Dissertation
TRANSCRIPTOMIC DETERMINANTS OF PRION PERMISSIVENESS IN A PATHOPHYSIOLOGICALLY RELEVANT EX VIVO MODEL
Doctor of Philosophy (PhD), Washington State University
01/2015
Handle:
https://hdl.handle.net/2376/112341
Abstract
Susceptibility to infection by prions is highly dependent on the amino acid sequence and host cellular expression of the cellular prion protein (PrPC); however, cellular expression of a genetically susceptible PrPC is insufficient. Thus, additional cellular factors must influence susceptibility to prion infection. The aim of this study was to elucidate the factors associated with permissiveness and resistance to natural scrapie prions in natural TSE host cultured cells. This was accomplished by comparing the global transcriptional profiles of prion-permissive and prion-resistant clones of immortalized ovine microglia using RNA-Seq. First, ovine brain macrophages (microglia) were immortalized by transfection with the human telomerase reverse transcriptase (hTERT) gene to identify cell lines (hTERT-microglia) permissive to natural scrapie prion isolates. Multiple sublines were permissive to cell culture-adapted prions; two sublines were also permissive to natural scrapie isolates (i.e., derived from brain homogenates of sheep infected with scrapie). Then, two ovine microglia clones with different prion permissiveness, but no detectable differences in PrPC expression levels, were inoculated with either scrapie-positive or scrapie-negative sheep brainstem homogenates. Five passages post-inoculation, the transcriptional profiles of mock and infected microglia clones were sequenced using Illumina technology. Comparative transcriptional analyses identified twenty-two differentially transcribed genes, most of which were upregulated in prion-resistant microglia. This included genes encoding for selenoprotein P, endolysosomal proteases, and proteins involved in extracellular matrix remodeling (P < 0.05). Furthermore, in prion permissive microglia, transforming growth factor β-induced, retinoic acid receptor response 1, and phosphoserine aminotranspherase 1 gene transcripts were upregulated (P < 0.05). Our study identifies new genes potentially involved in scrapie prion resistance and susceptibility in a unique natural TSE host ex vivo model, and corroborates results from other studies.
Metrics
5 File views/ downloads
16 Record Views
Details
- Title
- TRANSCRIPTOMIC DETERMINANTS OF PRION PERMISSIVENESS IN A PATHOPHYSIOLOGICALLY RELEVANT EX VIVO MODEL
- Creators
- Juan F. Munoz Gutierrez
- Contributors
- James B Stanton (Advisor)Timothy V Baszler (Committee Member)David A Schneider (Committee Member)Anthony Nicola (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Veterinary Medicine, College of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Number of pages
- 85
- Identifiers
- 99900581730501842
- Language
- English
- Resource Type
- Dissertation