, the product of serial protein misfolding cyclic amplification at all dilutions of seed analyzed could be readily distinguished from L-BSE, which did not amplify, and C-BSE, which had PrPSc with distinct protease K-resistance and protease K-resistant PrPSc molecular weights.

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Heparan Sulfate and Heparin Promote Faithful Prion Replication in Vitro by Binding to Normal and Abnormal Prion Proteins in Protein Misfolding Cyclic Amplification

Imamura

Tabeta

Kato

et al. 2016

Journal of Biological Chemistry

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Edited by Paul FraserThe precise mechanism underlying the conversion of normal prion protein ( Sc by using a modified PMCA performed with baculovirusderived recombinant PrP (Bac-PrP) as a substrate. The addition of heparan sulfate (HS) or its analog heparin (HP) restored the conversion efficiency in PMCA that was inhibited through nucleic acid depletion. Moreover, the PMCA products obtained under these conditions were infectious and preserved the properties of the input PrP Sc . These data suggest that HS and HP play the same role as nucleic acids in facilitating faithful replication of prions in PMCA. Furthermore, we showed that HP binds to both Bac-PrP and Bac-PrP Sc through the sulfated groups present on HP and that the N-terminal domain of Bac-PrP Sc might potentially not be involved in the binding to HP. These results suggest that the interaction of GAGs such as HS and HP with PrP C and/or PrP Sc through their sulfate groups is critical for the faithful replication of prions.

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l -Arginine ethylester enhances in vitro amplification of PrP Sc in macaques with atypical L-type bovine spongiform encephalopathy and enables presymptomatic detection of PrP Sc in the bodily fluids

Cited by 7 publications

References 34 publications

Detection of Pathognomonic Biomarker PrPSc and the Contribution of Cell Free-Amplification Techniques to the Diagnosis of Prion Diseases

Detection of Pathognomonic Biomarker PrPSc and the Contribution of Cell Free-Amplification Techniques to the Diagnosis of Prion Diseases

In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification

Heparan Sulfate and Heparin Promote Faithful Prion Replication in Vitro by Binding to Normal and Abnormal Prion Proteins in Protein Misfolding Cyclic Amplification

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