Volume 25, Number 1—January 2019
Research
Variable Protease-Sensitive Prionopathy Transmission to Bank Voles
Romolo Nonno1, Silvio Notari1, Michele Angelo Di Bari, Ignazio Cali, Laura Pirisinu, Claudia d’Agostino, Laura Cracco, Diane Kofskey, Ilaria Vanni, Jody Lavrich, Piero Parchi, Umberto AgrimiComments to Author , and Pierluigi GambettiComments to Author
Author affiliations: Istituto Superiore di Sanità, Rome, Italy (R. Nonno, M.A. Di Bari, L. Pirisinu, C. d’Agostino, I. Vanni, U. Agrimi); Case Western Reserve University, Cleveland, Ohio, USA (S. Notari, I. Cali, L. Cracco, D. Kofskey, J. Lavrich, P. Gambetti); University of Bologna, Bologna, Italy (P. Parchi); Istituto di Ricovero e Cura a Carattere Scientifico, Bologna (P. Parchi)
Abstract
Variably protease-sensitive prionopathy (VPSPr), a recently described human sporadic prion disease, features a protease-resistant, disease-related prion protein (resPrPD) displaying 5 fragments reminiscent of Gerstmann-Sträussler-Scheinker disease. Experimental VPSPr transmission to human PrP–expressing transgenic mice, although replication of the VPSPr resPrPD profile succeeded, has been incomplete because of second passage failure. We bioassayed VPSPr in bank voles, which are susceptible to human prion strains. Transmission was complete; first-passage attack rates were 5%–35%, and second-passage rates reached 100% and survival times were 50% shorter. We observed 3 distinct phenotypes and resPrPD profiles; 2 imitated sporadic Creutzfeldt-Jakob disease resPrPD, and 1 resembled Gerstmann-Sträussler-Scheinker disease resPrPD. The first 2 phenotypes may be related to the presence of minor PrPD components in VPSPr. Full VPSPr transmission confirms permissiveness of bank voles to human prions and suggests that bank vole PrP may efficiently reveal an underrepresented native strain but does not replicate the complex VPSPr PrPD profile.
Sporadic prion diseases are classified according to phenotype as well as the pairing of the prion protein (PrP) genotype at the methionine (M)/valine (V) polymorphic codon 129 and the conformational characteristics of the abnormal or disease-associated PrP (PrPD). These characteristics include electrophoretic mobility and the ratio of the PrPD fragments that are resistant to proteinase K (PK) digestion (AppendixTable 1) (1). According to these criteria, the 3 major types of sporadic prion disease are sporadic Creutzfeldt-Jakob disease (sCJD), sporadic fatal insomnia, and variably protease-sensitive prionopathy (VPSPr) (2–5).
VPSPr was first reported in 2008 and further defined in 2010 (6–8) as a sporadic prion disease distinct from sCJD. Since then, 37 cases have been reported, consistent with a prevalence rate of 1%–2% for all sporadic prion diseases (8). Similar to sCJD, VPSPr targets all 3 PrP genotypes. However, the prevalence of the 3 genotypes at codon 129 (MM, MV, and VV) greatly differs, indeed is almost inverted, in the 2 diseases: homozygosity VV is the most common (65%) genotype in VPSPr and the least common (16%) in sCJD (2,9). Furthermore, at variance with sCJD, in which the 129 genotype is a determinant of disease phenotype and PrPD characteristics, the 129 genotype influence on phenotype, although present, is subtle (3,7,8). These differences point to a distinct role of the 129 genotype as a risk factor and imply that the etiologic-pathogenetic mechanisms of the 2 diseases differ.
Although the histopathology of VPSPr is distinct (e.g., spongiform degeneration, frequent presence of PrP microplaques, and a recognizable PrPD immunostaining pattern), the hallmarks of VPSPr are the characteristics of its PrPD. In contrast to virtually all other sporadic human prion diseases, in which PK-resistant PrPD (resPrPD) electrophoretically separates into 3 major bands, VPSPr resPrPD characteristically separates into 5 bands. Furthermore, although the 3 bands of resPrPD are all cleaved by PK exclusively at the N terminus and separate according to the presence of 2, 1, or 0 sugar moieties, VPSPr resPrPD bands include only the monoglycosylated and unglycosylated forms, which are cleaved either only at the N terminus or at both the N- and C-termini. Thus, the C-terminus–truncated resPrPD lacks the GPI (glycosylphosphatidylinositol) anchor. Additional variances concerning immunoreactivity characteristics, ratios of PK-resistant, and PK-sensitive PrPD species and conformational properties including aggregate size, have also been observed (6–8). These distinctive properties point to VPSPr PrPD as a prion strain different from those of other sporadic prion diseases. However, the VPSPr prion shares the multiplicity of the resPrPD electrophoretic bands with prions from a subset of inherited prion diseases referred to as Gerstmann-Sträussler-Scheinker disease (GSS), prompting the suggestion that VPSPr is the sporadic form of GSS (7,10). Furthermore, the presence of small amounts of sCJD-like 3-band resPrPD has also been signaled in VPSPr (6,11,12).
Disease transmission to receptive hosts is a valuable way to further define the characteristics of strains associated with prion diseases. VPSPr has been experimentally transmitted to 3 lines of transgenic mice expressing normal PrP or cellular human PrP (PrPC), harboring residue M, V, or MV at residue 129 (13,14). Data in all experiments were essentially similar. Inoculated mice remained asymptomatic, but half showed focal PrPD plaques with minimal spongiform degeneration, and PrPD mimicking the electrophoretic profile of the native PrPD on immunoblot was demonstrated in about one third of the inoculated mice. No transmission was observed at second passage.
The bank vole, a small rodent resembling the mouse with which it shares the entire sequence of normal PrP or PrPC except for 8 aa, but whose sequence differs from human PrPC by 15 aa, has recently emerged as a particularly permissive host. Bank voles and transgenic mice expressing bank vole PrPC have been successfully infected after challenge with human and animal prion diseases that are hard to transmit even to recipients expressing homologous PrPC (15–18).
We studied transmission of VPSPr from patients with MM, MV, and VV codon 129 genotypes to bank voles harboring either the PrP genotype 109M (bv109M) or 109I (bv109I). Although the attack rate was generally low at first passage, it consistently raised to 100% at second passage, when survival times also decreased on average by >50%. We identified 3 PrPD isoforms with the characteristics of distinct strains in the affected bank voles.
Materials and Methods The inocula used in the first passage were brain homogenates from 7 persons with a definitive diagnosis of VPSPr: 2 with genotypes 129MM, 3 with 129MV, and 2 with 129VV. Homogenate was inoculated into the cerebrum of 205 bank voles according to previously described procedures (16). The bank vole brains were processed for histopathology, immunohistochemistry, lesion profiles, and paraffin-embedded tissue (PET) blots according to previously reported procedures (15). Western blot was performed according to Notari et al. (19). The insoluble fraction was prepared according to previously described procedures (20). Preparation of monoclonal antibodies is described in the Appendix. Statistical significance was determined by 1-way analysis of variance, followed by the Tukey multiple comparison test.
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Results Transmission Characteristics At first passage, attack rates of VPSPr were 35% (29/82) in bv109I and 5% (3/59) in bv109M (Table 1; Appendix Table 2). The 2 bank vole genotypes diverged as to disease transmission in 2 ways. First, all VPSPr 129 genotypes were transmitted to bv109I, but bv109M were not susceptible to VPSPr-VV. Second, bv109I propagated 3 distinct histopathologic phenotypes and matching PrPD types (hereafter identified as T1, T2, and T3), but bv109M replicated the T1 phenotype exclusively. A more detailed analysis in bv109I, although limited by the low number of animals in each subset, suggested a lower attack rate for VPSPr-VV, the most common form of human VPSPr, compared with the MM and MV genotypes and a prevalence for T3 that was 11% higher than that for T1 and 106% higher than that for T2 (Table 2). Overall survival times were 575 days postinoculation (dpi) for bv109I and 411 dpi for bv109M. However, when we considered only the bank voles associated with the T1 phenotype, because bv109M were exclusively associated with T1, the dpi difference became smaller: 490 dpi for bv109I and 411 dpi for bv109M (Tables 1, 2). As for survival times related to T1–T3 phenotypes and VPSPr genotypes, the survival times for T2 were nearly twice those for T1 and T3 (Table 2).
Second passage in bv109I was invariably characterized by a 100% attack rate, a 40%–61% decrease in survival times, and conservation of the original phenotype (Table 1). A similar trend was observed for bv109M.
Histopathology and Immunohistochemistry
Phenotype T1 featured finely vacuolated spongiform degeneration often involving the entire thickness of the neocortex, including the molecular layer but occasionally also showing a laminar distribution (Figure 1). On second passage, the spongiform degeneration appeared to be more widespread, also affecting the hippocampus and subcortical structures such as basal nuclei, thalamus, and superior colliculi but not the cerebellum. PrP immunohistochemistry demonstrated punctate deposits often co-distributed with spongiform degeneration (Figure 1, column T1, row ii). At second passage, T1 features did not differ significantly between bv109M and bv109I. Furthermore, T1 also resembled the histopathologic phenotype shown by bv109M and bv109I after inoculation with sCJDMM1 or sCJDMV1, respectively (Figures 2, 3; Appendix Figure 1)
In phenotype T2, spongiform degeneration affected predominantly subcortical structures over neocortical regions, especially the hypothalamus with the apparent exclusion of the mammillary bodies (Figure 1, column T2, row iii). PrP immunohistochemistry showed granular deposits occasionally resembling mini plaquelike formations rather than the punctate deposits of the T1 phenotype (Figure 1, column T2, row iv).
Phenotype T3 was characterized by the paucity of spongiform degeneration in the cerebral neocortex and subcortical gray matter structures; spongiform degeneration was often prominent in the regions of the hemispheric white matter lying above the hippocampus and in the corpus callosum, where parenchyma was occasionally disorganized with glial reaction. PrP immunostaining was mostly limited to those regions where it often aggregated in confluent plaque-like deposits but not well-formed plaques (Figure 1, column T3, row v, and column Tc, row vi). No remarkable differences were detected between first and second passages. Overall, the T3 histopathologic phenotype resembled that shown by bv109I after inoculation with brain homogenates from some GSS subtypes (16).
It is noteworthy that the T1–T3 phenotypes were never observed to coexist in 1 animal, although distinct phenotypes were often observed in bank voles receiving the same inoculum. Although all 3 phenotypes occurred after inoculation with VPSPr-MM or -MV, the sole phenotype associated with VPSPr-VV inoculation was T3 (Table 1).
Lesion Profiles and PET Blots Profiles of spongiform degeneration as a function of lesion severity and brain anatomic location confirmed the distinctive characteristics of the T1–T3 phenotypes (Figure 2, panel A; Appendix Figure 1). The T1 spongiform degeneration profile in bv109I did not differ significantly from that of bv109M; both mirrored the profiles of bv109I inoculated with sCJDMV1 and bv109M inoculated with sCJDMM1 brain homogenate (Figure 2, panel B; AppendixFigure 1).
The PET blot patterns of brain PrPD distribution were also quite distinct in the 3 phenotypes and, overall, reproduced the spongiform degeneration distribution (Figures 2, 3). In T1, PrPD was well represented in selected regions including cerebral neocortex and hippocampus, basal nuclei, thalamus, superior colliculi, geniculate nuclei, and substantia nigra but not in the cerebellum and lower brain stem. No significant variations were detected between PrPD distributions at first and second passages (data not shown). PrPD distributions were also similar in bv109I and bv109M inoculated with classic sCJDMV1 and sCJDMM1 prions, respectively (Figure 3, panel B). In the T2 phenotype, PrPD appeared to be present in moderate and uniform amounts in several anatomic regions such as neocortex and hippocampus, thalamus, and superior colliculi (Figure 3). The T3 phenotype was characterized by the striking presence of PrPD in hippocampus and white matter structures (Figure 3).
PrPD Characterization
Immunoblot analysis confirmed the presence of 3 distinct resPrPD electrophoretic profiles that matched the 3 histopathologic phenotypes. When probed with antibodies 9A2 and 12B2, resPrPD associated with the T1 phenotype populated 3 bands of ≈32, 26, and 21 kDa, representing the 3 resPrPD glycoforms, and by a fragment of 7 kDa (Figure 4). An additional C-terminal fragment of ≈13 kDa, possibly homologous to the human C-terminal fragment 12/13 (20), was detected by the C-terminal antibody SAF84 (Figure 4). Glycoform ratios showed a comparable representation of the diglycosylated and monoglycosylated forms of resPrPD (Figure 5; Appendix Figure 2). The electrophoretic profile and glycoform ratios of resPrPD T1 conformer were indistinguishable from those of resPrPD observed in bank voles inoculated with sCJDMM1 or sCJDMV1 prions, used as controls for human resPrPD type 1 in bank voles (Figures 4, 5; Appendix Figure 2; data not shown).
The resPrPD profile associated with the T2 phenotype showed 3 bands of ≈30, 24, and 19 kDa (i.e., all that had an ≈2-kDa faster electrophoretic mobility than the corresponding bands of resPrPD T1) (Figure 4). The 7-kDa fragment was not detected in T2 (Figure 4). In contrast to T1, the T2 glycoform ratio was characterized by the unambiguous predominance of the monoglycosylated component (Figure 5). In summary, bank vole resPrPD T2 differed from the T1 conformer by overall 2-kDa faster mobility, the absence of the 7-kDa fragment, and marked predominance of the monoglycoform. The striking feature of the resPrPD associated with the T3 phenotype was the predominant presence of the 7-kDa fragment detected by 9A2 and 12B2 but not by SAF84, demonstrating its internal origin and the absence of glycosylation sites (Figure 4).
Additional divergent features emerged when amounts of totPrPD (i.e., PK-sensitive plus resPrPD fractions) were assessed as percentages of total PrP, comprising PrPC and totPrPD (Figure 6). A significantly larger component of totPrPD was resPrPD in T1 than in T2 (81% vs. 33%); totPrPD fractions were similar (93% for T1, 91% for T2). T3 differed significantly: totPrPD accounted for 8% and resPrPD accounted for 0.2% of total PrP (Figure 6; Appendix Figure 3).
Discussion The permissiveness of bank vole PrPC is well known (15,16,18,21–27); it is exemplified by the observation that, despite the mere 8-aa PrPC divergence between bank voles and mice, a variety of human and animal prion diseases not transmissible to mice are infectious to bank voles and transgenic mice expressing bank vole PrP (15,16,18,22,24). Conversely, the 15-aa difference from the human PrPC primary sequence does not impede the efficient transmission of a wealth of sporadic and inherited human prion diseases (15,16). This striking permissiveness has been attributed to the presence of several asparagine and glutamine residues in and around the β2–α2 loop that would result in a PrPC conformation compatible with the conformations of a large number of PrPD strains (21). Furthermore, the polymorphism at bank vole codon 109 adds further complexity to the interaction with exogenous strains (18,28).
We undertook systematic transmission of VPSPr brain homogenates to bv109M and bv109I after failure to consistently transmit VPSPr to humanized transgenic mice. Overall, transmission was favored by the 109I genotype, which propagated all 3 VPSPr 129 genotypes while bv109M failed to transmit VPSPr-VV. However, at first passage in bv109I, the mean attack rate (35%) was fairly low and the mean survival time (575 dpi) quite extended (Table 1). These conditions changed at second passage, when the attack rate became 100% in all transmission experiments and survival times decreased on average by 53% (Table 1). These findings point to the existence of a substantial barrier at first passage, which, judging from the 100% attack rate, is probably largely diminished or vanished at second passage. In view of the aforementioned easy transmissibility of other human prion diseases, the barrier appears to be conformational rather than caused by species-related variations in amino acid sequence of PrPC (15,16); the barrier might be associated with the misfolding of VPSPr PrPD, which may be peculiar because after PK digestion it results in an array of highly heterogeneous fragments and apparently the failure to convert one of the glycoforms (6,7). Similarly, the clear effect of the genotype at codon 129 on the attack rate (which was 3–4 times lower for bank voles inoculated with VPSPr-VV prions compared with VPSPr-MM and -MV), along with the lack of transmission of VPSPr-VV to bv109M but not to bv109I, points to conformational differences between PrPD species associated with the 129 genotypes in VPSPr (16). This notion is further supported by previous data showing higher PK sensitivity (7) and conformational stability of PrPD (29) in VPSPr-VV compared with VPSPr-MM and -MV.
The comparative study of VPSPr bioassay in bank voles and humanized transgenic mice revealed substantial differences. VPSPr-challenged mice invariably remained asymptomatic, and all histologically positive mice failed to transmit at second passage. Furthermore, the VPSPr-MV subtype was never transmitted to mice 129M or 129V, and the general attack rate (assessed histopathologically) was low (54%); resPrPD was demonstrated in only 34% of the challenged mice despite the 2–8 times normal levels of PrP expression for most mice (13). However, in contrast to bank voles, positive mice generated a resPrPD conformer very similar to that of VPSPr for electrophoretic profile, glycosylation pattern, and antibody immunoreactivity, although it exhibited higher protease resistance.
