2013 Fiscal Year Final Research Report

Characterization of Creutzfeldt-Jakob disease strains with protein misfolfing cyclic amplification

Research Project

Project/Area Number	24790871
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Neurology
Research Institution	Tohoku University
Principal Investigator	TAKEUCHI Atsuko 東北大学, 医学(系)研究科(研究院), 助教 (00535239)
Project Period (FY)	2012-04-01 – 2014-03-31
Keywords	クロイツフェルトヤコブ病 / プリオン / PMCA
Research Abstract	Protein Misfolding Cyclic Amplification (PMCA) is currently one of the most sensitive method of detecting PrPSc, by which protease-resistant isoform of prion protein can be amplified in vitro. However, when human brains were used as substrate, the efficiency of sporadic Creutzfeldt-Jakob CJD (sCJD) prions was considerably lower than that of scrapie-derived hamster-adapted strain. Then, we used human cell lysates overexpressing human PrPC as substrate instead of human brains (cell-PMCA), resulting in the improvement of amplification efficacy of some types of CJD prions. In this study, we amplified CJD prions from a large number of CJD cases including various types with cPMCA. As a result, prions from MV2, VV2 and dura-mata grafted CJD (dCJD) showing the plaque type depositions of PrPSc (p-dCJD) in the brains were strongly amplified with 129V substrate. Interestingly, p-dCJD prions could be amplified efficiently with 129V substrates whereas the genotype of these cases was 129M/M.