<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification

ABSTRACT Substantial evidence supports the hypothesis that prions are misfolded, infectious, insoluble, and protease-resistant proteins (PrPRES) devoid of instructional nucleic acid that cause transmissible spongiform encephalopathies (TSEs). Protein misfolding cyclic amplification (PMCA) has provid...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Crystal Meyerett-Reid, A. Christy Wyckoff, Terry Spraker, Bruce Pulford, Heather Bender, Mark D. Zabel
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	chronic wasting disease prions protein misfolding cyclic amplification spontaneous generation strains Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/223966d16ccb44c693a8127d11af7d11
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:223966d16ccb44c693a8127d11af7d11
record_format	dspace
spelling	oai:doaj.org-article:223966d16ccb44c693a8127d11af7d112021-11-15T15:22:03Z<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification10.1128/mSphere.00372-162379-5042https://doaj.org/article/223966d16ccb44c693a8127d11af7d112017-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00372-16https://doaj.org/toc/2379-5042ABSTRACT Substantial evidence supports the hypothesis that prions are misfolded, infectious, insoluble, and protease-resistant proteins (PrPRES) devoid of instructional nucleic acid that cause transmissible spongiform encephalopathies (TSEs). Protein misfolding cyclic amplification (PMCA) has provided additional evidence that PrPRes acts as a template that can convert the normal cellular prion protein (PrPC) present in uninfected normal brain homogenate (NBH) into the infectious misfolded PrPRES isoform. Human PrPC has been shown to spontaneously convert to a misfolded pathological state causing sporadic Creutzfeldt-Jakob disease (sCJD). Several investigators have reported spontaneous generation of prions by in vitro assays, including PMCA. Here we tested the rate of de novo generation of cervid prions in our laboratory using our standard PMCA protocol and NBH from transgenic mice expressing cervid PrPC (TgCerPrP mice). We generated de novo prions in rounds 4, 5, and 7 at low cumulative rates of 1.6, 5.0, and 6.7%, respectively. The prions caused infectious chronic wasting disease (CWD) upon inoculation into normal uninfected TgCerPrP mice and displayed unique biochemical characteristics compared to other cervid prion strains. We conclude that PMCA of cervid PrPC from normal brain homogenate spontaneously generated a new cervid prion strain. These data support the potential for cervids to develop sporadic CWD. IMPORTANCE CWD is the only known TSE that affects free-ranging wildlife, specifically cervids such as elk, deer, moose, caribou, and reindeer. CWD has become endemic in both free-ranging and captive herds in North America, South Korea, and, most recently, northern Europe. The prion research community continues to debate the origins of CWD. Original foci of CWD emergence in Colorado and Wyoming coincident with the sheep TSE scrapie suggest that scrapie prions may have adapted to cervids to cause CWD. However, emerging evidence supports the idea that cervid PrPC may be more prone to misfolding to the pathological isoform. Here we test the hypothesis that cervid PrPC can spontaneously misfold to create de novo prions. Whether CWD can arise spontaneously as a sporadic TSE or represents a new TSE caused by cervid-adapted scrapie prions profoundly impacts surveillance and mitigation strategies. Podcast: A podcast concerning this article is available.Crystal Meyerett-ReidA. Christy WyckoffTerry SprakerBruce PulfordHeather BenderMark D. ZabelAmerican Society for Microbiologyarticlechronic wasting diseaseprionsprotein misfolding cyclic amplificationspontaneous generationstrainsMicrobiologyQR1-502ENmSphere, Vol 2, Iss 1 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	chronic wasting disease prions protein misfolding cyclic amplification spontaneous generation strains Microbiology QR1-502
spellingShingle	chronic wasting disease prions protein misfolding cyclic amplification spontaneous generation strains Microbiology QR1-502 Crystal Meyerett-Reid A. Christy Wyckoff Terry Spraker Bruce Pulford Heather Bender Mark D. Zabel <italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification
description	ABSTRACT Substantial evidence supports the hypothesis that prions are misfolded, infectious, insoluble, and protease-resistant proteins (PrPRES) devoid of instructional nucleic acid that cause transmissible spongiform encephalopathies (TSEs). Protein misfolding cyclic amplification (PMCA) has provided additional evidence that PrPRes acts as a template that can convert the normal cellular prion protein (PrPC) present in uninfected normal brain homogenate (NBH) into the infectious misfolded PrPRES isoform. Human PrPC has been shown to spontaneously convert to a misfolded pathological state causing sporadic Creutzfeldt-Jakob disease (sCJD). Several investigators have reported spontaneous generation of prions by in vitro assays, including PMCA. Here we tested the rate of de novo generation of cervid prions in our laboratory using our standard PMCA protocol and NBH from transgenic mice expressing cervid PrPC (TgCerPrP mice). We generated de novo prions in rounds 4, 5, and 7 at low cumulative rates of 1.6, 5.0, and 6.7%, respectively. The prions caused infectious chronic wasting disease (CWD) upon inoculation into normal uninfected TgCerPrP mice and displayed unique biochemical characteristics compared to other cervid prion strains. We conclude that PMCA of cervid PrPC from normal brain homogenate spontaneously generated a new cervid prion strain. These data support the potential for cervids to develop sporadic CWD. IMPORTANCE CWD is the only known TSE that affects free-ranging wildlife, specifically cervids such as elk, deer, moose, caribou, and reindeer. CWD has become endemic in both free-ranging and captive herds in North America, South Korea, and, most recently, northern Europe. The prion research community continues to debate the origins of CWD. Original foci of CWD emergence in Colorado and Wyoming coincident with the sheep TSE scrapie suggest that scrapie prions may have adapted to cervids to cause CWD. However, emerging evidence supports the idea that cervid PrPC may be more prone to misfolding to the pathological isoform. Here we test the hypothesis that cervid PrPC can spontaneously misfold to create de novo prions. Whether CWD can arise spontaneously as a sporadic TSE or represents a new TSE caused by cervid-adapted scrapie prions profoundly impacts surveillance and mitigation strategies. Podcast: A podcast concerning this article is available.
format	article
author	Crystal Meyerett-Reid A. Christy Wyckoff Terry Spraker Bruce Pulford Heather Bender Mark D. Zabel
author_facet	Crystal Meyerett-Reid A. Christy Wyckoff Terry Spraker Bruce Pulford Heather Bender Mark D. Zabel
author_sort	Crystal Meyerett-Reid
title	<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification
title_short	<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification
title_full	<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification
title_fullStr	<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification
title_full_unstemmed	<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification
title_sort	<italic toggle="yes">de novo</italic> generation of a unique cervid prion strain using protein misfolding cyclic amplification
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/223966d16ccb44c693a8127d11af7d11
work_keys_str_mv	AT crystalmeyerettreid italictoggleyesdenovoitalicgenerationofauniquecervidprionstrainusingproteinmisfoldingcyclicamplification AT achristywyckoff italictoggleyesdenovoitalicgenerationofauniquecervidprionstrainusingproteinmisfoldingcyclicamplification AT terryspraker italictoggleyesdenovoitalicgenerationofauniquecervidprionstrainusingproteinmisfoldingcyclicamplification AT brucepulford italictoggleyesdenovoitalicgenerationofauniquecervidprionstrainusingproteinmisfoldingcyclicamplification AT heatherbender italictoggleyesdenovoitalicgenerationofauniquecervidprionstrainusingproteinmisfoldingcyclicamplification AT markdzabel italictoggleyesdenovoitalicgenerationofauniquecervidprionstrainusingproteinmisfoldingcyclicamplification
_version_	1718428057741557760

<italic toggle="yes">De Novo</italic> Generation of a Unique Cervid Prion Strain Using Protein Misfolding Cyclic Amplification

Ejemplares similares