Data from a previous study of transmission to humanized transgenic mice and bv109M of an sCJDMV variant with atypical glycoform profile (CJD-MVAG) partially resembled ours (17). Challenged transgenic mice remained asymptomatic and negative at neuropathologic examination, but 22% of them reproduced the original resPrPD electrophoretic profile and glycotype of the inoculum. In contrast to humanized transgenic mice, bank voles had full-blown disease develop featuring 3, although partially merging, histopathologic phenotypes along with 3 distinct resPrPD conformers, none of which mimicked the profile and glycotype of the inoculum (17). Remarkably, the glycoform variation of sCJDMVAG resembles that of VPSPr because both resPrPD species lack the diglycosylated isoform, implicating this variation as one of the possible causes of bank vole failure to accurately replicate exogenous PrPD (17).
Three subtypes of GSS (which VPSPr resembles in terms of the ladder-like electrophoretic profile and the sensitivity to PK of resPrPD), have also recently been transmitted to bank voles and 1 GSS subtype to humanized transgenic mice (16,30). Despite the well-known difficulty of transmitting GSS to rodents, bank voles challenged with 2 major GSS subtypes, associated with PrP mutations A117V and F198S (GSSA117V, GSSF198S) showed no evidence of species or mutation barrier. Transmission was comparatively more difficult with the third GSSP102L subtype, in which resPrPD displays 2 sets of fragments: either the 8-kDa fragment associated with the 30–21 kDa glycoform triplet (31,32) or the 8-kDa fragment alone. After inoculation, the 2-fragment set was never replicated, and the ≈8-kDa fragment alone occasionally was inaccurately reproduced as a 7-kDa fragment (16,28). To date, only GSSA117V has been transmitted to 2 lines of transgenic mice expressing human PrPD harboring the A117V transition (30). Although transmission features diverged in the 2 lines, both seemed to reproduce the 7-kDa fragment that is the only strongly resPrPD fragment in this disease.
Combined, these experiments indicate that PrPC characteristics, and possibly other host factors (25), enable bank voles to be more permissive hosts (despite the species barrier) than transgenic mice expressing conspecific PrPC, confirming the empirical aspect of the species barrier. However, bank vole PrPC can hardly reproduce faithfully complex features of human atypical prion isolates, a task that may require PrPC from the same species.
A remarkable finding of this study is the occurrence of 3 well-defined histopathologic phenotypes (T1–T3), which displayed discrete PrPD brain distribution and were linked to PrPD conformers easily distinguishable by electrophoretic profile and glycosylation characteristics. The 3 phenotypes also differed by mean survival times at first and second passages. Remarkably, the T1–T3 phenotypes were often generated by the same inoculum but never co-occurred in the same bank vole. Combined, these features define the T1–T3 PrPD conformers as distinct strains, raising the issue of their origin. Both histopathologic and resPrPD electrophoretic characteristics of the T1 phenotype are essentially indistinguishable from those of bank voles inoculated with sCJDMV1. Data on transmission of sCJDMM2, available only for bv109M, show that the electrophoretic profile of the newly formed resPrPD matches the T2 resPrPD of this study (15). Although the T1 and T2 representations of totPrPD and resPrPD are not known in bank voles inoculated with sCJDMM1 and sCJDMM2 prions, the values we observed after VPSPr inoculation are comparable to those reported for the original sCJD, in which totPrPD and resPrPD reportedly accounted for 53.5% and 48.2% of total PrP in sCJDMM1 (6; L. Cracco et al., unpub. data). Therefore, transmission to bank voles suggests that VPSPr PrPD T1 and T2 are related to human PrPD types 1 and 2, respectively. In contrast, phenotype T3 is the most divergent, especially for spongiform degeneration and PrPD deposition, mostly limited to white matter regions, and electrophoretic profile, where resPrPD recovered as a band of 7 kDa was the major component shared with the complex pattern of VPSPr resPrPD. The T3 histopathologic phenotypes including the PrP immunostaining pattern matched also the bank vole phenotype of GSSA117V and GSSP102L associated with the 8-kDa fragment only (16). The exceedingly low representation of the totPrPD and resPrPD components of total PrP in T3 is reminiscent of the corresponding data reported in VPSPr-VV, in which totPrPD accounted for 3.4% and resPrPD for 0.83% of total PrP (6). The marked underrepresentation of totPrPD and resPrPD in T3 is especially puzzling considering that attack rate and survival time are not very different from those of T2 and T1, respectively. The apparent relative high efficiency of T3 might be explained by the high representation of oligomers (36). Alternatively, the T3 underrepresentation of totPrPD relative to total PrP might reflect the lack of PrPC down-regulation by T3 compared with T1 and T2, which would result in the relative increase of the total PrP pool (33,34).
A mechanism put forward for the lack of fidelity in cross-species transmission of the prion strain (25,35–37) is based on evidence that the dominant strain is selected from an array of strains that persist as substrains. In cross-species transmissions, substrains may be selected over the dominant strain (38–40). In the context of VPSPr, this mechanism is particularly intriguing, given that small quantities of PrPD conformers with electrophoretic mobilities similar to those of human PrPD types 1 were originally observed in a few cases by Gambetti et al (6); more recently, the presence of PrPD type 2 in VPSPr, mostly in subcortical nuclei and in cerebellum, has been reported (11,12). These 2 components would be propagated faithfully in T1 and T2, and T3, which consistently shares only the 7-kDa fragment with the VPSPr resPrPD, might represent the selective amplification of this GSS-like VPSPr component, perhaps because of the unsuccessful attempt to fully reproduce the dominant strain associated with this disease. We and others have occasionally observed an underrepresented 7-kDa fragment in sCJDMM1 (41; S. Notari, P. Gambetti, P. Parchi, unpub. data). Thus, it is tempting to speculate that the 7-kDa fragment observed in bank voles inoculated with sCJDMM1 and sCJDMV1 prions is related to the presence and possibly the infectivity of such fragment in the sCJDMM(MV)1 subtype.
In conclusion, on the basis of the first full transmission of VPSPr, our study confirms the permissiveness of bank voles to human prion diseases and suggests that bank voles are competent to reveal minor strain variants in prion diseases, such as resPrPD types 1 and 2 reported in VPSPr and, possibly, the ≈7-kDa fragment observed in sCJDMM1 and sCJDMV1. However, our study also underscores the limited competence of bank vole PrPC to faithfully reproduce the multiband profile of VPSPr resPrPD that probably reflects the complex conformation of the prion seed in this disease.
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Dr. Nonno is a research scientist in the Emerging Zoonosis Unit at the Istituto Superiore di Sanità in Rome, Italy; his primary research interests include prion strain characterization and the zoonotic potential of animal prion diseases. Dr. Notari is an instructor in the Department of Pathology at Case Western Reserve University, Cleveland, Ohio, USA; his research focuses mainly on prion diseases, particularly prion molecular characteristics and infectivity.
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Acknowledgments
snip...end
MONDAY, NOVEMBER 26, 2018
***> Sporadic Creutzfeldt-Jakob Disease in a Woman Married Into a Gerstmann-Sträussler-Scheinker Family: An Investigation of Prions Transmission via Microchimerism
Tuesday, March 20, 2018
***> Variably protease-sensitive prionopathy (VPSPr), sporadic creutzfeldt jakob disease sCJD, the same disease, what if?
Monday, August 9, 2010
Variably protease-sensitive prionopathy: A new sporadic disease of the prion protein or just more Prionbaloney ?
DEEP THROAT TO TSS 2000-2001 (take these old snips of emails with how ever many grains of salt you wish. ...tss)
The most frightening thing I have read all day is the report of Gambetti's finding of a new strain of sporadic cjd in young people...Dear God, what in the name of all that is holy is that!!! If the US has different strains of scrapie.....why???? than the UK...then would the same mechanisms that make different strains of scrapie here make different strains of BSE...if the patterns are different in sheep and mice for scrapie.....could not the BSE be different in the cattle, in the mink, in the humans.......
I really think the slides or tissues and everything from these young people with the new strain of sporadic cjd should be put up to be analyzed by many, many experts in cjd........bse.....scrapie Scrape the damn slide and put it into mice.....wait.....chop up the mouse brain and and spinal cord........put into some more mice.....dammit amplify the thing and start the damned research.....This is NOT rocket science...we need to use what we know and get off our butts and move....the whining about how long everything takes.....well it takes a whole lot longer if you whine for a year and then start the research!!!
Not sure where I read this but it was a recent press release or something like that: I thought I would fall out of my chair when I read about how there was no worry about infectivity from a histopath slide or tissues because they are preserved in formic acid, or formalin or formaldehyde.....for God's sake........ Ask any pathologist in the UK what the brain tissues in the formalin looks like after a year.......it is a big fat sponge...the agent continues to eat the brain ......you can't make slides anymore because the agent has never stopped........and the old slides that are stained with Hemolysin and Eosin......they get holier and holier and degenerate and continue...what you looked at 6 months ago is not there........Gambetti better be photographing every damned thing he is looking at.....
Okay, you need to know. You don't need to pass it on as nothing will come of it and there is not a damned thing anyone can do about it. Don't even hint at it as it will be denied and laughed at.......... USDA is gonna do as little as possible until there is actually a human case in the USA of the nvcjd........if you want to move this thing along and shake the earth....then we gotta get the victims families to make sure whoever is doing the autopsy is credible, trustworthy, and a saint with the courage of Joan of Arc........I am not kidding!!!! so, unless we get a human death from EXACTLY the same form with EXACTLY the same histopath lesions as seen in the UK nvcjd........forget any action........it is ALL gonna be sporadic!!!
And, if there is a case.......there is gonna be every effort to link it to international travel, international food, etc. etc. etc. etc. etc. They will go so far as to find out if a sex partner had ever traveled to the UK/europe, etc. etc. .... It is gonna be a long, lonely, dangerous twisted journey to the truth. They have all the cards, all the money, and are willing to threaten and carry out those threats....and this may be their biggest downfall...
Thanks as always for your help. Recently had a very startling revelation from a rather senior person in government here..........knocked me out of my chair........you must keep pushing. If I was a power person....I would be demanding that there be a least a million bovine tested as soon as possible and aggressively seeking this disease. The big players are coming out of the woodwork as there is money to be made!!! In short: "FIRE AT WILL"!!! for the very dumb....who's "will"! "Will be the burden to bare if there is any coverup!"
again it was said years ago and it should be taken seriously....BSE will NEVER be found in the US! As for the BSE conference call...I think you did a great service to freedom of information and making some people feign integrity...I find it scary to see that most of the "experts" are employed by the federal government or are supported on the "teat" of federal funds. A scary picture! I hope there is a confidential panel organized by the new government to really investigate this thing.
USA EMERGENCY 50 STATE BSE CONFERENCE CALL
-----Original Message-----
From: Terry Singeltary <flounder9@verizon.net>
To: bse-l <bse-l@lists.aegee.org>
Cc: cjd-l <cjd-l@lists.aegee.org>; cjdvoice <cjdvoice@yahoogroups.com>; bloodcjd <bloodcjd@yahoogroups.com>
Sent: Tue, Mar 20, 2018 1:00 pm
Subject: Variably protease-sensitive prionopathy (VPSPr), sporadic creutzfeldt jakob disease sCJD, the same disease, what if?
Emergence of Novel Strains
From: Terry Singeltary <flounder9@verizon.net>
To: bse-l <bse-l@lists.aegee.org>
Cc: cjd-l <cjd-l@lists.aegee.org>; cjdvoice <cjdvoice@yahoogroups.com>; bloodcjd <bloodcjd@yahoogroups.com>
Sent: Tue, Mar 20, 2018 1:00 pm
Subject: Variably protease-sensitive prionopathy (VPSPr), sporadic creutzfeldt jakob disease sCJD, the same disease, what if?
Variably protease-sensitive prionopathy (VPSPr), sporadic creutzfeldt jakob disease sCJD, the same disease, what if?
VPSPr, and the quest/rush to have a disease named after you, even if it's the same disease as sporadic CJD?
Variant CJD
Abigail B Diack, Mark W Head, Sandra McCutcheon, Aileen Boyle, Richard Knight, James W Ironside, Jean C Manson & Robert G Will To cite this article: Abigail B Diack, Mark W Head, Sandra McCutcheon, Aileen Boyle, Richard Knight, James W Ironside, Jean C Manson & Robert G Will (2014) Variant CJD, Prion, 8:4, 286-295, DOI: 10.4161/pri.29237 To link to this article: https://doi.org/10.4161/pri.29237
Variant CJD
18 years of research and surveillance
Abigail B Diack1,y , Mark W Head2,y , Sandra McCutcheon1 , Aileen Boyle1 , Richard Knight2 , James W Ironside2 , Jean C Manson1,y, *, and Robert G Will2,y 1 The Roslin Institute and R(D)SVS; University of Edinburgh; Easter Bush; Midlothian, Scotland, UK; 2 National CJD Research & Surveillance Unit; School of Clinical Sciences; University of Edinburgh; Western General Hospital; Edinburgh, Scotland, UK y These authors contributed equally to this work. Keywords: prion, variant Creutzfeldt–Jakob disease, transmissible spongiform encephalopathy, prion protein Abbreviations: BSE, bovine spongiform encephalopathy; CWD, chronic wasting disease; GSS, Gerstman–Str€aussler–Scheinker disease; M, methionine; PPS, pentosan polysulphate; PrPres, protease-resistant prion protein; PrPSc, abnormal prion protein; QuIC, quaking-induced conversion; TSE, transmissible spongiform encephalopathy; V, valine; vCJD, variant Creutzfeldt–Jakob disease; VPSPr, variably protease-sensitive prionopathy
It is now 18 years since the first identification of a case of vCJD in the UK. Since that time, there has been much speculation over how vCJD might impact human health. To date there have been 177 case reports in the UK and a further 51 cases worldwide in 11 different countries. Since establishing that BSE and vCJD are of the same strain of agent, we have also shown that there is broad similarity between UK and non-UK vCJD cases on first passage to mice. Transgenic mouse studies have indicated that all codon 129 genotypes are susceptible to vCJD and that genotype may influence whether disease appears in a clinical or asymptomatic form, supported by the appearance of the first case of potential asymptomatic vCJD infection in a PRNP 129MV patient. Following evidence of blood transfusion as a route of transmission, we have ascertained that all blood components and leucoreduced blood in a sheep model of vCJD have the ability to transmit disease. Importantly, we recently established that a PRNP 129MV patient blood recipient with an asymptomatic infection and limited PrPSc deposition in the spleen could readily transmit disease into mice, demonstrating the potential for peripheral infection in the absence of clinical disease. This, along with the recent appendix survey which identified 16 positive appendices in a study of 32 441 cases, underlines the importance of continued CJD surveillance and maintaining control measures already in place to protect human health.
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Identification of novel strains involves veterinary and medical vigilance, but it also requires a proper and full characterization of known prion agents. While the deployment of wild-type mouse panels, transgenic mice, and non-human primates all rapidly concluded that vCJD was a novel human prion strain related to BSE (see above), determining how many distinct human prion strains there are has proved surprisingly difficult, especially for sCJD. Transmission studies in humanised transgenic mice and non-human primates point to 4 major groups within sporadic, iatrogenic CJD, Kuru, and some genetic CJD cases, termed M1, V1, M2, and V2.22,106 Sporadic fatal insomnia and fatal familial insomnia (FFI) together may represent a sixth strain107 and 2 further transmissible phenotypes can be derived from GSS disease: one involving a transmissible amyloid phenotype, the other a fully transmissible spongiform encephalopathy.108,109 The transmission properties of PrP cerebral amyloid angiopathy and Variably ProteaseSensitive Prionopathy (VPSPr) remain to be reported. The relationship between human disease phenotypes, agent strain, and prion biochemistry is further complicated by the now widely recognized phenomenon of distinct PrPres type co-occurrence in the sCJD, vCJD and VPSPr brain.15,24,110
Surveillance for BSE in cattle, sheep, and goats has identified new (or newly discovered) animal prion diseases including atypical scrapie in sheep and so called H- and L-type BSE in cattle. These along with chronic wasting disease (CWD) in deer and elk represent a potential zoonotic risk to human health that is hard to quantify. While Wilson et al.111 have shown no transmission of CWD, BASE, H-type BSE, and atypical scrapie to mice expressing wild-type levels of human PrP, Kong et al.112 demonstrated transmission of BASE to an alternative line of mice expressing wild-type levels of human PrP. In contrast, Beringue et al.113 showed transmission of BASE to mice overexpressing human PrP but no evidence of H-type BSE transmission, furthermore no evidence of CWD transmission to overexpressing mice has been identified.114,115 This difference in transmission results may be due to different genetic backgrounds or differences in PrP expression levels between the different mouse lines. An alternative to modeling the species barrier is the cell-free conversion assay which points to CWD as the animal prion disease with the greatest zoonotic potential, after (and very much less than) BSE.116 Surveillance Continued surveillance for long-term effects of BSE exposure in the UK human population appears necessary for the foreseeable future in order to discount possible second wave epidemics that might depend on genetic susceptibility, subclinical infection, and secondary transmission or disease in defined “at risk” patient groups such as hemophiliacs or patient groups in which full ascertainment is difficult, such as the elderly.
However, an additional concern is associated with idiopathic human prion disease. Sporadic CJD is not a uniform condition and the phenotype is clearly influenced by the codon 129 genotype of the patient and the prion protein type that accumulates in their brain. The etiological basis of the condition might be presumed to be spontaneously occurring, but this is not known with certainty in general, or in individual specific cases. Neither are the molecular mechanisms of spontaneous conversion of the prion protein to its pathogenic form well understood or easily investigated. Additionally, surveillance identifies apparently sporadic cases of human prion disease that do not fit well into currently accepted classification systems. This is exemplified by the recent identification of a new human prion disease (VPSPr by Gambetti et al.117) and its prospective and retrospective identification in other countries subsequently.118,110 The true prevalence, the relationship to sCJD and the risk to public health of VPSPr are yet to be determined.
