scholarly journals Bioassay of prion-infected blood plasma in PrP transgenic Drosophila

2016 ◽  
Vol 473 (23) ◽  
pp. 4399-4412 ◽  
Author(s):  
Alana M. Thackray ◽  
Olivier Andreoletti ◽  
Raymond Bujdoso
Keyword(s):  
Brain Homogenate ◽  
Infected Sheep ◽  
Mammalian Host ◽  
Infected Blood ◽  
Clinical Phase ◽  
Prion Infectivity ◽  
Sheep Brain ◽  
Blood Fraction ◽  
Transgenic Drosophila ◽  
Mammalian Prions

In pursuit of a tractable bioassay to assess blood prion infectivity, we have generated prion protein (PrP) transgenic Drosophila, which show a neurotoxic phenotype in adulthood after exposure to exogenous prions at the larval stage. Here, we determined the sensitivity of ovine PrP transgenic Drosophila to ovine prion infectivity by exposure of these flies to a dilution series of scrapie-infected sheep brain homogenate. Ovine PrP transgenic Drosophila showed a significant neurotoxic response to dilutions of 10−2 to 10−10 of the original scrapie-infected sheep brain homogenate. Significantly, we determined that this prion-induced neurotoxic response in ovine PrP transgenic Drosophila was transmissible to ovine PrP transgenic mice, which is indicative of authentic mammalian prion detection by these flies. As a consequence, we considered that PrP transgenic Drosophila were sufficiently sensitive to exogenous mammalian prions to be capable of detecting prion infectivity in the blood of scrapie-infected sheep. To test this hypothesis, we exposed ovine PrP transgenic Drosophila to scrapie-infected plasma, a blood fraction notoriously difficult to assess by conventional prion bioassays. Notably, pre-clinical plasma from scrapie-infected sheep induced neurotoxicity in PrP transgenic Drosophila and this effect was more pronounced after exposure to samples collected at the clinical phase of disease. The neurotoxic phenotype in ovine PrP transgenic Drosophila induced by plasma from scrapie-infected sheep was transmissible since head homogenate from these flies caused neurotoxicity in recipient flies during fly-to-fly transmission. Our data show that PrP transgenic Drosophila can be used successfully to bioassay prion infectivity in blood from a prion-diseased mammalian host.

scholarly journals Removal of the glycosylphosphatidylinositol anchor from PrPSc by cathepsin D does not reduce prion infectivity

2006 ◽  
Vol 395 (2) ◽  
pp. 443-448 ◽  
Author(s):  
Patrick A. Lewis ◽  
Francesca Properzi ◽  
Kanella Prodromidou ◽  
Anthony R. Clarke ◽  
John Collinge ◽  
...  
Keyword(s):  
Cathepsin D ◽  
Rocky Mountain ◽  
Brain Homogenate ◽  
Mouse Bioassay ◽  
Gpi Anchor ◽  
Glycosylphosphatidylinositol Anchor ◽  
Prion Infectivity ◽  
C Terminus ◽  
Prion Propagation

According to the protein-only hypothesis of prion propagation, prions are composed principally of PrPSc, an abnormal conformational isoform of the prion protein, which, like its normal cellular precursor (PrPC), has a GPI (glycosylphosphatidylinositol) anchor at the C-terminus. To date, elucidating the role of this anchor on the infectivity of prion preparations has not been possible because of the resistance of PrPSc to the activity of PI-PLC (phosphoinositide-specific phospholipase C), an enzyme which removes the GPI moiety from PrPC. Removal of the GPI anchor from PrPSc requires denaturation before treatment with PI-PLC, a process that also abolishes infectivity. To circumvent this problem, we have removed the GPI anchor from PrPSc in RML (Rocky Mountain Laboratory)-prion-infected murine brain homogenate using the aspartic endoprotease cathepsin D. This enzyme eliminates a short sequence at the C-terminal end of PrP to which the GPI anchor is attached. We found that this modification has no effect (i) on an in vitro amplification model of PrPSc, (ii) on the prion titre as determined by a highly sensitive N2a-cell based bioassay, or (iii) in a mouse bioassay. These results show that the GPI anchor has little or no role in either the propagation of PrPSc or on prion infectivity.

