NR AXKJ
AU Charveriat,M.; Reboul,M.; Lenuzza,N.; Picoli,C.; Nouvel,V.; Aubry,F.; Correia,E.; Eterpi,M.; Deslys,J.P.; Mouthon,F.
TI Attempts to Develop Cellular Models of Human Prion Infection
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.122
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Cellular responses to prion infection are largely unknown, especially for human cells, which is partially due to the lack of cellular models infected by human Prions. Our goal was to develop cellular models infectable by human strains, using approaches aimed at 1) the development of new infection protocols and methods, 2) the adaptation of Prion strains to cells; and 3) modifications of cells to replicate Prions.
First, we verified on about forty cell lines from different species and tissues, using different infected human brain homogenates, that no PrPres accumulation could be obtained with classical protocols of infection. We then tested several other protocols on MM129 human cells varying methods of homogenate preparation, infectious load and culture conditions. Notably, we tried to infect human neuroblastomas with arrested growth, cultivated alone or with glial lines (human glioblastomas and astrocytomas) to mimic the in vivo situation in the brain.
In the second approach, to mimic the adaptation of strains to the host, we inoculated human cell cultures with different human TSE strains and on passage examined one part of the cell lysate for PrPres and used the remainder to establish the next passage.
As the two previous approaches relied on the assumption that the human cells tested were susceptible to infection (which may not be correct), we have also tested the possibility to modify the susceptibility of the cells to infection. We developed techniques to change the pattern of gene expression and to generate random genetic mutations. The modified cells were then exposed to different strains and PrPres accumulation was evaluated after several passages.
The experiments are ongoing and the preliminary data will be presented.
AD M. Charveriat, M. Reboul, N. Lenuzza, C. Picoli, V. Nouvel, E. Correia, M. Eterpi, J.P. Deslys, F. Mouthon, CEA/DSV/IMETI/SEPIA, France; F. Aubry, Alliance Biosecure Foundation, France
SP englisch
PO Schottland
EA pdf-Datei und Poster (Postertitel: Screening of anti-prion drugs and establishment of cellular models infected by human prions)