Conclusions Since the identification of vCJD we have made progress in identifying routes of infection, controlling further infection, producing models of disease, developing decontamination procedures, and understanding susceptibility to disease. The vCJD epidemic in the UK now appears to be in decline and it appears that the control measures in food production and blood supplies have prevented further vCJD cases arising through dietary/ infected blood exposure. Despite this, there are still ongoing concerns over cases of vCJD arising in countries where little or no exposure to UK meat products have occurred, the presence of subclinical vCJD in the UK population with the possibility of further human-tohuman transmission and the identification of new strains of human prion disease. These scenarios necessitate ongoing studies in understanding transmission properties, disease diagnosis, and therapeutics. The identification of novel human prion diseases and the current estimates of subclinical vCJD infections show the importance of continued CJD surveillance and maintaining control measures already in place to protect human health.
2. Variably Protease-Sensitive Prionopathy
Professor James W Ironside and Dr Mark W Head, The National CJD Research and Surveillance Unit, University of Edinburgh.
Variably protease-sensitive prionopathy (VPSPr) is the most recently identified human prion disease, first described in the USA by Gambetti et al. in 2008 as “a novel human disease with abnormal prion protein sensitive to protease” [1]. Since then, similar cases have been identified in other countries; the National CJD Research and Surveillance Unit has identified nine cases in the UK, three of which have been identified retrospectively and the others prospectively from samples and data collected since 1991 [2-6]. Other candidate cases are currently under investigation.
Patients with VPSPr have no identified risk factors for acquired human prion disease and no associated mutations in the prion protein gene (PRNP) coding sequence have been found. In the original description a proportion of the patients had family histories of ill-defined dementia, but this has not been a feature in more recently identified cases [1,2,6]. VPSPr affects patients in the same age range as sporadic Creutzfeldt-Jakob disease (sCJD), occurring mostly in patients over the age of 60. The clinical features are more varied than in sCJD and include movement abnormalities, cognitive decline and unsteadiness while walking. The clinical illness is longer than for sCJD; most patients survive for over a year before succumbing to the illness. Diagnostic clinical criteria are therefore difficult to establish, and further work is required on this topic since this disease is likely to be under-ascertained [2,6].
Like sCJD, VPSPr occurs in all genetic groups defined by the polymorphism at codon 129 in the PRNP gene, ie MM, MV and MV. Unlike sCJD, there is a preponderance of the codon 129-V haplotype. VPSPr has distinctive neuropathological features, the most typical of which are microplaques that occur in a target-like arrangement and are particularly common in the cerebellum. These microplaques show differential staining with a panel of different anti-PrP antibodies, allowing a distinction from both the common sCJD VV2 and the rare sCJD VV1 subtypes [1,2,5,6]. The most distinctive and defining feature of VPSPr is the biochemistry of the abnormal prion protein in the brain, which is only poorly resistant to proteolytic digestion, yielding a low abundance, truncated 8kDa (approx) band in Western blot assays [1]. This fragment is often accompanied by a faint ladder of bands extending into the 18-30kDa range [1,2]. Some cases of VPSPr also show a sCJD-like pattern on Western blot analysis for abnormal prion protein, often in the cerebellum, suggesting molecular overlaps between VPSPr and sCJD [6,7].
Further work is required to fully establish the epidemiology, clinical and pathological diagnostic criteria and transmission characteristics of VPSPr. The Advisory Committee on Dangerous Pathogens Transmissible Spongiform Encephalopathy (ACDP TSE) Subgroup concluded that until further research can demonstrate how transmissible VPSPr may be, it would be advisable to add this novel form of human prion disease to the infection control guidance for CJD and other related disorders.
Subject: Variably protease-sensitive prionopathy (VPSPr)
USA CJD TSE Tables of Cases Examined National Prion Disease Pathology Surveillance Center Cases Examined May 1, 2018
http://prionunitusaupdate.blogspot.com/2018/07/usa-cjd-tse-tables-of-cases-examined.html
*** Conclusions. Preliminary results from transmission studies in bank voles strongly support the notion that VPSPr is a transmissible prion disease. Interestingly, VPSPr undergoes divergent evolution in the two genetic lines of voles, with sCJD-like features in BvM109 and GSS-like properties in BvI109. The discovery of previously unrecognized prion diseases in both humans and animals (i.e., Nor98 in small ruminants) demonstrates that the range of prion diseases might be wider than expected and raises crucial questions about the epidemiology and strain properties of these new forms. We are investigating this latter issue by molecular and biological comparison of VPSPr, GSS and Nor98.
OR-10: Variably protease-sensitive prionopathy is transmissible in bank voles
Romolo Nonno,1 Michele Di Bari,1 Laura Pirisinu,1 Claudia D’Agostino,1 Stefano Marcon,1 Geraldina Riccardi,1 Gabriele Vaccari,1 Piero Parchi,2 Wenquan Zou,3 Pierluigi Gambetti,3 Umberto Agrimi1 1 Istituto Superiore di Sanità; Rome, Italy; 2 Dipartimento di Scienze Neurologiche, Università di Bologna; Bologna, Italy; 3 Case Western Reserve University; Cleveland, OH USA
Background. Variably protease-sensitive prionopathy (VPSPr) is a recently described “sporadic”neurodegenerative disease involving prion protein aggregation, which has clinical similarities with non-Alzheimer dementias, such as fronto-temporal dementia. Currently, 30 cases of VPSPr have been reported in Europe and USA, of which 19 cases were homozygous for valine at codon 129 of the prion protein (VV), 8 were MV and 3 were MM. A distinctive feature of VPSPr is the electrophoretic pattern of PrPSc after digestion with proteinase K (PK). After PK-treatment, PrP from VPSPr forms a ladder-like electrophoretic pattern similar to that described in GSS cases. The clinical and pathological features of VPSPr raised the question of the correct classification of VPSPr among prion diseases or other forms of neurodegenerative disorders. Here we report preliminary data on the transmissibility and pathological features of VPSPr cases in bank voles.
Materials and Methods. Seven VPSPr cases were inoculated in two genetic lines of bank voles, carrying either methionine or isoleucine at codon 109 of the prion protein (named BvM109 and BvI109, respectively). Among the VPSPr cases selected, 2 were VV at PrP codon 129, 3 were MV and 2 were MM. Clinical diagnosis in voles was confirmed by brain pathological assessment and western blot for PK-resistant PrPSc (PrPres) with mAbs SAF32, SAF84, 12B2 and 9A2.
Results. To date, 2 VPSPr cases (1 MV and 1 MM) gave positive transmission in BvM109. Overall, 3 voles were positive with survival time between 290 and 588 d post inoculation (d.p.i.). All positive voles accumulated PrPres in the form of the typical PrP27–30, which was indistinguishable to that previously observed in BvM109 inoculated with sCJDMM1 cases. In BvI109, 3 VPSPr cases (2 VV and 1 MM) showed positive transmission until now. Overall, 5 voles were positive with survival time between 281 and 596 d.p.i.. In contrast to what observed in BvM109, all BvI109 showed a GSS-like PrPSc electrophoretic pattern, characterized by low molecular weight PrPres. These PrPres fragments were positive with mAb 9A2 and 12B2, while being negative with SAF32 and SAF84, suggesting that they are cleaved at both the C-terminus and the N-terminus. Second passages are in progress from these first successful transmissions.
Conclusions. Preliminary results from transmission studies in bank voles strongly support the notion that VPSPr is a transmissible prion disease. Interestingly, VPSPr undergoes divergent evolution in the two genetic lines of voles, with sCJD-like features in BvM109 and GSS-like properties in BvI109. The discovery of previously unrecognized prion diseases in both humans and animals (i.e., Nor98 in small ruminants) demonstrates that the range of prion diseases might be wider than expected and raises crucial questions about the epidemiology and strain properties of these new forms. We are investigating this latter issue by molecular and biological comparison of VPSPr, GSS and Nor98.
***Conclusions. Preliminary results from transmission studies in bank voles strongly support the notion that VPSPr is a transmissible prion disease. Interestingly, VPSPr undergoes divergent evolution in the two genetic lines of voles, with sCJD-like features in BvM109 and GSS-like properties in BvI109. The discovery of previously unrecognized prion diseases in both humans and animals (i.e., Nor98 in small ruminants) demonstrates that the range of prion diseases might be wider than expected and raises crucial questions about the epidemiology and strain properties of these new forms. We are investigating this latter issue by molecular and biological comparison of VPSPr, GSS and Nor98.
Variably protease-sensitive prionopathy in the UK: a retrospective review 1991–2008
Mark W. Head Helen M. Yull Diane L. Ritchie Jan P. Langeveld Nicholas A. Fletcher Richard S. Knight James W. Ironside Brain, Volume 136, Issue 4, 1 April 2013, Pages 1102–1115, https://doi.org/10.1093/brain/aws366 Published: 01 April 2013
Abstract
Variably protease-sensitive prionopathy is a newly described human prion disease of unknown aetiology lying out with the hitherto recognized phenotypic spectrum of Creutzfeldt–Jakob disease. Two cases that conform to the variably protease-sensitive prionopathy phenotype have been identified prospectively in the UK since the first description of the condition in 2008 in the USA. To determine the incidence and phenotype of variably protease-sensitive prionopathy within a single well-defined cohort, we have conducted a retrospective review of patients referred to the National Creutzfeldt–Jakob Disease Research & Surveillance Unit during the period 1991–2008. The approach taken was to screen frozen brain tissue by western blotting for the form of protease-resistant prion protein that characterizes variably protease-sensitive prionopathy, followed by neuropathological and clinical review of candidate cases. Cases diagnosed as sporadic Creutzfeldt–Jakob disease with atypical neuropathology were also reviewed. Four hundred and sixty-five cases were screened biochemically, yielding four candidate cases of variably protease-sensitive prionopathy. One was discounted on pathological and clinical grounds, and one was a known case of variably protease-sensitive prionopathy previously reported, leaving two new cases, which were confirmed biochemically and neuropathologically as variably protease-sensitive prionopathy. A third new case that lacked frozen tissue was recognized retrospectively on neuropathological grounds alone. This means that five cases of variably protease-sensitive prionopathy have been identified (prospectively and retrospectively) during the surveillance period 1991–2011 in the UK. Assuming ascertainment levels equivalent to that of other human prion diseases, these data indicate that variably protease-sensitive prionopathy is a rare phenotype within human prion diseases, which are themselves rare. Biochemical investigation indicates that the abnormal protease-resistant prion protein fragment that characterizes variably protease-sensitive prionopathy is detectable at low levels in some cases of sporadic Creutzfeldt–Jakob disease and conversely, that the form of abnormal prion protein that characterizes sporadic Creutzfeldt–Jakob disease can be found in certain brain regions of cases of variably protease-sensitive prionopathy, indicating molecular overlaps between these two disorders.
Creutzfeldt–Jakob disease, prion disease, variably protease-sensitive prionopathy, disease phenotype
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Variably protease-sensitive prionopathy is therefore transmissible, and in principle, it is possible that variably protease-sensitive prionopathy itself is acquired (Gambetti et al., 2011b; Nonno et al., 2012). The cases of variably protease-sensitive prionopathy that we have identified all occurred after the appearance of variant Creutzfeldt–Jakob disease in the UK in 1994–95 (Will et al., 1996), with deaths occurring in 1997 (VV), 2004 (VV), 2006 (MM), 2008 (VV) and 2008 (MV). The codon 129 distribution indicating PRNP codon 129V as a risk factor could be used to suggest that variably protease-sensitive prionopathy might represent bovine spongiform encephalopathy in non-129MM individuals; however, the reports of variably protease-sensitive prionopathy in other European countries and particularly in the USA (Gambetti et al., 2008) argue against variably protease-sensitive prionopathy being a bovine spongiform encephalopathy-related condition. Similarities in the protease-resistant prion protein type in variably protease-sensitive prionopathy and atypical scrapie could be used to argue for a link between these diseases, but there is no epidemiological evidence currently to support such an assertion. The structural similarity may be a sign of an analogous molecular aetiology, rather than evidence of a causative relationship. Evidence of a family history of dementia in the original cohort of cases with variably protease-sensitive prionopathy suggested a genetic basis, but no mutations in the PRNPopen reading frame have been found in our or previously reported cases. Therefore, a genetic basis would need to involve non-transcribed sequence of the PRNP gene or mutation of other loci (Gambetti et al., 2011b). At this point in time, the available evidence suggests that variably protease-sensitive prionopathy sits best within the spectrum of human prion diseases as an idiopathic form, alongside sporadic Creutzfeldt–Jakob disease and its subtypes, with which it shares clinical, pathological and, as we show here, biochemical features. Continued surveillance is required to help establish more accurately the prevalence and phenotypic range of this rare novel disorder.
***At this point in time, the available evidence suggests that variably protease-sensitive prionopathy sits best within the spectrum of human prion diseases as an idiopathic form, alongside sporadic Creutzfeldt–Jakob disease and its subtypes, with which it shares clinical, pathological and, as we show here, biochemical features.
BMJ Case Reports 2017; doi:10.1136/bcr-2017-220907
Rare disease
CASE REPORT
Gerstmann-Sträussler-Scheinker disease with atypical presentation
Sarah E Keuss1, James W Ironside2, Jonathan O’Riordan1
+ Author Affiliations
1Department of Neurology, Ninewells Hospital, Dundee, Tayside, UK
2Department of Clinical Brain Sciences, National Creutzfeldt-Jakob Disease Research and Surveillance Unit, Edinburgh, UK
Correspondence to Dr Jonathan O’Riordan, joriordan@nhs.net
Accepted 6 September 2017
Published 1 November 2017
Summary
We describe a 37-year-old woman who presented with progressive deafness, visual loss and ataxia. She latterly developed neuropsychiatric problems, including cognitive impairment, paranoid delusions and episodes of altered consciousness. She was found to be heterozygous for the Q212P mutation in the prion protein gene. She died over a decade after initial presentation and a diagnosis of prion disease was confirmed at postmortem.
P177 Transmission study of human prion diseases in PrP glycan-KO transgenic mice
Phd Silvio Notari1, PhD Laura Cracco1, PhD Satish Kumar Nemani1, MD Bernardino Ghetti2, PhD Qingzhong Kong1, MD Pierluigi Gambetti1
1Department of Pathology, Case Western Reserve University, Cleveland, United States, 2Department of Pathology and Laboratory Medicine, Indiana University, Indianapolis, United States
Aims:
The role of PrP glycosylation in various transmission barriers including species, mutation and 129 polymorphism barriers has been studied in several in vitro and in vivo models. We report an update of our studies on the propensity to transmit and reproduce the histopathological phenotype human prion diseases, including familial and sporadic forms historically difficult to transmit to humanized transgenic mice [Tg(HuPrP)]. Hamster Hyperactive (Hyper) and Drowsy prion strains have also been examined.
Methods:
We generated a new transgenic mouse line expressing unglycosylated PrP-129M [Tg(HuPrP-129MGlycKO)], here after referred to as TgNN6h, by substituting asparagine with glutamine at both residue 181 and 197 glycosylation sites (N181Q/N197Q). TgNN6h were inoculated with different human prion strains: Variably protease sensitive prionopathy (VPSPr), Gerstmann-Sträussler-Scheinker (GSS) mutations (A117V, P102L8kDa, P102L21kDa, F198S), sporadic Creutzfeldt-Jakob disease (sCJD, subtypes MM2, VV2, MM1, VV1, MV2K, MV2C), familial CJD V180I (fCJDv180I), sporadic fatal insomnia (sFI) as well as hamster Hyper and Drowsy strains.
Results:
TgNN6h efficiently transmitted VPSPr with attack rates (ARs) and incubation periods (IPs) changed as a function of the VPSPr 129 genotype (highest in 129MM/MV and lowest in 129VV). To note that the transmission to TgNN6h was relatively efficient even with VPSPr-VV despite the 129 genotypic barrier, which is totally divergent from the lack of transmission of VPSPr-VV to Tg mice expressing wild type human PrP [Tg(HuWTPrP)].
Among GSS mutations subtypes, P102L21kDa, F198S, P102L8kDa and A117V were transmitted with various efficiencies. Despite limited neuropathology and medium-long average IP (380-700 days), a high rate of GSS-inoculated animals suffered of typical prion-related clinical signs before death, a finding hinting to the presence of PrPSc (investigation in progress).
Familial CJD V180I was easily transmitted with AR, IP and phenotype comparable to those of VPSPr, confirming the conformation similarity of the PrPSc species in these two diseases.
Within the sCJD group, sCJDMM2 and sFI were the only conditions that transmitted to TgNN6h much more efficiently than to [Tg(HuWTPrP)] with IP reductions of approximately 50% and 90% at first and second passage, respectively.
Hamster Hyper and Drowsy strains did not transmit efficiently to TgNN6h.
Conclusions:
Our data indicate that glycan-free TgNN6h may play a role in testing experimental transmission and studying transmission barriers of human prion diseases, especially those difficult to transmit to Tg mice expressing wild type human PrPC.
Compared to previous studies, our data underscore the variability of the response to transmission between different models of glycan-free Tg mice.