scholarly journals Detection of apoptosis induced DNA cleavage in scrapie-infected sheep brain

1994 ◽  
Vol 115 (2-3) ◽  
pp. 341-346 ◽  
Author(s):  
Daryl W. Fairbairn ◽  
Kevin G. Carnahan ◽  
Richard N. Thwaits ◽  
Richard V. Thwaits ◽  
G. Reed Holyoak ◽  
...  
Keyword(s):  
Dna Cleavage ◽  
Infected Sheep ◽  
Sheep Brain

scholarly journals Long-Term Incubation PrPCWD with Soils Affects Prion Recovery but Not Infectivity

Pathogens ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 311
Author(s):  
Alsu Kuznetsova ◽  
Debbie McKenzie ◽  
Catherine Cullingham ◽  
Judd M. Aiken
Keyword(s):  
Prion Disease ◽  
Soil Type ◽  
Infectious Agent ◽  
Brain Homogenate ◽  
Wasting Disease ◽  
Soil Minerals ◽  
Prion Infectivity ◽  
Incubation Periods ◽  
Clinical Stages

Chronic wasting disease (CWD) is a contagious prion disease of cervids. The infectious agent is shed from animals at the preclinical and clinical stages of disease where it persists in the environment as a reservoir of CWD infectivity. In this study, we demonstrate that long-term incubation of CWD prions (generated from tg-mice infected with deer or elk prions) with illite, montmorillonite (Mte) and whole soils results in decreased recovery of PrPCWD, suggesting that binding becomes more avid and irreversible with time. This continual decline of immunoblot PrPCWD detection did not correlate with prion infectivity levels. Bioassay showed no significant differences in incubation periods between mice inoculated with 1% CWD brain homogenate (BH) and with the CWD-BH pre-incubated with quartz or Luvisolic Ae horizon for 1 or 30 weeks. After 55 weeks incubation with Chernozem and Luvisol, bound PrPCWD was not detectable by immunoblotting but remained infectious. This study shows that although recovery of PrPCWD bound to soil minerals and whole soils with time become more difficult, prion infectivity is not significantly altered. Detection of prions in soil is, therefore, not only affected by soil type but also by length of time of the prion–soil interaction.

scholarly journals Cell-surface expression of PrPC and the presence of scrapie prions in the blood of goats

2012 ◽  
Vol 93 (5) ◽  
pp. 1127-1131 ◽  
Author(s):  
Rohana P. Dassanayake ◽  
David A. Schneider ◽  
Lynn M. Herrmann-Hoesing ◽  
Thomas C. Truscott ◽  
William C. Davis ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Whole Blood ◽  
Surface Expression ◽  
Infected Sheep ◽  
Cell Surface Expression ◽  
Prion Infectivity ◽  
Infected Goat ◽  
Scrapie Infection ◽  
Relative Differences

Although host-encoded prion protein (PrPC) expression in ovine PBMCs and prion infectivity in scrapie-infected sheep blood have been demonstrated, such studies have not been reported in goats. Therefore, this study characterized cell-surface expression of PrPC on PBMC subsets derived from normal goats and sheep, by flow cytometry, and determined prion infectivity in blood from a scrapie-infected goat using a transfusion bioassay in goat kids. Cell-surface PrPC expression was detected on all subsets of goat PBMCs. The highest PrPC cell-surface expression was found in CD2+ T lymphocytes in goats. Transmission of infection was detected in all three recipients who received whole blood from a goat with classical scrapie. It was concluded that caprine PBMCs express PrPC similarly to sheep but with relative differences among PBMCs subsets, and that blood-borne infectious prions can be detected in scrapie-infected goats. Thus, similar to sheep, goat blood may be a suitable diagnostic target for the detection of scrapie infection.

