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Transfer of scrapie prion infectivity by cell contact in culture

Kanu, N; Imokawa, Y; Drechsel, DN; Williamson, RA; Birkett, CR; Bostock, CJ; Brockes, JP; (2002) Transfer of scrapie prion infectivity by cell contact in culture. CURR BIOL , 12 (7) 523 - 530.

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Abstract

Background: When a cell is infected with scrapie prions, newly synthesized molecules of the prion protein PrPC are expressed at the cell surface and may subsequently be converted to the abnormal form PrPSc. In an experimental scrapie infection of an animal, the initial innoculum of PrPsc is cleared relatively rapidly, and the subsequent propagation of the infection depends on the ability of infected cells to convert uninfected target cells to stable production of PrPSc. The mechanism of such cell-based infection is not understood.Results: We have established a system in dissociated cell culture in which scrapie-infected mouse SMB cells are able to stably convert genetically marked target cells by coculture. After coculture and rigorous removal of SMB cells, the target cells express PrPSc and also incorporate [S-35]methionine into PrPSc. The extent of conversion was sensitive to the ratio of the two cell types, and conversion by live SMB required 2500-fold less PrPSc than conversion by a cell-free prion preparation. The conversion activity of SMB cells is not detectable in conditioned medium and apparently depends on close proximity or contact, as evidenced by culturing the SMB and target cells on neighboring but separate surfaces. SMB cells were killed by fixation in aldehydes, followed by washing, and were found to retain significant activity at conversion of target cells.Conclusions: Cell-mediated infection of target cells in this culture system is effective and requires significantly less PrPSc than infection by a prion preparation. Several lines of evidence indicate that it depends on cell contact, in particular, the activity of aldehyde-fixed infected cells.

Type: Article
Title: Transfer of scrapie prion infectivity by cell contact in culture
Keywords: NEUROBLASTOMA-CELLS, MONOCLONAL-ANTIBODY, PROTEIN, EXPRESSION, AGENT, MULTIPLICATION, NEUROINVASION, PATHOGENESIS, LIPOSOMES, STRAINS
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Oncology
URI: http://discovery.ucl.ac.uk/id/eprint/145371
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