scholarly journals Molecular analysis of ovine prion protein identifies similarities between BSE and an experimental isolate of natural scrapie, CH1641.

1999 ◽  
Vol 80 (1) ◽  
pp. 1-4 ◽  
Author(s):  
J Hope ◽  
S C Wood ◽  
C R Birkett ◽  
A Chong ◽  
M E Bruce ◽  
...  
Keyword(s):  
Prion Protein ◽  
Molecular Analysis ◽  
Natural Scrapie

scholarly journals Molecular analysis of ovine prion protein identifies similarities between BSE and an experimental isolate of natural scrapie, CH1641

2000 ◽  
Vol 81 (3) ◽  
pp. 851-851 ◽  
Author(s):  
J. Hope ◽  
S. C. E. R. Wood ◽  
M. E. Bruce ◽  
C. J. Bostock ◽  
A. Chong ◽  
...  
Keyword(s):  
Prion Protein ◽  
Molecular Analysis ◽  
Natural Scrapie

scholarly journals Immunohistochemical Detection and Distribution of Prion Protein in a Goat with Natural Scrapie

2003 ◽  
Vol 15 (2) ◽  
pp. 157-162 ◽  
Author(s):  
Reginald A. Valdez ◽  
Matthew J. Rock ◽  
Anne K. Anderson ◽  
Katherine I. O'Rourke
Keyword(s):  
Prion Protein ◽  
Precise Determination ◽  
Lymphoid Tissues ◽  
Large Sample Size ◽  
Parasympathetic Ganglia ◽  
Peripheral Lymph ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Natural Scrapie ◽  
Diagnostic Sensitivity And Specificity

Formalin-fixed, paraffin-embedded tissue sections from a 3-year-old female Angora goat suffering from clinical scrapie were immunostained after hydrated autoclaving using a monoclonal antibody (mAb, F99/97.6.1; IgG1) specific for a conserved epitope on the prion protein. Widespread and prominent deposition of the scrapie isoform of the prion protein (PrPSc) was observed in the brain, brainstem, spinal cord, retina, postganglionic neurons associated with parasympathetic ganglia of myenteric and submucosal plexuses, Peyer's patches, peripheral lymph nodes, and pharyngeal and palatine tonsils. The goat was homozygous for PrP alleles encoding 5 octapeptide repeat sequences in the N-terminal region of the prion protein and isoleucine at codon 142, a genotype associated with high susceptibility and short incubation times in goats. The results of this study indicate that mAb F99/97.6.1 is useful for detection of PrPSc deposition, and this is a specific and reliable immunohistochemical adjunct to histopathology for diagnosis of natural caprine scrapie, although precise determination of the diagnostic sensitivity and specificity of the assay as a diagnostic test for scrapie in goats will require examination of a sufficiently large sample size. As with ovine scrapie, prion protein is widely distributed in the central and peripheral nervous systems, gastrointestinal tract, and lymphoid tissues in natural caprine scrapie.

scholarly journals Immunohistochemical detection of prion protein in lymphoid tissues of sheep with natural scrapie.

1996 ◽  
Vol 34 (5) ◽  
pp. 1228-1231 ◽  
Author(s):  
L J van Keulen ◽  
B E Schreuder ◽  
R H Meloen ◽  
G Mooij-Harkes ◽  
M E Vromans ◽  
...  
Keyword(s):  
Prion Protein ◽  
Lymphoid Tissues ◽  
Immunohistochemical Detection ◽  
Natural Scrapie

Immumohistochemical Detection and Localization of Prion Protein in Brain Tissue of Sheep With Natural Scrapie

1995 ◽  
Vol 32 (3) ◽  
pp. 299-308 ◽  
Author(s):  
L. J. M. van Keulen ◽  
B. E. C. Schreuder ◽  
R. H. Meloen ◽  
M. Poelen-van den Berg ◽  
G. Mooij-Harkes ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Brain Tissue ◽  
Prion Protein ◽  
Medulla Oblongata ◽  
Brain Regions ◽  
Cellular Prion Protein ◽  
Immunohistochemical Method ◽  
Tissue Sections ◽  
Natural Scrapie ◽  
Detection And Localization

A converted form of the normal cellular prion protein (PrP) accumulates in the brains of sheep with scrapie. We describe an immunohistochemical method for identifying scrapie-associated PrP (PrPSc) in periodate-lysine-paraformaldehyde-fixed brain tissue, which provides adequate preservation of tissue morphology. After pretreatment of tissue sections with formic acid and hydrated autoclaving, we located PrPSc in the brains of 50 sheep with natural scrapie by use of antipeptide antisera raised against ovine PrP. No PrP was seen in 20 sheep without histopathologic signs of scrapie. PrP80 that did not stain for amyloid was present in the cytoplasm and at the cell membrane of both neurons and astrocytes. Large amounts of PrPSc were seen at the cell membrane of neurons in the medulla oblongata and pons, whereas PrPSc accumulated at the cell membrane of astrocytes of the glial limitans in all brain regions. PrPSc that stained for amyloid was located in the walls of blood vessels and perivascularly in the brains of 32 (64%) of 50 sheep, mainly in the thalamus and never in the pons or medulla oblongata. No apparent topographic relationship existed between PrPSc that stained for amyloid and PrPSc accumulation associated with neurons or astrocytes. In all scrapie-affected sheep, PrPSc was present in brain regions with vacuolation, but it could also be detected in regions with minimal or no vacuolation. We conclude that the immunohistochemical detection of PrP can be an important confirmative test in scrapie diagnosis.