DISORDERS PRION 2017 DECIPHERING NEURODEGENERATIVE
DISORDERS PRION 2017 DECIPHERING NEURODEGENERATIVE
Gerstmann-Sträussler-Scheinker disease subtypes efficiently transmit in bank voles as genuine prion diseases
Published online: 04 February 2016
Tuesday, November 29, 2016
Transmissibility of Gerstmann–Sträussler–Scheinker syndrome in rodent models: new insights into the molecular underpinnings of prion infectivity
2015 PRION CONFERENCE
*** RE-P.164: Blood transmission of prion infectivity in the squirrel monkey: The Baxter study
***suggest that blood donations from cases of GSS (and perhaps other familial forms of TSE) carry more risk than from vCJD cases, and that little or no risk is associated with sCJD. ***
P.164: Blood transmission of prion infectivity in the squirrel monkey: The Baxter study
Paul Brown1, Diane Ritchie2, James Ironside2, Christian Abee3, Thomas Kreil4, and Susan Gibson5 1NIH (retired); Bethesda, MD USA; 2University of Edinburgh; Edinburgh, UK; 3University of Texas; Bastrop, TX USA; 4Baxter Bioscience; Vienna, Austria; 5University of South Alabama; Mobile, AL USA
Five vCJD disease transmissions and an estimated 1 in 2000 ‘silent’ infections in UK residents emphasize the continued need for information about disease risk in humans. A large study of blood component infectivity in a non-human primate model has now been completed and analyzed. Among 1 GSS, 4 sCJD, and 3 vCJD cases, only GSS leukocytes transmitted disease within a 5–6 year surveillance period. A transmission study in recipients of multiple whole blood transfusions during the incubation and clinical stages of sCJD and vCJD in ic-infected donor animals was uniformly negative. These results, together with other laboratory studies in rodents and nonhuman primates and epidemiological observations in humans, suggest that blood donations from cases of GSS (and perhaps other familial forms of TSE) carry more risk than from vCJD cases, and that little or no risk is associated with sCJD. The issue of decades-long incubation periods in ‘silent’ vCJD carriers remains open.
ran across an old paper from 1984 ;
***The occurrence of contact cases raises the possibility that transmission in families may be effected by an unusually virulent strain of the agent. ***
EVIDENCE FOR CASE-TO-CASE TRANSMISSION OF C.J.D.
The possible iatrogenic transmission of C.J.D. by neurosurgery, corneal transplantation and stereotactic electrodes has been suggested in the past (Duffy et al., 1974: Bernouilli et al., 1977; Masters et al., 1979a). In this series the close temporal relationship of neurosurgical procedures on two affected patients and three patients, unaffected at the time but who subsequently developed the disease is described. This provides strong circumstantial evidence of iatrogenic transmission by neurosurgery. Although sterilisation procedures have improved since the cases described, the unusual resistance of the agent and the recent description of probable neurosurgical transmission in France (Foncin et al., 1980) suggests that there is a continued risk of accidental transmission. However, brain biopsy to confirm the diagnosis of C.J.D. is now an unusual event and computed tomography has obviated the need for ventriculography.
The depth electrodes putatively responsible for one case of iatrogenic transmission in this series were inadequately sterilised in formalin and were subsequently used in over 200 patients. The neurosurgical instruments used in the cases of presumed neurosurgical transmission were sterilised using autoclaving procedures which were inadequate according to current advice (Gajdusek et al., 1978). However, despite detailed investigation, no cases other than those described above are known to have developed C.J.D. Thus, despite the possible implantation of the agent directly into the central nervous system, a large number of patients failed to develop the disease. This provides circumstantial evidence of an inherited susceptibility to the agent and suggests that cases of iatrogenic transmission may have occurred due to the unfortunate temporal proximity of susceptible individuals exposed to the agent.
In the close geographic group of three cases possible nodes of transmission can be suggested, either iatrogenic or through dental procedures, but these must remain conjectural. It is known, however, that the similar scrapie agent can be transmitted from the gums of animals (Adams and Edgar, 1978). Such close spatial clustering of cases is extremely unusual, being previously reported in England (Matthews, 1975a), Czechoslovakia (Mayer et al., 1977) and Hungary (Majtenyi, 1978), but not detected in the study of the epidemiology of C.J.D. in urban Paris (Cathala et al., 1978) where the incidence was found to be relatively high.
The occurrence of the disease in a patient who had contact with cases of familial C.J.D., but was not genetically related, has been described in Chile (Galvez et al., 1980) and in France (Brown et al., 1979b). In Chile the patient was related by marriage, but with no consanguinity, and had social contact with subsequently affected family members for 13 years before developing the disease. The contact case in France also married into a family in which C.J.D. was prevalent and had close contact with an affected member. In neither instance did the spouse of the non-familial case have the disease. The case described in this report was similarly related to affected family members and social contact had occurred for 20 years prior to developing C.J.D. If contact transmission had occurred, the minimum transmission period would be 11 years. Contact between sporadic cases has not been described and it is remarkable that possible contact transmissions have all been with familial cases. No method of transmission by casual social contact has been suggested.
***The occurrence of contact cases raises the possibility that transmission in families may be effected by an unusually virulent strain of the agent.
snip...see full text here;
snip...see full text ;
>>>Although sCJD patient skin contained ~103- to 105-fold lower prion seeding activity than did sCJD patient brain tissue, all 12 mice from two transgenic mouse lines inoculated with sCJD skin homogenates from two sCJD patients succumbed to prion disease within 564 days after inoculation<<<
for something to be 103 to 105 fold lower prion seeding, yet still be 100% fatal in all test subjects, very disturbing...terry
WEDNESDAY, NOVEMBER 22, 2017
NIH scientists and collaborators find infectious prion protein in skin of CJD patients
TUESDAY, JUNE 20, 2017
Prion 2017 Conference
Transmissible prions in the skin of Creutzfeldt-Jakob disease patients
Prion 2017 Conference Transmissible prions in the skin of Creutzfeldt-Jakob disease patients
Dr. Wenguan Zou1, Dr. Christina Orru2, Jue Yuan1, Brian Appleby1, Baiya Li1, Dane Winner1, Yian Zhan1,3, Mark Rodgers1, Jason Rarick1, Robert Wyza1, Tripti Joshi1, Gongxian Wang3, Mark Cohen1, Shulin Zhang1, Bradley Groveman2, Robert Petersen1, James Ironside4, Miguel Quinones-Mateu1, Jiri Safar1, Qingzhong Kong1, Byron Caughey2
1Case Western Reserve University, Cleveland, United States, 2Rocky Mountain Laboratories, National Institutes of Health, Hamilton, United States, 3Nanchang University, Nanchang, China, 4Universitv of Edinburgh, Edinburgh, United Kingdom
Aims: Sporadic Creutzfeldt-Jakob disease (sCJD), the most common human prion disease, is transmissible by neuroinvasive iatrogenic routes due to abundant prion infectivity in the central nervous system (CNS). The disease-associated prion protein (PrPSc) and its infectivity have never been detected in skin from sCJD patients; however, some epidemiological studies have associated sCJD risk with skin-involved non-CNS surgeries. The aims of our study were to explore potential prion seeding activity and infectivity of skin and the feasibility of skin-based CJD diagnosis.
Methods: Skin samples were collected at autopsy or biopsy from twenty-one sCJD, two variant CJD, and fifteen non-CJD patients and analysed by Western blotting and real-time quaking-induced conversion (RT- QulC) for PrPSc. Infectivity of skin from two sCJD patients was determined by bioassay using two lines of humanized transgenic (Tg) mice.
Results: Western blotting demonstrated PrPSc in the skin of one of five deceased sCJD patients examined. However, the more sensitive RT-QuIC assay detected prion-seeding activity in skin from all 23 CJD decedents but not in non-CJD controls, indicating preliminary ClD diagnostic sensitivities and specificities of 100% (95% confidence intervals of 85-100%, and 78-100%, respectively). Although sCJD skins contained ~102-105-fold lower RT-QuIC seeding activity than sCJD brains, ten out of twelve mice from two Tg mouse lines inoculated with skin homogenates of two patients with two different subtypes of sCJD succumbed to prion disease within 450 days after inoculation.
Conclusions: sCJD patients' skin may contain both detectable prion seeding activity and transmissible prions. Our findings not only suggest a new basis for diagnostic sCJD testing, but also raise concerns about the potential for iatrogenic sCJD transmission via skin. (Funded by the CJD Foundation, the National Institute of Neurological Disorders and Stroke, the Centers for Disease Control and Prevention, as well as others)
DISORDERS PRION 2017 DECIPHERING NEURODEGENERATIVE
*sCJD patients' skin may contain both detectable prion seeding activity and transmissible prions.
*Our findings not only suggest a new basis for diagnostic sCJD testing, but also raise concerns about the potential for iatrogenic sCJD transmission via skin.
Oral Session14：45～15：00O-12 Wenquan Zou
*** PrPSc in the skin of CJD patients
Accessing transmissibility and diagnostic marker of skin prions.
Kong, Qingzhong Safar, Jiri G. Zou, Wen-Quan
Case Western Reserve University, Cleveland, OH, United States
Abstract The fatal, transmissible animal and human prion diseases are characterized by the deposition in the brain of a proteinase K (PK)-resistant infectious prion protein (PrPSc), an isoform derived from the cellular protein (PrPC) through misfolding. A definitive antemortem diagnosis is virtually impossible for most patients because of the difficulty in obtaining the brain tissues by biopsy. Recently, PrPSc has been reported to be detected in the skin of experimentally or naturally scrapie-infected animals (Thomzig et al., 2007). Consistent with this finding, we have observed PK-resistant PrP in the skin of a patient with variant Creutzfeldt-Jakob disease (vCJD), an acquired form of human prion disease caused by bovine prion (Notari et al., 2010). Unexpectedly, our latest preliminary study identified two types of PK-resistant PrP molecules [with gel mobility similar to the PrPSc types 1 and 2 from the brain of sporadic CJD (sCJD)] in the fibroblast cells extracted from the skin of clinical sCJD patients and asymptomatic subjects carrying PrP mutations linked to familial CJD (fCJD). We also detected PrPSc in the skin of humanized transgenic (Tg) mice inoculated intracerebrally with a human prion. Moreover, prion infectivity has been observed in the skin of infected greater kudu (Cunningham et al., 2004) and a murine prion inoculated to mice via skin scarification can not only propagate in the skin, but also spread to the brain to cause prion disease (Wathne et al., 2012). We hypothesize that the skin of patients with prion disease harbors prion infectivity and the presence of PK-resistant PrP in the skin is a novel diagnostic marker for preclinical CJD patients. To test the hypotheses, we propose to (1) determine prion infectivity of the skin- derived fibroblasts and skin of sCJD patients and asymptomatic PrP-mutation carriers using humanized Tg mouse bioassay, (2) to pinpoint the earliest stage at which PrPSc becomes detectable in the skin of prion- infected Tg mice, and (3) to detect PrPSc in the skin of various human prion diseases, using conventional as well as highly sensitive RT-QuIC assays for both (2) and (3). If successful, our proposal may not only help prevent potential transmission of human prion diseases but also enable definitive and less intrusive antemortem diagnosis of prion diseases. Finally, knowledge generated from this study may also enhance our understanding of other neurodegenerative diseases such as Alzheimer's disease.
Public Health Relevance Currently it is unclear whether or not the skin of patients with prion diseases is infectious and, moreover, there is no alternative preclinical definitive testing or the brain biopsy in the prion diseases. The aim of our proposal is to address the issues by detection of the infectivity of patients' skin samples using animal bioassay and a new highly sensitive RT-QuIC assay. We believe that our study will not only provide insights into the pathogenesis and transmissibility of prion disease but also will develop preclinical definitive testing for prion disease.
Funding Agency Agency National Institute of Health (NIH)
Institute National Institute of Neurological Disorders and Stroke (NINDS)
Type Exploratory/Developmental Grants (R21)
Project # 1R21NS096626-01
Application # 9092119
Study Section Special Emphasis Panel (ZRG1)
Program Officer Wong, May Project Start 2016-02-01
Project End 2018-01-31
Budget Start 2016-02-01
Budget End 2017-01-31
Support Year 1
Fiscal Year 2016
Total Cost
Indirect Cost Institution Name Case Western Reserve University
Department Pathology
Type Schools of Medicine
DUNS # 077758407
City Cleveland
State OH
Country United States
Zip Code 44106
TUESDAY, MAY 10, 2016
Accessing transmissibility and diagnostic marker of skin prions
Variably protease-sensitive prionopathy (VPSPr), a recently identified and seemingly sporadic human prion disease, is distinct from Creutzfeldt-Jakob disease (CJD) but shares features of Gerstmann-Sträussler-Scheinker disease (GSS). However, contrary to exclusively inherited GSS, no prion protein (PrP) gene variations have been detected in VPSPr, suggesting that VPSPr might be the long-sought sporadic form of GSS. snip...
In conclusion, we propose that VPSPr is transmissible and, therefore, is an authentic prion disease. However, transmissibility cannot be sustained through serial passages presumably because human PrPC (or the mouse brain environment) cannot efficiently convert and propagate the VPSPr PrPSc species. If this is the case, uncovering the properties of human PrP that are required to replicate more efficiently the prion strains associated with VPSPr may help clarify the PrPSc mode of formation in this intriguing disease.
WEDNESDAY, NOVEMBER 09, 2011
Case report Sporadic fatal insomnia in a young woman: A diagnostic challenge: Case Report TEXAS
HOW TO TURN A POTENTIAL MAD COW VICTIM IN THE USA, INTO A HAPPENSTANCE OF BAD LUCK, A SPONTANEOUS MUTATION FROM NOTHING.
OR WAS IT $$$
Confirmed Variant Creutzfeldt-Jakob Disease (variant CJD) Case in Texas
Updated: October 7, 2014
CDC and the Texas Department of State Health Services (DSHS) have completed the investigation of the recently reported fourth vCJD case in the United States. It confirmed that the case was in a US citizen born outside the Americas and indicated that the patient's exposure to the BSE/vCJD agent most likely occurred before he moved to the United States; the patient had resided in Kuwait, Russia and Lebanon. The completed investigation did not support the patient's having had extended travel to European countries, including the United Kingdom, or travel to Saudi Arabia. The specific overseas country where this patient’s infection occurred is less clear largely because the investigation did not definitely link him to a country where other known vCJD cases likely had been infected.
FRIDAY, NOVEMBER 3, 2017
GSS Gerstmann-Sträussler-Scheinker disease with atypical presentation
Gerstmann-Sträussler-Scheinker disease subtypes efficiently transmit in bank voles as genuine prion diseases
Laura Pirisinu, Michele A. Di Bari, Claudia D’Agostino, Stefano Marcon, Geraldina Riccardi, Anna Poleggi, Mark L. Cohen, Brian S. Appleby, Pierluigi Gambetti, Bernardino Ghetti, Umberto Agrimi & Romolo Nonno
Abstract
Gerstmann-Sträussler-Scheinker disease (GSS) is an inherited neurodegenerative disorder associated with mutations in the prion protein gene and accumulation of misfolded PrP with protease-resistant fragments (PrPres) of 6–8 kDa. With the exception of a few GSS cases characterized by co-accumulation of PrPres of 21 kDa, efforts to transmit GSS to rodents have been unsuccessful. As a result, GSS subtypes exclusively associated with 6–8 kDa PrPres have often been considered as non-transmissible proteinopathies rather than true prion diseases. We show that GSS with P102L, A117V and F198S mutations transmit efficiently and produce distinct pathological phenotypes in bank voles (M. glareolus), irrespective of the presence of 21 kDa PrPres in the inoculum, demonstrating that GSS is a genuine prion disease characterized by both transmissibility and strain variation.
snip...
Inefficient transmission of GSS cases with 6–8 kDa PrPres has led to the hypothesis that GSS is a non-transmissible proteinopathy rather than authentic prion disease5, implying that proteinopathies and prion diseases were associated with different isoforms of PrPSc. Indeed, recent studies suggest differences between cell-to-cell spread of misfolded proteins and the transfer of infectivity from one organism to another23, as well as between pathogenic and infectious properties of the prion protein itself 24. To address these issues, we attempted to transmit several biochemically characterized GSS cases10 to bank voles, a rodent model able to propagate most human and animal prion diseases25,26,27,28,29,30,31,32,33. We found that all GSS cases induced prion diseases characterized by spongiform degeneration, PrPSc deposition, and propagation of infectious agents in the brain of recipient voles, implying that GSS, similar to others TSEs, is associated with infectious prions.
snip...
We inoculated seven distinct GSS cases into Bv109I and observed disease transmission in all cases. Overall, 85/113 voles challenged with brain homogenates from different GSS patients developed a fatal neurological illness, most of them between 3 and 7 months post-challenge. Infected voles showed all cardinal neuropathological and molecular features of prion diseases, including spongiform degeneration and deposition of PK-resistant PrPSc. Furthermore, GSS A117V infected vole brains were able to induce the same disease phenotype in recipient voles within 3–4 months after challenge, proving that a prion agent propagated in the brains of infected animals. These findings imply that brains of GSS patients harbor infectious prions with transmissibility features similar to those found in other human and animal TSEs.
snip...see full text;
Extra Views
Transmissibility of Gerstmann–Sträussler–Scheinker syndrome in rodent models: New insights into the molecular underpinnings of prion infectivity
Romolo Nonno, Michele Angelo Di Bari, Umberto Agrimi & Laura Pirisinu
Pages 421-433 | Received 10 Aug 2016, Accepted 19 Sep 2016, Accepted author version posted online: 28 Nov 2016, Published online: 28 Nov 2016 Download citation https://doi.org/10.1080/19336896.2016.1239686 ;
ABSTRACT
Prion diseases, or transmissible spongiform encephalopathies, have revealed the bewildering phenomenon of transmissibility in neurodegenerative diseases. Hence, the experimental transmissibility of prion-like neurodegenerative diseases via template directed misfolding has become the focus of intense research. Gerstmann-Sträussler-Scheinker disease (GSS) is an inherited prion disease associated with mutations in the prion protein gene. However, with the exception of a few GSS cases with P102L mutation characterized by co-accumulation of protease-resistant PrP core (PrPres) of ∼21 kDa, attempts to transmit to rodents GSS associated to atypical misfolded prion protein with ∼8 kDa PrPres have been unsuccessful. As a result, these GSS subtypes have often been considered as non-transmissible proteinopathies rather than true prion diseases. In a recent study we inoculated bank voles with GSS cases associated with P102L, A117V and F198S mutations and found that they transmitted efficiently and produced distinct pathological phenotypes, irrespective of the presence of 21 kDa PrPres in the inoculum. This study demonstrates that GSS is a genuine prion disease characterized by both transmissibility and strain variation. We discuss the implications of these findings for the understanding of the heterogeneous clinic-pathological phenotypes of GSS and of the molecular underpinnings of prion infectivity.
snip...
