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Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent

Authors
  • Bélondrade, Maxime1
  • Nicot, Simon1
  • Mayran, Charly1
  • Bruyere-Ostells, Lilian1
  • Almela, Florian1
  • Di Bari, Michele A.2
  • Levavasseur, Etienne3
  • Watts, Joel C.4
  • Fournier-Wirth, Chantal1
  • Lehmann, Sylvain5
  • Haïk, Stéphane3
  • Nonno, Romolo2
  • Bougard, Daisy1
  • 1 Université de Montpellier,
  • 2 Istituto Superiore di Sanita,
  • 3 Sorbonne Universités,
  • 4 University of Toronto,
  • 5 Univ Montpellier,
Type
Published Article
Journal
Scientific Reports
Publisher
Springer Nature
Publication Date
Feb 18, 2021
Volume
11
Identifiers
DOI: 10.1038/s41598-021-83630-1
PMID: 33603091
PMCID: PMC7893054
Source
PubMed Central
Keywords
Disciplines
  • Article
License
Unknown

Abstract

Unlike variant Creutzfeldt–Jakob disease prions, sporadic Creutzfeldt–Jakob disease prions have been shown to be difficult to amplify in vitro by protein misfolding cyclic amplification (PMCA). We assessed PMCA of pathological prion protein (PrPTSE) from 14 human sCJD brain samples in 3 substrates: 2 from transgenic mice expressing human prion protein (PrP) with either methionine (M) or valine (V) at position 129, and 1 from bank voles. Brain extracts representing the 5 major clinicopathological sCJD subtypes (MM1/MV1, MM2, MV2, VV1, and VV2) all triggered seeded PrPTSE amplification during serial PMCA with strong seed- and substrate-dependence. Remarkably, bank vole PrP substrate allowed the propagation of all sCJD subtypes with preservation of the initial molecular PrPTSE type. In contrast, PMCA in human PrP substrates was accompanied by a PrPTSE molecular shift during heterologous (M/V129) PMCA reactions, with increased permissiveness of V129 PrP substrate to in vitro sCJD prion amplification compared to M129 PrP substrate. Combining PMCA amplification sensitivities with PrPTSE electrophoretic profiles obtained in the different substrates confirmed the classification of 4 distinct major sCJD prion strains (M1, M2, V1, and V2). Finally, the level of sensitivity required to detect VV2 sCJD prions in cerebrospinal fluid was achieved.

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