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A cell culture model for the propagation of scrapie prion infectivity

Kanu, Nnennaya; (2002) A cell culture model for the propagation of scrapie prion infectivity. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

The central event in the pathogenesis of prion disease is the conversion of PrP, the prion protein, from its normal cellular form (PrPC) to its abnormal conformer (PrPSc). In an experimental scrapie prion infection, the initial inoculum of PrPSc is cleared relatively rapidly, thus a critical aspect of disease progression in vivo is presumably the ability of infected cells, for example in the nervous system, to convert their uninfected neighbours to stable expression of PrPSc. The mechanisms underlying such cell to cell infection are not understood. In the present study, the transfer of scrapie prion infectivity was studied in dissociated cell culture. Genetically marked target cell lines were derived by transfection of tagged PrP molecules into uninfected cells under neomycin selection. In initial experiments, target cells were exposed to subcellular preparations of prion rods and were converted to stable production of PrPSc. In order to study cell-mediated infection, a system was established whereby scrapie-infected mouse SMB cells are able to convert uninfected target cells following coculture of the two populations. The mechanism of intercellular prion transmission in this culture system was evaluated in a series of experiments, and was found to be dependent on direct cell to cell contact. This system for establishing cell-based infection is effective and requires significantly less PrPSc than conversion by an infected brain homogenate. To facilitate the accurate quantitation of conversion at a cellular level, attempts were made to identify a cytochemical marker for PrPSc-expressing cells. This search was pursued by assessing the reactivity of a range of PrP antibodies and reagents including plasminogen. Data obtained from such studies should enable the derivation of immunoreagents able to reliably discriminate between PrPC and PrPSc at the single cell level.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: A cell culture model for the propagation of scrapie prion infectivity
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest.
Keywords: Biological sciences; Health and environmental sciences; Prion diseases
URI: https://discovery.ucl.ac.uk/id/eprint/10104750
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