scholarly journals PrPSc accumulation in placentas of ewes exposed to natural scrapie: influence of foetal PrP genotype and effect on ewe-to-lamb transmission

2002 ◽  
Vol 83 (10) ◽  
pp. 2607-2616 ◽  
Author(s):  
Olivier Andréoletti ◽  
Caroline Lacroux ◽  
Armelle Chabert ◽  
Laurent Monnereau ◽  
Guillaume Tabouret ◽  
...  
Keyword(s):  
Prion Protein ◽  
In Utero ◽  
Maternal Transmission ◽  
Trophoblastic Cells ◽  
Natural Scrapie ◽  
Temporal Accumulation

Placentas from scrapie-affected ewes are known to be infectious. Nevertheless, placenta infectivity in such ewes is not systematic. Maternal transmission to lambs is highly suspected but contamination of the foetus in utero has not been demonstrated. Using ewes from a naturally scrapie-infected flock, it was demonstrated that abnormal prion protein (PrPSc) accumulation in the placenta (i) is controlled by polymorphisms at codons 136, 154 and 171 of the foetal PrP gene and (ii) is restricted mainly to placentome foetal trophoblastic cells. In order to go deeper into the role of the placenta in scrapie transmission, the pattern of PrPSc dissemination was established in susceptible lambs (genotype VRQ/VRQ) sampled from 140 days post-insemination to the age of 4 months from either VRQ/VRQ ewes with PrPSc-positive placentas or ARR/VRQ ewes with PrPSc-negative placentas. In both VRQ/VRQ lamb groups, PrPSc spatial and temporal accumulation patterns were similar, suggesting post-natal rather than in utero contamination.

Accumulation of Pathogenic Prion Protein (PrPSc) in Nervous and Lymphoid Tissues of Sheep with Subclinical Scrapie

2003 ◽  
Vol 40 (2) ◽  
pp. 164-174 ◽  
Author(s):  
C. Ersdal ◽  
M. J. Ulvund ◽  
S. L. Benestad ◽  
M. A. Tranulis
Keyword(s):  
Lymph Node ◽  
Prion Protein ◽  
Spongiform Encephalopathy ◽  
Lymphoid Tissues ◽  
Motor Nucleus ◽  
Dorsal Motor Nucleus ◽  
The Central Nervous System ◽  
Natural Scrapie ◽  
First Time ◽  
The Brain

All sheep older than 1 year of age from a flock of the Rygja breed in which clinical scrapie was detected for the first time in two animals (4%) were examined for accumulation of pathogenic prion protein (PrPSc) by immunohistochemistry in the obex, the cerebellum, and the medial retrophayngeal lymph node. In addition, six lambs, 2–3 months old, all offspring of PrPSc-positive dams, were examined for PrPSc in the ileal Peyers' patch (IPP), the distal jejunal lymph node, the spleen, and the medial retropharyngeal lymph node (RPLN). In this flock, 35% (17/48) of the adult sheep showed accumulation of PrPSc, an eightfold increase compared with clinical disease. All positives carried susceptible PrP genotypes. Three sheep had deposits of PrPSc in the RPLN and not in the brain, suggesting that this organ, easily accessible at slaughter, is suitable for screening purposes. Two 7-year-old clinically healthy homozygous V136Q171 ewes showed sparse immunostaining in the central nervous system and may have been infected as adults. Further, two littermates, 86-days-old, showed PrPSc in the IPP. Interestingly, one of these lambs had the intermediate susceptible PrP genotype, VA136QR171. In addition to early immunolabeling in the dorsal motor nucleus of the vagal nerve, a few of the sheep had early involvement of the cerebellum. In fact, a 2-year-old sheep had sparse deposits of PrPSc in the cerebellum only. Because experimental bovine spongiform encephalopathy (BSE) in sheep seems to behave in a similar manner as natural scrapie, these results, particularly regarding spread of infectivity, may have implications for the handling of BSE should it be diagnosed in sheep.

scholarly journals Homozygosity for prion protein alleles encoding glutamine-171 renders sheep susceptible to natural scrapie.

