NR AWGL
AU Harris,R.; Booth,S.A.
TI Differential gene expression studies in murine splenic follicular dendritic cells during prion infection
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PA-20
PT Konferenz-Poster
AB During pre-clinical infection PrPsc has been shown to accumulate in the secondary lymphoid tissues of infected humans and animals. PrPsc builds up in germinal centres and is strongly associated with follicular dendritic cells (FDCs). Additionally, studies have shown that mature FDCs are critical for PrPsc replication in lymphoid tissues, and dedifferentiation of the cells reduces scrapie susceptibility. Analysis of gene expression differences in lymphoid tissues may allow us to gain valuable insight into prion biology and the mechanisms of neuroinvasion. We have identified over 100 genes that are differentially expressed in the spleen during preclinical stages of the disease. We will pinpoint the cellular location of these gene expression changes in the spleen, in particular focusing on gene expression in FDCs during replication of infectious prion protein. Our objective is to use DNA microarrays to obtain gene expression profiles from FDCs, with the aim of identifying host factors involved in prion replication and also identifying preclinical biomarkers for diagnosis. Accordingly, mice were infected with scrapie and at the endpoint of infection the spleens were extracted. The spleens were then homogenized and a population of splenic FDCs were enriched. Total RNA was prepared from infected and non-infected cells and amplified using Eberwine linear amplification methods to generate enough target material for dual-color, competitive hybridization to a DNA microarray. The microarray platforms used in this project are custom made in our laboratory and comprised of over 17K genes of libraries made mostly from mouse CNS genes. The microarray data was analyzed using the latest computational statistical software available in order to identify differentially expressed genes during the course of the disease. Ultimately, the expression data will lead to the identification of specific biomarkers. Preliminary results have shown from studying noninfected mice that an enriched population of FDCs can be isolated from the spleen and their expression profiles are different from that of the whole spleen. We will now move on to studying the infected mice FDC population and compare their diseased expression profiles with what we have found with the non-infected FDCs.
AD Faculty of Medicine, Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, MB, Canada, R3E 0W3; and Public Health Agency of Canada, Division of Host Genetics and Prion Diseases, Winnipeg, MB, Canada R3E 3R2; E-mail: Rhiannon_Harris@phac-aspc.gc.ca
SP englisch
PO Italien