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Volume 24, Number 8—August 2018
Research

Susceptibility of Human Prion Protein to Conversion by Chronic Wasting Disease Prions

Marcelo A. BarriaComments to Author , Adriana Libori, Gordon Mitchell, and Mark W. Head
Author affiliations: National CJD Research and Surveillance Unit, University of Edinburgh, Edinburgh, Scotland, UK (M.A. Barria, A. Libori, M.W. Head); National and OIE Reference Laboratory for Scrapie and CWD, Canadian Food Inspection Agency, Ottawa, Ontario, Canada (G. Mitchell)

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Figure 4

Evaluation of the in vitro conversion of human prion protein (PrP) seeded with the misfolded, disease-associated prion protein form present in chronic wasting disease (CWD)–affected reindeer brain samples. We incubated 2 reindeer CWD specimens (reindeer 1 and 2) in a panel of 3 humanized transgenic substrates (Tg-HuMM, Tg-HuMV, and Tg-HuVV) and subjected them to a single round of protein misfolding cyclic amplification (PMCA). We diluted PMCA seeds 3 times in fresh PMCA substrate (dilution facto

Figure 4. Evaluation of the in vitro conversion of human prion protein (PrP) seeded with the misfolded, disease-associated prion protein form present in chronic wasting disease (CWD)–affected reindeer brain samples. We incubated 2 reindeer CWD specimens (reindeer 1 and 2) in a panel of 3 humanized transgenic substrates (Tg-HuMM, Tg-HuMV, and Tg-HuVV) and subjected them to a single round of protein misfolding cyclic amplification (PMCA). We diluted PMCA seeds 3 times in fresh PMCA substrate (dilution factor 1:3) and evaluated PMCA reactions for the presence of protease-resistant prion protein (PrPres) by proteinase K digestion. We performed Western blot analysis by using mAb 3F4 (for the detection of human PrPres) and mAb 6H4 (for detection of CWD PrPres and human PrPres). We incorporated the elk specimen designated elk 0 as a control. We performed >5 repeats for the amplification of reindeer 1 and 2 specimens. Reference molecular markers have been included. Molecular mass of electrophoretic markers is given. Odd and even number lanes show reaction mixtures before and after PMCA. mAb, monoclonal antibody; Tg-HuMM, humanized transgenic PRNP codon 129 homozygous methionine; Tg-Hu-MV, humanized transgenic methionine/valine; Tg-HuVV, humanized transgenic valine/valine.

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Page created: July 17, 2018
Page updated: July 17, 2018
Page reviewed: July 17, 2018
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