1994 ◽  
Vol 8 (8) ◽  
pp. 959-969 ◽  
Author(s):  
D Westaway ◽  
V Zuliani ◽  
C M Cooper ◽  
M Da Costa ◽  
S Neuman ◽  
...  
Keyword(s):  
Prion Protein ◽  
Natural Scrapie

scholarly journals Different allelic effects of the codons 136 and 171 of the prion protein gene in sheep with natural scrapie

1995 ◽  
Vol 76 (8) ◽  
pp. 2097-2101 ◽  
Author(s):  
C. Clouscard ◽  
P. Beaudry ◽  
J. M. Elsen ◽  
D. Milan ◽  
M. Dussaucy ◽  
...  
Keyword(s):  
Prion Protein ◽  
Protein Gene ◽  
Prion Protein Gene ◽  
Natural Scrapie

scholarly journals Proteinase K-sensitive disease-associated ovine prion protein revealed by conformation-dependent immunoassay

2006 ◽  
Vol 401 (2) ◽  
pp. 475-483 ◽  
Author(s):  
Alana M. Thackray ◽  
Lee Hopkins ◽  
Raymond Bujdoso
Keyword(s):  
Prion Protein ◽  
Phosphotungstic Acid ◽  
Limited Proteolysis ◽  
Proteinase K ◽  
Genotypic Differences ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Natural Scrapie ◽  
Proteinase K Treatment ◽  
Sheep Scrapie

PrPSc [abnormal disease-specific conformation of PrP (prion-related protein)] accumulates in prion-affected individuals in the form of amorphous aggregates. Limited proteolysis of PrPSc results in a protease-resistant core of PrPSc of molecular mass of 27–30 kDa (PrP27–30). Aggregated forms of PrP co-purify with prion infectivity, although infectivity does not always correlate with the presence of PrP27–30. This suggests that discrimination between PrPC (normal cellular PrP) and PrPSc by proteolysis may underestimate the repertoire and quantity of PrPSc subtypes. We have developed a CDI (conformation-dependent immunoassay) utilizing time-resolved fluorescence to study the conformers of disease-associated PrP in natural cases of sheep scrapie, without using PK (proteinase K) treatment to discriminate between PrPC and PrPSc. The capture-detector CDI utilizes N-terminal- and C-terminal-specific anti-PrP monoclonal antibodies that recognize regions of the prion protein differentially buried or exposed depending on the extent of denaturation of the molecule. PrPSc was precipitated from scrapie-infected brain stem and cerebellum tissue following sarkosyl extraction, with or without the use of sodium phosphotungstic acid, and native and denatured PrPSc detected by CDI. PrPSc was detectable in brain tissue from homozygous VRQ (V136 R154 Q171) and ARQ (A136 R154 Q171) scrapie-infected sheep brains. The highest levels of PrPSc were found in homozygous VRQ scrapie-infected brains. The quantity of PrPSc was significantly reduced, up to 90% in some cases, when samples were treated with PK prior to the CDI. Collectively, our results show that the level of PrPSc in brain samples from cases of natural scrapie display genotypic differences and that a significant amount of this material is PK-sensitive.

scholarly journals Comparative Molecular Analysis of the Abnormal Prion Protein in Field Scrapie Cases and Experimental Bovine Spongiform Encephalopathy in Sheep by Use of Western Blotting and Immunohistochemical Methods

2004 ◽  
Vol 78 (7) ◽  
pp. 3654-3662 ◽  
Author(s):  
Stéphane Lezmi ◽  
Stuart Martin ◽  
Stéphanie Simon ◽  
Emmanuel Comoy ◽  
Anna Bencsik ◽  
...  
Keyword(s):  
Bovine Spongiform Encephalopathy ◽  
Prion Protein ◽  
Western Blotting ◽  
Molecular Analysis ◽  
Infected Sheep ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spongiform Encephalopathy ◽  
Protease Cleavage ◽  
Two Samples ◽  
Immunohistochemical Methods

ABSTRACT Since the appearance of bovine spongiform encephalopathy (BSE) in cattle and its linkage with the human variant of Creutzfeldt-Jakob disease, the possible spread of this agent to sheep flocks has been of concern as a potential new source of contamination. Molecular analysis of the protease cleavage of the abnormal prion protein (PrP), by Western blotting (PrPres) or by immunohistochemical methods (PrPd), has shown some potential to distinguish BSE and scrapie in sheep. Using a newly developed enzyme-linked immunosorbent assay, we identified 18 infected sheep in which PrPres showed an increased sensitivity to proteinase K digestion. When analyzed by Western blotting, two of them showed a low molecular mass of unglycosylated PrPres as found in BSE-infected sheep, in contrast to other naturally infected sheep. A decrease of the labeling by P4 monoclonal antibody, which recognizes an epitope close to the protease cleavage site, was also found by Western blotting in the former two samples, but this was less marked than in BSE-infected sheep. These two samples, and all of the other natural scrapie cases studied, were clearly distinguishable from those from sheep inoculated with the BSE agent from either French or British cattle by immunohistochemical analysis of PrPd labeling in the brain and lymphoid tissues. Final characterization of the strain involved in these samples will require analysis of the features of the disease following infection of mice, but our data already emphasize the need to use the different available methods to define the molecular properties of abnormal PrP and its possible similarities with the BSE agent.

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