Taken together, transmission experiments suggest that P102L GSS cases with 21 kDa PrPres and possibly with extensive spongiform degeneration are associated with classical transmissibility of PrP misfolding and prion disease, while these studies fall short in providing definitive proof that GSS forms characterized by 8 kDa PrPres actually contain transmissible agents. In keeping with evidences from transgenic mouse models of spontaneous disease, this might lead to hypothesize that the atypical PrPSc conformers which characterize most GSS cases could be associated with neurotoxicity, but not with “true” infectivity. Some support to this hypothesis derives from the study of a newly described sporadic prion disease, Variably Protease Sensitive Prionopathy (VPSPr). VPSPr is characterized by an atypical PrPSc isoform with a ∼7 kDa PK-resistant core, similar to GSS, suggesting that VPSPr might be the long-sought sporadic form of GSS.32
Gambetti P, Dong Z, Yuan J, Xiao X, Zheng M, Alshekhlee A, Castellani R, Cohen M, Barria MA, Gonzalez-Romero D, et al. A novel human disease with abnormal prion protein sensitive to protease. Ann Neurol 2008; 63:697-708; PMID:18571782; http://dx.doi.org/10.1002/ana.21420
KEYWORDS: Gerstmann-Straussler-Scheinker disease, neurodegenerative diseases, prion infectivity, protein misfolding, strain, PrPC, PrPSc
snip...see full text;
A case of sporadic Creutzfeldt-Jakob disease with a Gerstmann-Sträussler-Scheinker phenotype but no alterations in the PRNP gene
P. P. Liberski · M. Barcikowska · L. Cervenakova · J. Bratosiewicz · M. Marczewska · P. Brown · D. C. Gajdusek
Acta Neuropathol (1998) 96 : 425-430 © Springer-Verlag 1998 Received: 15 July 1997 / Revised, accepted: 24 March 1998
Key words Prions · Creutzfeldt-Jakob disease · Gerstmann-Sträussler-Scheinker disease
Abstract
We report here an unusual sporadic case of Creutzfeldt-Jakob disease (CJD) characterized by an abundance of prion protein (PrP)-immunopositive kuru and multicentric but not florid plaques. Molecular genetic analysis of the PRNP open reading frame region spanning codons 8-221 was performed. Neither deletion nor insertion mutations were detected in the repeat area of the PRNP. No pathogenic mutation was found in the sequenced region between codon 108-221. Restriction analysis of the amplified fragment using restriction endonucleases DdeI, PvuII and AluI did not show any of the previously described pathogenic mutations at codon 102, 105, and 117 associated with Gerstmann-Sträussler-Scheinker (GSS). The patient was heterozygous for the methionine/ valine coding triplet at polymorphic codon 129 of the PRNP gene by sequence, restriction endonuclease analysis and hybridization with allele-specific nucleotides. Furthermore, hybridization with 32P-labeled allele-specific oligonucleotides confirmed the absence of pathogenic mutations at codons 102, 200 and 178. Such a case may present a missing "link" between sporadic CJD and familial GSS.
A case of sporadic Creutzfeldt-Jakob disease with a Gerstmann-Sträussler-Scheinker phenotype but no alterations in the PRNP gene
P. P. Liberski · M. Barcikowska · L. Cervenakova · J. Bratosiewicz · M. Marczewska · P. Brown · D. C. Gajdusek
Acta Neuropathol (1998) 96 : 425-430 © Springer-Verlag 1998 Received: 15 July 1997 / Revised, accepted: 24 March 1998
Key words Prions · Creutzfeldt-Jakob disease · Gerstmann-Sträussler-Scheinker disease
Abstract
We report here an unusual sporadic case of Creutzfeldt-Jakob disease (CJD) characterized by an abundance of prion protein (PrP)-immunopositive kuru and multicentric but not florid plaques. Molecular genetic analysis of the PRNP open reading frame region spanning codons 8-221 was performed. Neither deletion nor insertion mutations were detected in the repeat area of the PRNP. No pathogenic mutation was found in the sequenced region between codon 108-221. Restriction analysis of the amplified fragment using restriction endonucleases DdeI, PvuII and AluI did not show any of the previously described pathogenic mutations at codon 102, 105, and 117 associated with Gerstmann-Sträussler-Scheinker (GSS). The patient was heterozygous for the methionine/ valine coding triplet at polymorphic codon 129 of the PRNP gene by sequence, restriction endonuclease analysis and hybridization with allele-specific nucleotides. Furthermore, hybridization with 32P-labeled allele-specific oligonucleotides confirmed the absence of pathogenic mutations at codons 102, 200 and 178. Such a case may present a missing "link" between sporadic CJD and familial GSS.
Ann Neurol. 2010 Aug;68(2):162-72.
Variably protease-sensitive prionopathy: a new sporadic disease of the prion protein.
Zou WQ, Puoti G, Xiao X, Yuan J, Qing L, Cali I, Shimoji M, Langeveld JP, Castellani R, Notari S, Crain B, Schmidt RE, Geschwind M, Dearmond SJ, Cairns NJ, Dickson D, Honig L, Torres JM, Mastrianni J, Capellari S, Giaccone G, Belay ED, Schonberger LB, Cohen M, Perry G, Kong Q, Parchi P, Tagliavini F, Gambetti P.
Department of Pathology, National Prion Disease Pathology Surveillance Center, Case Western Reserve University, Cleveland, OH 44106, USA. wenquan.zou@case.edu
Abstract OBJECTIVE: The objective of the study is to report 2 new genotypic forms of protease-sensitive prionopathy (PSPr), a novel prion disease described in 2008, in 11 subjects all homozygous for valine at codon 129 of the prion protein (PrP) gene (129VV). The 2 new PSPr forms affect individuals who are either homozygous for methionine (129MM) or heterozygous for methionine/valine (129MV).
METHODS: Fifteen affected subjects with 129MM, 129MV, and 129VV underwent comparative evaluation at the National Prion Disease Pathology Surveillance Center for clinical, histopathologic, immunohistochemical, genotypical, and PrP characteristics.
RESULTS: Disease duration (between 22 and 45 months) was significantly different in the 129VV and 129MV subjects. Most other phenotypic features along with the PrP electrophoretic profile were similar but distinguishable in the 3 129 genotypes. A major difference laid in the sensitivity to protease digestion of the disease-associated PrP, which was high in 129VV but much lower, or altogether lacking, in 129MV and 129MM. This difference prompted the substitution of the original designation with "variably protease-sensitive prionopathy" (VPSPr). None of the subjects had mutations in the PrP gene coding region.
INTERPRETATION: Because all 3 129 genotypes are involved, and are associated with distinguishable phenotypes, VPSPr becomes the second sporadic prion protein disease with this feature after Creutzfeldt-Jakob disease, originally reported in 1920. However, the characteristics of the abnormal prion protein suggest that VPSPr is different from typical prion diseases, and perhaps more akin to subtypes of Gerstmann-Sträussler-Scheinker disease.
PMID: 20695009 [PubMed - indexed for MEDLINE]
None of the subjects had mutations in the PrP gene coding region....???...TSS
see much more here;
PR-26
NOR98 SHOWS MOLECULAR FEATURES REMINISCENT OF GSS
R. Nonno1, E. Esposito1, G. Vaccari1, E. Bandino2, M. Conte1, B. Chiappini1, S. Marcon1, M. Di Bari1, S.L. Benestad3, U. Agrimi1 1 Istituto Superiore di Sanità, Department of Food Safety and Veterinary Public Health, Rome, Italy (romolo.nonno@iss.it); 2 Istituto Zooprofilattico della Sardegna, Sassari, Italy; 3 National Veterinary Institute, Department of Pathology, Oslo, Norway
Molecular variants of PrPSc are being increasingly investigated in sheep scrapie and are generally referred to as “atypical” scrapie, as opposed to “classical scrapie”. Among the atypical group, Nor98 seems to be the best identified. We studied the molecular properties of Italian and Norwegian Nor98 samples by WB analysis of brain homogenates, either untreated, digested with different concentrations of proteinase K, or subjected to enzymatic deglycosylation. The identity of PrP fragments was inferred by means of antibodies spanning the full PrP sequence. We found that undigested brain homogenates contain a Nor98-specific PrP fragment migrating at 11 kDa (PrP11), truncated at both the C-terminus and the N-terminus, and not N-glycosylated. After mild PK digestion, Nor98 displayed full-length PrP (FL-PrP) and N-glycosylated C-terminal fragments (CTF), along with increased levels of PrP11. Proteinase K digestion curves (0,006-6,4 mg/ml) showed that FL-PrP and CTF are mainly digested above 0,01 mg/ml, while PrP11 is not entirely digested even at the highest concentrations, similarly to PrP27-30 associated with classical scrapie. Above 0,2 mg/ml PK, most Nor98 samples showed only PrP11 and a fragment of 17 kDa with the same properties of PrP11, that was tentatively identified as a dimer of PrP11. Detergent solubility studies showed that PrP11 is insoluble in 2% sodium laurylsorcosine and is mainly produced from detergentsoluble, full-length PrPSc. Furthermore, among Italian scrapie isolates, we found that a sample with molecular and pathological properties consistent with Nor98 showed plaque-like deposits of PrPSc in the thalamus when the brain was analysed by PrPSc immunohistochemistry. Taken together, our results show that the distinctive pathological feature of Nor98 is a PrP fragment spanning amino acids ~ 90-155. This fragment is produced by successive N-terminal and C-terminal cleavages from a full-length and largely detergent-soluble PrPSc, is produced in vivo and is extremely resistant to PK digestion. Intriguingly, these conclusions suggest that some pathological features of Nor98 are reminiscent of Gerstmann-Sträussler-Scheinker disease.
119
P03.141
Aspects of the Cerebellar Neuropathology in Nor98
Gavier-Widén, D1; Benestad, SL2; Ottander, L1; Westergren, E1 1National Veterinary Insitute, Sweden; 2National Veterinary Institute,
Norway Nor98 is a prion disease of old sheep and goats. This atypical form of scrapie was first described in Norway in 1998. Several features of Nor98 were shown to be different from classical scrapie including the distribution of disease associated prion protein (PrPd) accumulation in the brain. The cerebellum is generally the most affected brain area in Nor98. The study here presented aimed at adding information on the neuropathology in the cerebellum of Nor98 naturally affected sheep of various genotypes in Sweden and Norway. A panel of histochemical and immunohistochemical (IHC) stainings such as IHC for PrPd, synaptophysin, glial fibrillary acidic protein, amyloid, and cell markers for phagocytic cells were conducted. The type of histological lesions and tissue reactions were evaluated. The types of PrPd deposition were characterized. The cerebellar cortex was regularly affected, even though there was a variation in the severity of the lesions from case to case. Neuropil vacuolation was more marked in the molecular layer, but affected also the granular cell layer. There was a loss of granule cells. Punctate deposition of PrPd was characteristic. It was morphologically and in distribution identical with that of synaptophysin, suggesting that PrPd accumulates in the synaptic structures. PrPd was also observed in the granule cell layer and in the white matter. The pathology features of Nor98 in the cerebellum of the affected sheep showed similarities with those of sporadic Creutzfeldt-Jakob disease in humans.
A newly identified type of scrapie agent can naturally infect sheep with resistant PrP genotypes
Annick Le Dur*,?, Vincent Béringue*,?, Olivier Andréoletti?, Fabienne Reine*, Thanh Lan Laï*, Thierry Baron§, Bjørn Bratberg¶, Jean-Luc Vilotte?, Pierre Sarradin**, Sylvie L. Benestad¶, and Hubert Laude*,?? +Author Affiliations
*Virologie Immunologie Moléculaires and ?Génétique Biochimique et Cytogénétique, Institut National de la Recherche Agronomique, 78350 Jouy-en-Josas, France; ?Unité Mixte de Recherche, Institut National de la Recherche Agronomique-Ecole Nationale Vétérinaire de Toulouse, Interactions Hôte Agent Pathogène, 31066 Toulouse, France; §Agence Française de Sécurité Sanitaire des Aliments, Unité Agents Transmissibles Non Conventionnels, 69364 Lyon, France; **Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, 37380 Nouzilly, France; and ¶Department of Pathology, National Veterinary Institute, 0033 Oslo, Norway
Edited by Stanley B. Prusiner, University of California, San Francisco, CA (received for review March 21, 2005)
Abstract Scrapie in small ruminants belongs to transmissible spongiform encephalopathies (TSEs), or prion diseases, a family of fatal neurodegenerative disorders that affect humans and animals and can transmit within and between species by ingestion or inoculation. Conversion of the host-encoded prion protein (PrP), normal cellular PrP (PrPc), into a misfolded form, abnormal PrP (PrPSc), plays a key role in TSE transmission and pathogenesis. The intensified surveillance of scrapie in the European Union, together with the improvement of PrPSc detection techniques, has led to the discovery of a growing number of so-called atypical scrapie cases. These include clinical Nor98 cases first identified in Norwegian sheep on the basis of unusual pathological and PrPSc molecular features and "cases" that produced discordant responses in the rapid tests currently applied to the large-scale random screening of slaughtered or fallen animals. Worryingly, a substantial proportion of such cases involved sheep with PrP genotypes known until now to confer natural resistance to conventional scrapie. Here we report that both Nor98 and discordant cases, including three sheep homozygous for the resistant PrPARR allele (A136R154R171), efficiently transmitted the disease to transgenic mice expressing ovine PrP, and that they shared unique biological and biochemical features upon propagation in mice. These observations support the view that a truly infectious TSE agent, unrecognized until recently, infects sheep and goat flocks and may have important implications in terms of scrapie control and public health.
Monday, December 1, 2008
When Atypical Scrapie cross species barriers
Authors
Andreoletti O., Herva M. H., Cassard H., Espinosa J. C., Lacroux C., Simon S., Padilla D., Benestad S. L., Lantier F., Schelcher F., Grassi J., Torres, J. M., UMR INRA ENVT 1225, Ecole Nationale Veterinaire de Toulouse.France; ICISA-INlA, Madrid, Spain; CEA, IBiTec-5, DSV, CEA/Saclay, Gif sur Yvette cedex, France; National Veterinary Institute, Postboks 750 Sentrum, 0106 Oslo, Norway, INRA IASP, Centre INRA de Tours, 3738O Nouzilly, France.
Content
Atypical scrapie is a TSE occurring in small ruminants and harbouring peculiar clinical, epidemiological and biochemical properties. Currently this form of disease is identified in a large number of countries. In this study we report the transmission of an atypical scrapie isolate through different species barriers as modeled by transgenic mice (Tg) expressing different species PRP sequence.
The donor isolate was collected in 1995 in a French commercial sheep flock. inoculation into AHQ/AHQ sheep induced a disease which had all neuro-pathological and biochemical characteristics of atypical scrapie. Transmitted into Transgenic mice expressing either ovine or PrPc, the isolate retained all the described characteristics of atypical scrapie.
Surprisingly the TSE agent characteristics were dramatically different v/hen passaged into Tg bovine mice. The recovered TSE agent had biological and biochemical characteristics similar to those of atypical BSE L in the same mouse model. Moreover, whereas no other TSE agent than BSE were shown to transmit into Tg porcine mice, atypical scrapie was able to develop into this model, albeit with low attack rate on first passage.
Furthermore, after adaptation in the porcine mouse model this prion showed similar biological and biochemical characteristics than BSE adapted to this porcine mouse model. Altogether these data indicate.
(i) the unsuspected potential abilities of atypical scrapie to cross species barriers
(ii) the possible capacity of this agent to acquire new characteristics when crossing species barrier
These findings raise some interrogation on the concept of TSE strain and on the origin of the diversity of the TSE agents and could have consequences on field TSE control measures.
Sunday, March 28, 2010
Nor-98 atypical Scrapie, atypical BSE, spontaneous TSE, trade policy, sound science ?
Monday, December 14, 2009
Similarities between Forms of Sheep Scrapie and Creutzfeldt-Jakob Disease Are Encoded by Distinct Prion Types
(hmmm, this is getting interesting now...TSS) Sporadic CJD type 1 and atypical/ Nor98 scrapie are characterized by fine (reticular) deposits, see also ; All of the Heidenhain variants were of the methionine/ methionine type 1 molecular subtype.
see full text ;
Monday, December 14, 2009
Similarities between Forms of Sheep Scrapie and Creutzfeldt-Jakob Disease Are Encoded by Distinct Prion Types
Monday, February 24, 2014
Sporadic Fatal Insomnia in an Adolescent
To the best of our knowledge, this is the first case of CJD combined with Lewy body disease and argirophilic grain disease. Furthermore, we believe this case is an extremely rare combination of MM2-cortical-type and MM2-thalamic-type sporadic CJD (sCJD), which explains the broad spectrum of MM2-type sCJD findings and symptoms. Moreover, histological features of possible Alzheimer's disease were also reported.