scholarly journals Ex vivo mammalian prions are formed of paired double helical prion protein fibrils

Open Biology ◽  
2016 ◽  
Vol 6 (5) ◽  
pp. 160035 ◽  
Author(s):  
Cassandra Terry ◽  
Adam Wenborn ◽  
Nathalie Gros ◽  
Jessica Sells ◽  
Susan Joiner ◽  
...  
Keyword(s):  
Prion Protein ◽  
Ex Vivo ◽  
Electron Tomography ◽  
Three Dimensional ◽  
High Titre ◽  
Dimensional Structure ◽  
Structural Basis ◽  
Physical Relationship ◽  
Prion Infectivity ◽  
Mammalian Prions

Mammalian prions are hypothesized to be fibrillar or amyloid forms of prion protein (PrP), but structures observed to date have not been definitively correlated with infectivity and the three-dimensional structure of infectious prions has remained obscure. Recently, we developed novel methods to obtain exceptionally pure preparations of prions from mouse brain and showed that pathogenic PrP in these high-titre preparations is assembled into rod-like assemblies. Here, we have used precise cell culture-based prion infectivity assays to define the physical relationship between the PrP rods and prion infectivity and have used electron tomography to define their architecture. We show that infectious PrP rods isolated from multiple prion strains have a common hierarchical assembly comprising twisted pairs of short fibres with repeating substructure. The architecture of the PrP rods provides a new structural basis for understanding prion infectivity and can explain the inability to systematically generate high-titre synthetic prions from recombinant PrP.

The zoonotic potential of bat-borne coronaviruses

2020 ◽  
Vol 4 (4) ◽  
pp. 365-381
Author(s):  
Ny Anjara Fifi Ravelomanantsoa ◽  
Sarah Guth ◽  
Angelo Andrianiaina ◽  
Santino Andry ◽  
Anecia Gentles ◽  
...  
Keyword(s):  
Genetic Material ◽  
Field Research ◽  
Sympatric Species ◽  
Animal Origin ◽  
Mammalian Host ◽  
Past Century ◽  
The Past ◽  
Novel Host ◽  
Horseshoe Bat ◽  
Human Infections

Seven zoonoses — human infections of animal origin — have emerged from the Coronaviridae family in the past century, including three viruses responsible for significant human mortality (SARS-CoV, MERS-CoV, and SARS-CoV-2) in the past twenty years alone. These three viruses, in addition to two older CoV zoonoses (HCoV-229E and HCoV-NL63) are believed to be originally derived from wild bat reservoir species. We review the molecular biology of the bat-derived Alpha- and Betacoronavirus genera, highlighting features that contribute to their potential for cross-species emergence, including the use of well-conserved mammalian host cell machinery for cell entry and a unique capacity for adaptation to novel host environments after host switching. The adaptive capacity of coronaviruses largely results from their large genomes, which reduce the risk of deleterious mutational errors and facilitate range-expanding recombination events by offering heightened redundancy in essential genetic material. Large CoV genomes are made possible by the unique proofreading capacity encoded for their RNA-dependent polymerase. We find that bat-borne SARS-related coronaviruses in the subgenus Sarbecovirus, the source clade for SARS-CoV and SARS-CoV-2, present a particularly poignant pandemic threat, due to the extraordinary viral genetic diversity represented among several sympatric species of their horseshoe bat hosts. To date, Sarbecovirus surveillance has been almost entirely restricted to China. More vigorous field research efforts tracking the circulation of Sarbecoviruses specifically and Betacoronaviruses more generally is needed across a broader global range if we are to avoid future repeats of the COVID-19 pandemic.

Bacterial Shock due to Transfusion with Yersinia enterocolitica Infected Blood

1984 ◽  
Vol 16 (4) ◽  
pp. 411-412
Author(s):  
Gunnar Bjune ◽  
Tom Erik Ruud ◽  
Jan Eng
Keyword(s):  
Yersinia Enterocolitica ◽  
Infected Blood
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