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.

Details

Title
Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent
Author
Bélondrade Maxime 1 ; Nicot, Simon 1 ; Mayran Charly 1 ; Bruyere-Ostells Lilian 1 ; Almela Florian 1 ; Di Bari Michele A 2 ; Levavasseur Etienne 3 ; Watts, Joel C 4 ; Fournier-Wirth, Chantal 1 ; Lehmann, Sylvain 5 ; Haïk Stéphane 3 ; Nonno Romolo 2 ; Bougard Daisy 1 

 Université de Montpellier, Pathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Montpellier, France (GRID:grid.121334.6) (ISNI:0000 0001 2097 0141) 
 Istituto Superiore di Sanita, Department of Food Safety, Nutrition and Veterinary Public Health, Rome, Italy (GRID:grid.416651.1) (ISNI:0000 0000 9120 6856) 
 Sorbonne Universités, Inserm U 1127, CNRS UMR 7225, UPMC Université Paris 06 UMR S 1127, Institut du Cerveau et de la Moelle épinière, Paris, France (GRID:grid.462844.8) (ISNI:0000 0001 2308 1657) 
 University of Toronto, Tanz Centre for Research in Neurodegenerative Diseases and Department of Biochemistry, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
 Univ Montpellier, IRMB, INM, INSERM, CHU Montpellier, (LBPC-PPC), Montpellier, France (GRID:grid.121334.6) (ISNI:0000 0001 2097 0141) 
Publication year
2021
Publication date
2021
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2490849816
Copyright
© The Author(s) 2021. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.