Wednesday, September 03, 2014
Coexistence of mixed phenotype Creutzfeldt-Jakob disease, Lewy body disease and argyrophilic grain disease plus histological features of possible Alzheimer's disease: A multi-protein disorder in an autopsy case
====================================
The familial mutations, Gajdusek proposed, lowered the barrier to such accidental conversion. "Thus," he wrote in 1996, "with these mutations, this ordinarily rare event becomes a ... dominant inherited trait." But Weissmann's qualification still remained to be refuted: the mutations might simply allow easier entry to a lurking virus. ...page 202 Deadly Feast
===================================
something to think about for sure.
but i interpret this as (1st not the gold standard, just my opinion;-), as because of certain gene mutations, one or a family, would be more susceptible to the many different strains of TSE, and the many different proven routes and sources, (which will cause different symptoms, different incubation periods from onset of clinical symptoms to death, different parts of the brain infected, etc.). in other words, it's NOT the gene mutation that CAUSES the disease, but the fact that it makes you more SUSCEPTIBLE, to the TSEs from the surrounding environment, and PLUS accumulation, i think this plays a critical role. maybe there is a one dose scenario, but i think there is more of the 'accumulators' that go clinical, than the 'one dose'. and what is the threshold to sub-clinical to clinical ?
anyway, just pondering out loud here.
also, for anyone interested, there are some studies with links to follow here ;
Friday, January 10, 2014
vpspr, sgss, sffi, TSE, an iatrogenic by-product of gss, ffi, familial type prion disease, what it ???
Monday, August 9, 2010
Variably protease-sensitive prionopathy: A new sporadic disease of the prion protein or just more Prionbaloney ?
Wednesday, March 28, 2012
VARIABLY PROTEASE-SENSITVE PRIONOPATHY IS TRANSMISSIBLE ...price of prion poker goes up again $
OR-10 15:25 - 15:40 VARIABLY PROTEASE-SENSITIVE PRIONOPATHY IS TRANSMISSIBLE IN BANK VOLES Nonno
Tuesday, March 20, 2018
Variably protease-sensitive prionopathy (VPSPr), sporadic creutzfeldt jakob disease sCJD, the same disease, what if?
***2018***
Cervid to human prion transmission
Kong, Qingzhong
Case Western Reserve University, Cleveland, OH, United States
Abstract
Prion disease is transmissible and invariably fatal. Chronic wasting disease (CWD) is the prion disease affecting deer, elk and moose, and it is a widespread and expanding epidemic affecting 22 US States and 2 Canadian provinces so far. CWD poses the most serious zoonotic prion transmission risks in North America because of huge venison consumption (>6 million deer/elk hunted and consumed annually in the USA alone), significant prion infectivity in muscles and other tissues/fluids from CWD-affected cervids, and usually high levels of individual exposure to CWD resulting from consumption of the affected animal among often just family and friends. However, we still do not know whether CWD prions can infect humans in the brain or peripheral tissues or whether clinical/asymptomatic CWD zoonosis has already occurred, and we have no essays to reliably detect CWD infection in humans.
We hypothesize that:
(1) The classic CWD prion strain can infect humans at low levels in the brain and peripheral lymphoid tissues;
(2) The cervid-to-human transmission barrier is dependent on the cervid prion strain and influenced by the host (human) prion protein (PrP) primary sequence;
(3) Reliable essays can be established to detect CWD infection in humans; and
(4) CWD transmission to humans has already occurred. We will test these hypotheses in 4 Aims using transgenic (Tg) mouse models and complementary in vitro approaches.
Aim 1 will prove that the classical CWD strain may infect humans in brain or peripheral lymphoid tissues at low levels by conducting systemic bioassays in a set of humanized Tg mouse lines expressing common human PrP variants using a number of CWD isolates at varying doses and routes. Experimental human CWD samples will also be generated for Aim 3.
Aim 2 will test the hypothesis that the cervid-to-human prion transmission barrier is dependent on prion strain and influenced by the host (human) PrP sequence by examining and comparing the transmission efficiency and phenotypes of several atypical/unusual CWD isolates/strains as well as a few prion strains from other species that have adapted to cervid PrP sequence, utilizing the same panel of humanized Tg mouse lines as in Aim 1.
Aim 3 will establish reliable essays for detection and surveillance of CWD infection in humans by examining in details the clinical, pathological, biochemical and in vitro seeding properties of existing and future experimental human CWD samples generated from Aims 1-2 and compare them with those of common sporadic human Creutzfeldt-Jakob disease (sCJD) prions.
Aim 4 will attempt to detect clinical CWD-affected human cases by examining a significant number of brain samples from prion-affected human subjects in the USA and Canada who have consumed venison from CWD-endemic areas utilizing the criteria and essays established in Aim 3. The findings from this proposal will greatly advance our understandings on the potential and characteristics of cervid prion transmission in humans, establish reliable essays for CWD zoonosis and potentially discover the first case(s) of CWD infection in humans.
Public Health Relevance
There are significant and increasing human exposure to cervid prions because chronic wasting disease (CWD, a widespread and highly infectious prion disease among deer and elk in North America) continues spreading and consumption of venison remains popular, but our understanding on cervid-to-human prion transmission is still very limited, raising public health concerns. This proposal aims to define the zoonotic risks of cervid prions and set up and apply essays to detect CWD zoonosis using mouse models and in vitro methods. The findings will greatly expand our knowledge on the potentials and characteristics of cervid prion transmission in humans, establish reliable essays for such infections and may discover the first case(s) of CWD infection in humans.
Funding Agency
Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS088604-04
Application #
9517118
Study Section
Cellular and Molecular Biology of Neurodegeneration Study Section (CMND)
Program Officer Wong, May
Project Start 2015-09-30 Project End 2019-07-31 Budget Start 2018-08-01 Budget End 2019-07-31 Support Year 4 Fiscal Year 2018 Total Cost Indirect Cost Institution Name Case Western Reserve University Department Pathology Type Schools of Medicine DUNS # 077758407 City Cleveland State OH Country United States Zip Code 44106
Related projects
NIH 2018 R01 NS Cervid to human prion transmission Kong, Qingzhong / Case Western Reserve University
NIH 2017 R01 NS Cervid to human prion transmission Kong, Qingzhong / Case Western Reserve University
NIH 2016 R01 NS Cervid to human prion transmission Kong, Qingzhong / Case Western Reserve University
NIH 2015 R01 NS Cervid to human prion transmission Kong, Qingzhong / Case Western Reserve University $337,507
ZOONOTIC CHRONIC WASTING DISEASE CWD TSE PRION UPDATE
here is the latest;
PRION 2018 CONFERENCE
Oral transmission of CWD into Cynomolgus macaques: signs of atypical disease, prion conversion and infectivity in macaques and bio-assayed transgenic mice
Hermann M. Schatzl, Samia Hannaoui, Yo-Ching Cheng, Sabine Gilch (Calgary Prion Research Unit, University of Calgary, Calgary, Canada) Michael Beekes (RKI Berlin), Walter Schulz-Schaeffer (University of Homburg/Saar, Germany), Christiane Stahl-Hennig (German Primate Center) & Stefanie Czub (CFIA Lethbridge). To date, BSE is the only example of interspecies transmission of an animal prion disease into humans. The potential zoonotic transmission of CWD is an alarming issue and was addressed by many groups using a variety of in vitro and in vivo experimental systems. Evidence from these studies indicated a substantial, if not absolute, species barrier, aligning with the absence of epidemiological evidence suggesting transmission into humans. Studies in non-human primates were not conclusive so far, with oral transmission into new-world monkeys and no transmission into old-world monkeys. Our consortium has challenged 18 Cynomolgus macaques with characterized CWD material, focusing on oral transmission with muscle tissue. Some macaques have orally received a total of 5 kg of muscle material over a period of 2 years.
After 5-7 years of incubation time some animals showed clinical symptoms indicative of prion disease, and prion neuropathology and PrPSc deposition were detected in spinal cord and brain of some euthanized animals. PrPSc in immunoblot was weakly detected in some spinal cord materials and various tissues tested positive in RT-QuIC, including lymph node and spleen homogenates. To prove prion infectivity in the macaque tissues, we have intracerebrally inoculated 2 lines of transgenic mice, expressing either elk or human PrP. At least 3 TgElk mice, receiving tissues from 2 different macaques, showed clinical signs of a progressive prion disease and brains were positive in immunoblot and RT-QuIC. Tissues (brain, spinal cord and spleen) from these and pre-clinical mice are currently tested using various read-outs and by second passage in mice. Transgenic mice expressing human PrP were so far negative for clear clinical prion disease (some mice >300 days p.i.). In parallel, the same macaque materials are inoculated into bank voles.
Taken together, there is strong evidence of transmissibility of CWD orally into macaques and from macaque tissues into transgenic mouse models, although with an incomplete attack rate.
The clinical and pathological presentation in macaques was mostly atypical, with a strong emphasis on spinal cord pathology.
Our ongoing studies will show whether the transmission of CWD into macaques and passage in transgenic mice represents a form of non-adaptive prion amplification, and whether macaque-adapted prions have the potential to infect mice expressing human PrP.
The notion that CWD can be transmitted orally into both new-world and old-world non-human primates asks for a careful reevaluation of the zoonotic risk of CWD..
***> The notion that CWD can be transmitted orally into both new-world and old-world non-human primates asks for a careful reevaluation of the zoonotic risk of CWD. <***
READING OVER THE PRION 2018 ABSTRACT BOOK, LOOKS LIKE THEY FOUND THAT from this study ;
P190 Human prion disease mortality rates by occurrence of chronic wasting disease in freeranging cervids, United States
Abrams JY (1), Maddox RA (1), Schonberger LB (1), Person MK (1), Appleby BS (2), Belay ED (1) (1) Centers for Disease Control and Prevention (CDC), National Center for Emerging and Zoonotic Infectious Diseases, Atlanta, GA, USA (2) Case Western Reserve University, National Prion Disease Pathology Surveillance Center (NPDPSC), Cleveland, OH, USA..
SEEMS THAT THEY FOUND Highly endemic states had a higher rate of prion disease mortality compared to non-CWD states.
AND ANOTHER STUDY;
P172 Peripheral Neuropathy in Patients with Prion Disease
Wang H(1), Cohen M(1), Appleby BS(1,2) (1) University Hospitals Cleveland Medical Center, Cleveland, Ohio (2) National Prion Disease Pathology Surveillance Center, Cleveland, Ohio..
IN THIS STUDY, THERE WERE autopsy-proven prion cases from the National Prion Disease Pathology Surveillance Center that were diagnosed between September 2016 to March 2017,
AND
included 104 patients. SEEMS THEY FOUND THAT The most common sCJD subtype was MV1-2 (30%), followed by MM1-2 (20%),
AND
THAT The Majority of cases were male (60%), AND half of them had exposure to wild game.
snip...see more on Prion 2017 Macaque study from Prion 2017 Conference and other updated science on cwd tse prion zoonosis below...terry
just out CDC...see;
Research Susceptibility of Human Prion Protein to Conversion by Chronic Wasting Disease Prions
Marcelo A. Barria
Adriana Libori, Gordon Mitchell, and Mark W. Head Author affiliations: National CJD Research and Surveillance Unit, University of Edinburgh, Edinburgh, Scotland, UK (M.A. Barria, A. Libori, M.W. Head); National and OIE Reference Laboratory for Scrapie and CWD, Canadian Food Inspection Agency, Ottawa, Ontario, Canada (G. Mitchell) M. A. Barria et al.
ABSTRACT
Chronic wasting disease (CWD) is a contagious and fatal neurodegenerative disease and a serious animal health issue for deer and elk in North America. The identification of the first cases of CWD among free-ranging reindeer and moose in Europe brings back into focus the unresolved issue of whether CWD can be zoonotic like bovine spongiform encephalopathy. We used a cell-free seeded protein misfolding assay to determine whether CWD prions from elk, white-tailed deer, and reindeer in North America can convert the human prion protein to the disease-associated form.
We found that prions can convert, but the efficiency of conversion is affected by polymorphic variation in the cervid and human prion protein genes. In view of the similarity of reindeer, elk, and white-tailed deer in North America to reindeer, red deer, and roe deer, respectively, in Europe, a more comprehensive and thorough assessment of the zoonotic potential of CWD might be warranted.
Molecular Barriers to Zoonotic Transmission of Prions
Marcelo A. Barria, Aru Balachandran, Masanori Morita, Tetsuyuki Kitamoto, Rona Barron, Jean Manson, Richard Knight, James W. Ironside, and Mark W. Headcorresponding author
snip...
The conversion of human PrPC by CWD brain homogenate in PMCA reactions was less efficient when the amino acid at position 129 was valine rather than methionine.
***Furthermore, the form of human PrPres produced in this in vitro assay when seeded with CWD, resembles that found in the most common human prion disease, namely sCJD of the MM1 subtype.
snip...
However, we can say with confidence that under the conditions used here, none of the animal isolates tested were as efficient as C-type BSE in converting human PrPC, which is reassuring.
***Less reassuring is the finding that there is no absolute barrier to the conversion of human PrPC by CWD prions in a protocol using a single round of PMCA and an entirely human substrate prepared from the target organ of prion diseases, the brain.
Prion 2017 Conference Abstracts
CWD 2017 PRION CONFERENCE
First evidence of intracranial and peroral transmission of Chronic Wasting Disease (CWD) into Cynomolgus macaques: a work in progress
Stefanie Czub1, Walter Schulz-Schaeffer2, Christiane Stahl-Hennig3, Michael Beekes4, Hermann Schaetzl5 and Dirk Motzkus6 1 University of Calgary Faculty of Veterinary Medicine/Canadian Food Inspection Agency; 2Universitatsklinikum des Saarlandes und Medizinische Fakultat der Universitat des Saarlandes; 3 Deutsches Primaten Zentrum/Goettingen; 4 Robert-Koch-Institut Berlin; 5 University of Calgary Faculty of Veterinary Medicine; 6 presently: Boehringer Ingelheim Veterinary Research Center; previously: Deutsches Primaten Zentrum/Goettingen
This is a progress report of a project which started in 2009.
21 cynomolgus macaques were challenged with characterized CWD material from white-tailed deer (WTD) or elk by intracerebral (ic), oral, and skin exposure routes.
Additional blood transfusion experiments are supposed to assess the CWD contamination risk of human blood product.
Challenge materials originated from symptomatic cervids for ic, skin scarification and partially per oral routes (WTD brain).
Challenge material for feeding of muscle derived from preclinical WTD and from preclinical macaques for blood transfusion experiments.
We have confirmed that the CWD challenge material contained at least two different CWD agents (brain material) as well as CWD prions in muscle-associated nerves.
Here we present first data on a group of animals either challenged ic with steel wires or per orally and sacrificed with incubation times ranging from 4.5 to 6.9 years at postmortem.
Three animals displayed signs of mild clinical disease, including anxiety, apathy, ataxia and/or tremor. In four animals wasting was observed, two of those had confirmed diabetes.
All animals have variable signs of prion neuropathology in spinal cords and brains and by supersensitive IHC, reaction was detected in spinal cord segments of all animals.
Protein misfolding cyclic amplification (PMCA), real-time quaking-induced conversion (RT-QuiC) and PET-blot assays to further substantiate these findings are on the way, as well as bioassays in bank voles and transgenic mice.
At present, a total of 10 animals are sacrificed and read-outs are ongoing.
Preclinical incubation of the remaining macaques covers a range from 6.4 to 7.10 years.
Based on the species barrier and an incubation time of > 5 years for BSE in macaques and about 10 years for scrapie in macaques, we expected an onset of clinical disease beyond 6 years post inoculation.
PRION 2017
DECIPHERING NEURODEGENERATIVE DISORDERS
Subject: PRION 2017 CONFERENCE
DECIPHERING NEURODEGENERATIVE DISORDERS
VIDEO PRION 2017 CONFERENCE DECIPHERING NEURODEGENERATIVE DISORDERS
*** PRION 2017 CONFERENCE VIDEO
ZOONOTIC, ZOONOSIS, CHRONIC WASTING DISEASE CWD TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY TSE PRION
10. ZOONOTIC, ZOONOSIS, CHRONIC WASTING DISEASE CWD TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY TSE PRION AKA MAD DEER ELK DISEASE IN HUMANS, has it already happened, that should be the question...
''In particular the US data do not clearly exclude the possibility of human (sporadic or familial) TSE development due to consumption of venison. The Working Group thus recognizes a potential risk to consumers if a TSE would be present in European cervids.'' Scientific opinion on chronic wasting disease (II)
EFSA Panel on Biological Hazards (BIOHAZ) Antonia Ricci Ana Allende Declan Bolton Marianne Chemaly Robert Davies Pablo Salvador Fernández Escámez ... See all authors
First published: 17 January 2018 https://doi.org/10.2903/j.efsa.2018.5132 ;
also, see;
8. Even though human TSE‐exposure risk through consumption of game from European cervids can be assumed to be minor, if at all existing, no final conclusion can be drawn due to the overall lack of scientific data. In particular the US data do not clearly exclude the possibility of human (sporadic or familial) TSE development due to consumption of venison. The Working Group thus recognizes a potential risk to consumers if a TSE would be present in European cervids. It might be prudent considering appropriate measures to reduce such a risk, e.g. excluding tissues such as CNS and lymphoid tissues from the human food chain, which would greatly reduce any potential risk for consumers.. However, it is stressed that currently, no data regarding a risk of TSE infections from cervid products are available.
snip...
The tissue distribution of infectivity in CWD‐infected cervids is now known to extend beyond CNS and lymphoid tissues. While the removal of these specific tissues from the food chain would reduce human dietary exposure to infectivity, exclusion from the food chain of the whole carcass of any infected animal would be required to eliminate human dietary exposure.
zoonosis zoonotic cervid tse prion cwd to humans, preparing for the storm
***An alternative to modeling the species barrier is the cell-free conversion assay which points to CWD as the animal prion disease with the greatest zoonotic potential, after (and very much less than) BSE..116***
To date there is no direct evidence that CWD has been or can be transmitted from animals to humans.
However, initial findings from a laboratory research project funded by the Alberta Prion Research Institute (APRI) and Alberta Livestock Meat Agency (ALMA), and led by a Canadian Food Inspection Agency (CFIA) scientist indicate that CWD has been transmitted to cynomolgus macaques (the non-human primate species most closely related to humans that may be used in research), through both the intracranial and oral routes of exposure.
Both infected brain and muscle tissues were found to transmit disease.
Health Canada’s Health Products and Food Branch (HPFB) was asked to consider the impact of these findings on the Branch’s current position on CWD in health products and foods.
Summary and Recommendation:
snip...
Health Portfolio partners were recently made aware of initial findings from a research project led by a CFIA scientist that have demonstrated that cynomolgus macaques can be infected via intracranial exposure and oral gavage with CWD infected muscle.
These findings suggest that CWD, under specific experimental conditions, has the potential to cross the human species barrier, including by enteral feeding of CWD infected muscle.
*** WDA 2016 NEW YORK ***
We found that CWD adapts to a new host more readily than BSE and that human PrP was unexpectedly prone to misfolding by CWD prions.
In addition, we investigated the role of specific regions of the bovine, deer and human PrP protein in resistance to conversion by prions from another species.
***We have concluded that the human protein has a region that confers unusual susceptibility to conversion by CWD prions.
Student Presentations Session 2
The species barriers and public health threat of CWD and BSE prions
Ms. Kristen Davenport1, Dr. Davin Henderson1, Dr. Candace Mathiason1, Dr. Edward Hoover1 1Colorado State University
Chronic wasting disease (CWD) is spreading rapidly through cervid populations in the USA. Bovine spongiform encephalopathy (BSE, mad cow disease) arose in the 1980s because cattle were fed recycled animal protein.
These and other prion diseases are caused by abnormal folding of the normal prion protein (PrP) into a disease causing form (PrPd), which is pathogenic to nervous system cells and can cause subsequent PrP to misfold. CWD spreads among cervids very efficiently, but it has not yet infected humans. On the other hand, BSE was spread only when cattle consumed infected bovine or ovine tissue, but did infect humans and other species.
The objective of this research is to understand the role of PrP structure in cross-species infection by CWD and BSE. To study the propensity of each species’ PrP to be induced to misfold by the presence of PrPd from verious species, we have used an in vitro system that permits detection of PrPd in real-time.
We measured the conversion efficiency of various combinations of PrPd seeds and PrP substrate combinations.
We observed the cross-species behavior of CWD and BSE, in addition to feline-adapted CWD and BSE. We found that CWD adapts to a new host more readily than BSE and that human PrP was unexpectedly prone to misfolding by CWD prions. In addition, we investigated the role of specific regions of the bovine, deer and human PrP protein in resistance to conversion by prions from another species.
***We have concluded that the human protein has a region that confers unusual susceptibility to conversion by CWD prions. CWD is unique among prion diseases in its rapid spread in natural populations. BSE prions are essentially unaltered upon passage to a new species, while CWD adapts to the new species. This adaptation has consequences for surveillance of humans exposed to CWD. Wildlife Disease Risk Communication Research Contributes to Wildlife Trust Administration Exploring perceptions about chronic wasting disease risks among wildlife and agriculture professionals and stakeholders
TUESDAY, SEPTEMBER 12, 2017
CDC Now Recommends Strongly consider having the deer or elk tested for CWD before you eat the meat
SATURDAY, JANUARY 27, 2018
CDC CHRONIC WASTING DISEASE CWD TSE PRION UPDATE REPORT USA JANUARY 2018
Subject: CDC CHRONIC WASTING DISEASE CWD TSE PRION UPDATE REPORT USA JANUARY 2018
CHRONIC WASTING DISEASE CWD TSE PRION IS THE USA AND NORTH AMERICA'S MAD COW DISEASE.
THE USDA INC ET AL WORKED VERY HARD CONCEALING BSE TSE PRION IN CATTLE. they almost succeeded $$$
BUT CWD TSE PRION IN CERVIDS IS A DIFFERENT BEAST, THE COVER UP THERE, USDA INC COULD NOT CONTAIN.
SPORADIC CJD IS 85%+ OF ALL HUMAN TSE PRION DISEASE.
SPORADIC CJD HAS NOW BEEN LINKED TO TYPICAL AND ATYPICAL BSE, SCRAPIE, AND CWD.
SPORADIC/SPONTANEOUS TSE HAS NEVER BEEN PROVEN.
***Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility.***
CDC CWD TSE PRION UPDATE USA JANUARY 2018
As of January 2018, CWD in free-ranging deer, elk and/or moose has been reported in at least 22 states in the continental United States, as well as two provinces in Canada. In addition, CWD has been reported in reindeer and moose in Norway, and a small number of imported cases have been reported in South Korea. The disease has also been found in farmed deer and elk. CWD was first identified in captive deer in the late 1960s in Colorado and in wild deer in 1981. By the 1990s, it had been reported in surrounding areas in northern Colorado and southern Wyoming. Since 2000, the area known to be affected by CWD in free-ranging animals has increased to at least 22 states, including states in the Midwest, Southwest, and limited areas on the East Coast.. It is possible that CWD may also occur in other states without strong animal surveillance systems, but that cases haven’t been detected yet. Once CWD is established in an area, the risk can remain for a long time in the environment. The affected areas are likely to continue to expand. Nationwide, the overall occurrence of CWD in free-ranging deer and elk is relatively low. However, in several locations where the disease is established, infection rates may exceed 10 percent (1 in 10), and localized infection rates of more than 25 percent (1 in 4) have been reported. The infection rates among some captive deer can be much higher, with a rate of 79% (nearly 4 in 5) reported from at least one captive herd. As of January 2018, there were 186 counties in 22 states with reported CWD in free-ranging cervids...
Chronic Wasting Disease Among Free-Ranging Cervids by County, United States, January 2018
snip....
*** 2017-2018 CWD TSE Prion UPDATE
*** The potential impact of prion diseases on human health was greatly magnified by the recognition that interspecies transfer of BSE to humans by beef ingestion resulted in vCJD. While changes in animal feed constituents and slaughter practices appear to have curtailed vCJD, there is concern that CWD of free-ranging deer and elk in the U.S. might also cross the species barrier. Thus, consuming venison could be a source of human prion disease. Whether BSE and CWD represent interspecies scrapie transfer or are newly arisen prion diseases is unknown. Therefore, the possibility of transmission of prion disease through other food animals cannot be ruled out. There is evidence that vCJD can be transmitted through blood transfusion. There is likely a pool of unknown size of asymptomatic individuals infected with vCJD, and there may be asymptomatic individuals infected with the CWD equivalent. These circumstances represent a potential threat to blood, blood products, and plasma supplies.
Transmission Studies
Mule deer transmissions of CWD were by intracerebral inoculation and compared with natural cases {the following was written but with a single line marked through it ''first passage (by this route)}....TSS
resulted in a more rapidly progressive clinical disease with repeated episodes of synocopy ending in coma. One control animal became affected, it is believed through contamination of inoculum (?saline). Further CWD transmissions were carried out by Dick Marsh into ferret, mink and squirrel monkey. Transmission occurred in ALL of these species with the shortest incubation period in the ferret.
snip....
Prion Infectivity in Fat of Deer with Chronic Wasting Disease▿
Brent Race#, Kimberly Meade-White#, Richard Race and Bruce Chesebro* + Author Affiliations
In mice, prion infectivity was recently detected in fat. Since ruminant fat is consumed by humans and fed to animals, we determined infectivity titers in fat from two CWD-infected deer. Deer fat devoid of muscle contained low levels of CWD infectivity and might be a risk factor for prion infection of other species.
Prions in Skeletal Muscles of Deer with Chronic Wasting Disease
Here bioassays in transgenic mice expressing cervid prion protein revealed the presence of infectious prions in skeletal muscles of CWD-infected deer, demonstrating that humans consuming or handling meat from CWD-infected deer are at risk to prion exposure.
*** now, let’s see what the authors said about this casual link, personal communications years ago, and then the latest on the zoonotic potential from CWD to humans from the TOKYO PRION 2016 CONFERENCE.
see where it is stated NO STRONG evidence. so, does this mean there IS casual evidence ???? “Our conclusion stating that we found no strong evidence of CWD transmission to humans”
From: TSS (216-119-163-189.ipset45.wt.net)
Subject: CWD aka MAD DEER/ELK TO HUMANS ???
Date: September 30, 2002 at 7:06 am PST
From: "Belay, Ermias"
To: Cc: "Race, Richard (NIH)" ; ; "Belay, Ermias"
Sent: Monday, September 30, 2002 9:22 AM
Subject: RE: TO CDC AND NIH - PUB MED- 3 MORE DEATHS - CWD - YOUNG HUNTERS
Dear Sir/Madam,
In the Archives of Neurology you quoted (the abstract of which was attached to your email), we did not say CWD in humans will present like variant CJD.. That assumption would be wrong. I encourage you to read the whole article and call me if you have questions or need more clarification (phone: 404-639-3091). Also, we do not claim that "no-one has ever been infected with prion disease from eating venison." Our conclusion stating that we found no strong evidence of CWD transmission to humans in the article you quoted or in any other forum is limited to the patients we investigated.
Ermias Belay, M.D. Centers for Disease Control and Prevention
-----Original Message-----
From: Sent: Sunday, September 29, 2002 10:15 AM
Subject: TO CDC AND NIH - PUB MED- 3 MORE DEATHS - CWD - YOUNG HUNTERS
Sunday, November 10, 2002 6:26 PM .......snip........end..............TSS
Thursday, April 03, 2008
A prion disease of cervids: Chronic wasting disease 2008 1: Vet Res. 2008 Apr 3;39(4):41 A prion disease of cervids: Chronic wasting disease Sigurdson CJ.
snip...
*** twenty-seven CJD patients who regularly consumed venison were reported to the Surveillance Center***,
snip... full text ;
> However, to date, no CWD infections have been reported in people.
key word here is 'reported'. science has shown that CWD in humans will look like sporadic CJD. SO, how can one assume that CWD has not already transmitted to humans? they can't, and it's as simple as that. from all recorded science to date, CWD has already transmitted to humans, and it's being misdiagnosed as sporadic CJD. ...terry
*** LOOKING FOR CWD IN HUMANS AS nvCJD or as an ATYPICAL CJD, LOOKING IN ALL THE WRONG PLACES $$$ ***
*** These results would seem to suggest that CWD does indeed have zoonotic potential, at least as judged by the compatibility of CWD prions and their human PrPC target. Furthermore, extrapolation from this simple in vitro assay suggests that if zoonotic CWD occurred, it would most likely effect those of the PRNP codon 129-MM genotype and that the PrPres type would be similar to that found in the most common subtype of sCJD (MM1).***
SEE; Travel History, Hunting, and Venison Consumption Related to Prion Disease Exposure, 2006-2007 FoodNet Population Survey
Monday, May 23, 2011
CDC Assesses Potential Human Exposure to Prion Diseases Travel Warning
Public release date: 23-May-2011
Contact: Francesca Costanzo adajmedia@elsevier.com 215-239-3249 Elsevier Health Sciences
CDC assesses potential human exposure to prion diseases Study results reported in the Journal of the American Dietetic Association
Philadelphia, PA, May 23, 2011 – Researchers from the Centers for Disease Control and Prevention (CDC) have examined the potential for human exposure to prion diseases, looking at hunting, venison consumption, and travel to areas in which prion diseases have been reported in animals. Three prion diseases in particular – bovine spongiform encephalopathy (BSE or “Mad Cow Disease”), variant Creutzfeldt-Jakob disease (vCJD), and chronic wasting disease (CWD) – were specified in the investigation. The results of this investigation are published in the June issue of the Journal of the American Dietetic Association.
“While prion diseases are rare, they are generally fatal for anyone who becomes infected. More than anything else, the results of this study support the need for continued surveillance of prion diseases,” commented lead investigator Joseph Y. Abrams, MPH, National Center for Emerging and Zoonotic Infectious Diseases, CDC, Atlanta.”But it’s also important that people know the facts about these diseases, especially since this study shows that a good number of people have participated in activities that may expose them to infection-causing agents.”
Although rare, human prion diseases such as CJD may be related to BSE. Prion (proteinaceous infectious particles) diseases are a group of rare brain diseases that affect humans and animals. When a person gets a prion disease, brain function is impaired. This causes memory and personality changes, dementia, and problems with movement. All of these worsen over time. These diseases are invariably fatal. Since these diseases may take years to manifest, knowing the extent of human exposure to possible prion diseases could become important in the event of an outbreak.
CDC investigators evaluated the results of the 2006-2007 population survey conducted by the Foodborne Diseases Active Surveillance Network (FoodNet). This survey collects information on food consumption practices, health outcomes, and demographic characteristics of residents of the participating Emerging Infections Program sites. The survey was conducted in Connecticut, Georgia, Maryland, Minnesota, New Mexico, Oregon, and Tennessee, as well as five counties in the San Francisco Bay area, seven counties in the Greater Denver area, and 34 counties in western and northeastern New York.
Survey participants were asked about behaviors that could be associated with exposure to the agents causing BSE and CWD, including travel to the nine countries considered to be BSE-endemic (United Kingdom, Republic of Ireland, France, Portugal, Switzerland, Italy, the Netherlands, Germany, Spain) and the cumulative length of stay in each of those countries. Respondents were asked if they ever had hunted for deer or elk, and if that hunting had taken place in areas considered to be CWD-endemic (northeastern Colorado, southeastern Wyoming or southwestern Nebraska). They were also asked if they had ever consumed venison, the frequency of consumption, and whether the meat came from the wild.
The proportion of survey respondents who reported travel to at least one of the nine BSE endemic countries since 1980 was 29.5%. Travel to the United Kingdom was reported by 19.4% of respondents, higher than to any other BSE-endemic country. Among those who traveled, the median duration of travel to the United Kingdom (14 days) was longer than that of any other BSE-endemic country.. Travelers to the UK were more likely to have spent at least 30 days in the country (24.9%) compared to travelers to any other BSE endemic country. The prevalence and extent of travel to the UK indicate that health concerns in the UK may also become issues for US residents.
The proportion of survey respondents reporting having hunted for deer or elk was 18.5% and 1.2% reported having hunted for deer or elk in CWD-endemic areas. Venison consumption was reported by 67.4% of FoodNet respondents, and 88.6% of those reporting venison consumption had obtained all of their meat from the wild. These findings reinforce the importance of CWD surveillance and control programs for wild deer and elk to reduce human exposure to the CWD agent. Hunters in CWD-endemic areas are advised to take simple precautions such as: avoiding consuming meat from sickly deer or elk, avoiding consuming brain or spinal cord tissues, minimizing the handling of brain and spinal cord tissues, and wearing gloves when field-dressing carcasses.
According to Abrams, “The 2006-2007 FoodNet population survey provides useful information should foodborne prion infection become an increasing public health concern in the future. The data presented describe the prevalence of important behaviors and their associations with demographic characteristics. Surveillance of BSE, CWD, and human prion diseases are critical aspects of addressing the burden of these diseases in animal populations and how that may relate to human health.”
###
The article is “Travel history, hunting, and venison consumption related to prion disease exposure, 2006-2007 FoodNet population survey” by Joseph Y. Abrams, MPH; Ryan A. Maddox, MPH; Alexis R Harvey, MPH; Lawrence B. Schonberger, MD; and Ermias D. Belay, MD. It appears in the Journal of the American Dietetic Association, Volume 111, Issue 6 (June 2011) published by Elsevier.
In an accompanying podcast CDC’s Joseph Y. Abrams discusses travel, hunting, and eating venison in relation to prion diseases. It is available at http://adajournal.org/content/podcast. ;
Thursday, May 26, 2011
Travel History, Hunting, and Venison Consumption Related to Prion Disease Exposure, 2006-2007 FoodNet Population Survey
Journal of the American Dietetic Association Volume 111, Issue 6 , Pages 858-863, June 2011.
Travel History, Hunting, and Venison Consumption Related to Prion Disease Exposure, 2006-2007 FoodNet Population Survey
Joseph Y. Abrams, MPH, Ryan A. Maddox, MPH , Alexis R. Harvey, MPH , Lawrence B. Schonberger, MD , Ermias D. Belay, MD
Accepted 15 November 2010. Abstract Full Text PDF References .
Abstract
The transmission of bovine spongiform encephalopathy (BSE) to human beings and the spread of chronic wasting disease (CWD) among cervids have prompted concerns about zoonotic transmission of prion diseases. Travel to the United Kingdom and other European countries, hunting for deer or elk, and venison consumption could result in the exposure of US residents to the agents that cause BSE and CWD. The Foodborne Diseases Active Surveillance Network 2006-2007 population survey was used to assess the prevalence of these behaviors among residents of 10 catchment areas across the United States. Of 17,372 survey respondents, 19.4% reported travel to the United Kingdom since 1980, and 29.5% reported travel to any of the nine European countries considered to be BSE-endemic since 1980. The proportion of respondents who had ever hunted deer or elk was 18.5%, and 1.2% had hunted deer or elk in a CWD–endemic area. More than two thirds (67.4%) reported having ever eaten deer or elk meat. Respondents who traveled spent more time in the United Kingdom (median 14 days) than in any other BSE-endemic country. Of the 11,635 respondents who had consumed venison, 59.8% ate venison at most one to two times during their year of highest consumption, and 88.6% had obtained all of their meat from the wild. The survey results were useful in determining the prevalence and frequency of behaviors that could be important factors for foodborne prion transmission.
PLUS, THE CDC DID NOT PUT THIS WARNING OUT FOR THE WELL BEING OF THE DEER AND ELK ;
Thursday, May 26, 2011
Travel History, Hunting, and Venison Consumption Related to Prion Disease Exposure, 2006-2007 FoodNet Population Survey
Journal of the American Dietetic Association Volume 111, Issue 6 , Pages 858-863, June 2011.
NOR IS THE FDA recalling this CWD positive elk meat for the well being of the dead elk ;
Wednesday, March 18, 2009
Noah's Ark Holding, LLC, Dawson, MN RECALL Elk products contain meat derived from an elk confirmed to have CWD NV, CA, TX, CO, NY, UT, FL, OK RECALLS AND FIELD CORRECTIONS: FOODS CLASS II
Transmissible Spongiform Encephalopathies
Spongiform Encephalopathy in Captive Wild ZOO BSE INQUIRY
BSE INQUIRY
CJD9/10022
October 1994
Mr R.N. Elmhirst Chairman British Deer Farmers Association Holly Lodge Spencers Lane
BerksWell Coventry CV7 7BZ
Dear Mr Elmhirst,
CREUTZFELDT-JAKOB DISEASE (CJD) SURVEILLANCE UNIT REPORT
Thank you for your recent letter concerning the publication of the third annual report from the CJD Surveillance Unit. I am sorry that you are dissatisfied with the way in which this report was published.
The Surveillance Unit is a completely independant outside body and the Department of Health is committed to publishing their reports as soon as they become available. In the circumstances it is not the practice to circulate the report for comment since the findings of the report would not be amended.. In future we can ensure that the British Deer Farmers Association receives a copy of the report in advance of publication.
The Chief Medical Officer has undertaken to keep the public fully informed of the results of any research in respect of CJD. This report was entirely the work of the unit and was produced completely independantly of the the Department.
The statistical results reqarding the consumption of venison was put into perspective in the body of the report and was not mentioned at all in the press release. Media attention regarding this report was low key but gave a realistic presentation of the statistical findings of the Unit. This approach to publication was successful in that consumption of venison was highlighted only once by the media ie. in the News at one television proqramme.
I believe that a further statement about the report, or indeed statistical links between CJD and consumption of venison, would increase, and quite possibly give damaging credence, to the whole issue. From the low key media reports of which I am aware it seems unlikely that venison consumption will suffer adversely, if at all.
http://web.archive.org/web/20030511010117/http://www.bseinquiry.gov.uk/files/yb/1994/10/00003001.pdf
*** The association between venison eating and risk of CJD shows similar pattern, with regular venison eating associated with a 9 FOLD INCREASE IN RISK OF CJD (p = 0.04). ***
*** The association between venison eating and risk of CJD shows similar pattern, with regular venison eating associated with a 9 FOLD INCREASE IN RISK OF CJD (p = 0.04). ***
*** The association between venison eating and risk of CJD shows similar pattern, with regular venison eating associated with a 9 FOLD INCREASE IN RISK OF CJD (p = 0.04). ***
There is some evidence that risk of CJD INCREASES WITH INCREASING FREQUENCY OF LAMB EATING (p = 0.02)..
The evidence for such an association between beef eating and CJD is weaker (p = 0.14). When only controls for whom a relative was interviewed are included, this evidence becomes a little STRONGER (p = 0.08).
snip...
It was found that when veal was included in the model with another exposure, the association between veal and CJD remained statistically significant (p = < 0.05 for all exposures), while the other exposures ceased to be statistically significant (p = > 0.05).
snip...
In conclusion, an analysis of dietary histories revealed statistical associations between various meats/animal products and INCREASED RISK OF CJD. When some account was taken of possible confounding, the association between VEAL EATING AND RISK OF CJD EMERGED AS THE STRONGEST OF THESE ASSOCIATIONS STATISTICALLY. ...
snip...
In the study in the USA, a range of foodstuffs were associated with an increased risk of CJD, including liver consumption which was associated with an apparent SIX-FOLD INCREASE IN THE RISK OF CJD. By comparing the data from 3 studies in relation to this particular dietary factor, the risk of liver consumption became non-significant with an odds ratio of 1.2 (PERSONAL COMMUNICATION, PROFESSOR A. HOFMAN. ERASMUS UNIVERSITY, ROTTERDAM). (???...TSS)
snip...see full report ;
https://web.archive.org/web/20170126073306/http://collections..europarchive..org/tna/20090505194948/http://bseinquiry.gov.uk/files/yb/1994/08/00004001.pdf
MONDAY, NOVEMBER 26, 2018
***> Wisconsin CWD spreads on deer and elk farms as control efforts stumble
WITH the recent findings that Scrapie will transmit to Macaque by oral route, that Scrapie and CWD TSE Prion will transit to pigs orally, recent outbreak documented of TSE Prion Disease in Dromedary Camels, Algeria, atypical TSE Prion still being documented, and again just recently in the USA, of another atypical BSE case, and this discovery was only documented by testing 20k head of cattle from some 100M head of cattle in any given year in the USA, the continued denial that atypical BSE and atypical Scrapie are a transmissible disease (science has shown otherwise) this is concerning to me. Science and scientific policy makers have forgotten what Gibbs, Gajdusek, Hadlow, Alper, Zigas, even Gordon with the infamous Scrapie vaccine blunder, a discovery of valuable importance, and so many others i am failing to remember now, what some found long ago, like Dr. Gibbs, he tried to warn us about scrapie zoonosis potential, yet that went ignored for decades and decades. we/scientist/officials/the world, knows the USA FDA PART 589 TSE PRION FEED ban has failed terribly, the BSE testing has failed terribly, and the surveillance there from has failed, SRM removal breaches, all proven by the OIG or the GAO, and others. But yet, we find ourselves now debating the issue of these same risk factors for scrapie, the same risk factors that we all knew were there, with science staring us in the face, we still deny scientific facts all in the name of corporate interest. let's not continue to make these same mistakes. human and animal life is at stake here. we must remove corporate/government/lobbyist interest from the scientific policy making and regulations there from for the TSE Prion, all of them. ...Terry S. Singeltary SR.
O3 Experimental studies on prion transmission barrier and TSE pathogenesis in large animals
Rosa Bolea(1), Acín C(1)Marín B(1), Hedman C(1), Raksa H(1), Barrio T(1), Otero A(1), LópezPérez O(1), Monleón E(1),Martín-Burriel(1), Monzón M(1), Garza MC(1), Filali H(1),Pitarch JL(1), Garcés M(1), Betancor M(1), GuijarroIM(1), GarcíaM(1), Moreno B(1),Vargas A(1), Vidal E(2), Pumarola M(2), Castilla J(3), Andréoletti O(4), Espinosa JC(5), Torres JM(5), Badiola JJ(1).
1Centro de Investigación en Encefalopatías y Enfermedades Transmisibles Emergentes, VeterinaryFaculty, Universidad de Zaragoza; Zaragoza,Spain.2 RTA, Centre de Recerca en Sanitat Animal (CReSA, IRTA-UAB) 3 4 INRA, ÉcoleVétérinaire, Toulouse, France.5CIC bioGUNE, Prion researchlab, Derio, Spain CISA- INIA, Valdeolmos, Madrid 28130, Spain.
Experimental transmission of Transmissible Spongiform Encephalopathies (TSE) has been understood and related with several factors that could modify the natural development of these diseases. In fact, the behaviour of the natural disease does not match exactly in each animal, being modified by parameters such as the age at infection, the genotype, the breed or the causative strain. Moreover, different TSE strains can target different animal species or tissues, what complicate the prediction of its transmissibility when is tested in a different species of the origin source. The aim of the experimental studies in large animals is to homogenize all those factors, trying to minimize as much as possible variations between individuals. These effects can be flattened by experimental transmission in mice, in which a specific strain can be selected after several passages. With this objective, several experimental studies in large animals have been developed by the presenter research team.
Classical scrapie agent has been inoculated in cow, with the aim of demonstrate the resistance or susceptibility of this species to the first well known TSE; Atypical scrapie has been inoculated in sheep (using several routes of infection), cow and pig, with the objective of evaluating the potential pathogenicity of this strain; Classical Bovine Spongiform Encephalopathy (BSE) has been inoculated in goats aiming to demonstrate if the genetic background of this species could protect against this strain; goat BSE and sheep BSE have been inoculated in goats and pigs respectively to evaluate the effect of species barrier; and finally atypical BSE has been inoculated in cattle to assess the transmissibility properties of this newly introduced strain.
Once the experiments have been carried out on large animal species, a collection of samples from animals studied were inoculated in different types of tg mice overexpressing PrPcin order to study the infectivity of the tissues, and also were studied using PMCA.
In summary, the parameters that have been controlled are the species, the strain, the route of inoculation, the time at infection, the genotype, the age, and the environmental conditions.
To date,
***> eleven of the atypical scrapie intracerebrally inoculated sheep have succumbed to atypical scrapie disease;
***> six pigs to sheep BSE;
***> one cow to classical scrapie;
***> nine goats to goat BSE and
***> five goats to classical BSE.
***> PrPSC has been demonstrated in all cases by immunohistochemistry and western blot.
=====> PRION CONFERENCE 2018
why do we not want to do TSE transmission studies on chimpanzees $
5. A positive result from a chimpanzee challenged severly would likely create alarm in some circles even if the result could not be interpreted for man.
***> I have a view that all these agents could be transmitted provided a large enough dose by appropriate routes was given and the animals kept long enough.
***> Until the mechanisms of the species barrier are more clearly understood it might be best to retain that hypothesis.
snip...
ZOONOSIS OF SCRAPIE TSE PRION
O.05: Transmission of prions to primates after extended silent incubation periods: Implications for BSE and scrapie risk assessment in human populations
Emmanuel Comoy, Jacqueline Mikol, Valerie Durand, Sophie Luccantoni, Evelyne Correia, Nathalie Lescoutra, Capucine Dehen, and Jean-Philippe Deslys Atomic Energy Commission; Fontenay-aux-Roses, France
Prion diseases (PD) are the unique neurodegenerative proteinopathies reputed to be transmissible under field conditions since decades. The transmission of Bovine Spongiform Encephalopathy (BSE) to humans evidenced that an animal PD might be zoonotic under appropriate conditions. Contrarily, in the absence of obvious (epidemiological or experimental) elements supporting a transmission or genetic predispositions, PD, like the other proteinopathies, are reputed to occur spontaneously (atpical animal prion strains, sporadic CJD summing 80% of human prion cases).
Non-human primate models provided the first evidences supporting the transmissibiity of human prion strains and the zoonotic potential of BSE. Among them, cynomolgus macaques brought major information for BSE risk assessment for human health (Chen, 2014), according to their phylogenetic proximity to humans and extended lifetime. We used this model to assess the zoonotic potential of other animal PD from bovine, ovine and cervid origins even after very long silent incubation periods.
*** We recently observed the direct transmission of a natural classical scrapie isolate to macaque after a 10-year silent incubation period,
***with features similar to some reported for human cases of sporadic CJD, albeit requiring fourfold long incubation than BSE. Scrapie, as recently evoked in humanized mice (Cassard, 2014),
***is the third potentially zoonotic PD (with BSE and L-type BSE),
***thus questioning the origin of human sporadic cases.
We will present an updated panorama of our different transmission studies and discuss the implications of such extended incubation periods on risk assessment of animal PD for human health.
===============
***thus questioning the origin of human sporadic cases***
===============
***our findings suggest that possible transmission risk of H-type BSE to sheep and human. Bioassay will be required to determine whether the PMCA products are infectious to these animals.
==============
***Transmission data also revealed that several scrapie prions propagate in HuPrP-Tg mice with efficiency comparable to that of cattle BSE. While the efficiency of transmission at primary passage was low, subsequent passages resulted in a highly virulent prion disease in both Met129 and Val129 mice.
***Transmission of the different scrapie isolates in these mice leads to the emergence of prion strain phenotypes that showed similar characteristics to those displayed by MM1 or VV2 sCJD prion.
***These results demonstrate that scrapie prions have a zoonotic potential and raise new questions about the possible link between animal and human prions.
PRION 2016 TOKYO
Saturday, April 23, 2016
SCRAPIE WS-01: Prion diseases in animals and zoonotic potential 2016
Prion. 10:S15-S21. 2016 ISSN: 1933-6896 printl 1933-690X online
Taylor & Francis
Prion 2016 Animal Prion Disease Workshop Abstracts
WS-01: Prion diseases in animals and zoonotic potential
Juan Maria Torres a, Olivier Andreoletti b, J uan-Carlos Espinosa a. Vincent Beringue c. Patricia Aguilar a,
Natalia Fernandez-Borges a. and Alba Marin-Moreno a
"Centro de Investigacion en Sanidad Animal ( CISA-INIA ). Valdeolmos, Madrid. Spain; b UMR INRA -ENVT 1225 Interactions Holes Agents Pathogenes. ENVT. Toulouse. France: "UR892. Virologie lmmunologie MolécuIaires, Jouy-en-Josas. France
Dietary exposure to bovine spongiform encephalopathy (BSE) contaminated bovine tissues is considered as the origin of variant Creutzfeldt Jakob (vCJD) disease in human. To date, BSE agent is the only recognized zoonotic prion... Despite the variety of Transmissible Spongiform Encephalopathy (TSE) agents that have been circulating for centuries in farmed ruminants there is no apparent epidemiological link between exposure to ruminant products and the occurrence of other form of TSE in human like sporadic Creutzfeldt Jakob Disease (sCJD). However, the zoonotic potential of the diversity of circulating TSE agents has never been systematically assessed. The major issue in experimental assessment of TSEs zoonotic potential lies in the modeling of the ‘species barrier‘, the biological phenomenon that limits TSE agents’ propagation from a species to another. In the last decade, mice genetically engineered to express normal forms of the human prion protein has proved essential in studying human prions pathogenesis and modeling the capacity of TSEs to cross the human species barrier.
To assess the zoonotic potential of prions circulating in farmed ruminants, we study their transmission ability in transgenic mice expressing human PrPC (HuPrP-Tg). Two lines of mice expressing different forms of the human PrPC (129Met or 129Val) are used to determine the role of the Met129Val dimorphism in susceptibility/resistance to the different agents.
These transmission experiments confirm the ability of BSE prions to propagate in 129M- HuPrP-Tg mice and demonstrate that Met129 homozygotes may be susceptible to BSE in sheep or goat to a greater degree than the BSE agent in cattle and that these agents can convey molecular properties and neuropathological indistinguishable from vCJD. However homozygous 129V mice are resistant to all tested BSE derived prions independently of the originating species suggesting a higher transmission barrier for 129V-PrP variant.
Transmission data also revealed that several scrapie prions propagate in HuPrP-Tg mice with efficiency comparable to that of cattle BSE. While the efficiency of transmission at primary passage was low, subsequent passages resulted in a highly virulent prion disease in both Met129 and Val129 mice.
Transmission of the different scrapie isolates in these mice leads to the emergence of prion strain phenotypes that showed similar characteristics to those displayed by MM1 or VV2 sCJD prion.
These results demonstrate that scrapie prions have a zoonotic potential and raise new questions about the possible link between animal and human prions.
why do we not want to do TSE transmission studies on chimpanzees $
5. A positive result from a chimpanzee challenged severly would likely create alarm in some circles even if the result could not be interpreted for man.
***> I have a view that all these agents could be transmitted provided a large enough dose by appropriate routes was given and the animals kept long enough.
***> Until the mechanisms of the species barrier are more clearly understood it might be best to retain that hypothesis.
snip...
R. BRADLEY
Title: Transmission of scrapie prions to primate after an extended silent incubation period)
*** In complement to the recent demonstration that humanized mice are susceptible to scrapie, we report here the first observation of direct transmission of a natural classical scrapie isolate to a macaque after a 10-year incubation period. Neuropathologic examination revealed all of the features of a prion disease: spongiform change, neuronal loss, and accumulation of PrPres throughout the CNS.
*** This observation strengthens the questioning of the harmlessness of scrapie to humans, at a time when protective measures for human and animal health are being dismantled and reduced as c-BSE is considered controlled and being eradicated.
*** Our results underscore the importance of precautionary and protective measures and the necessity for long-term experimental transmission studies to assess the zoonotic potential of other animal prion strains.
***> Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility. <***
Transmission of scrapie prions to primate after an extended silent incubation period
Emmanuel E. Comoy, Jacqueline Mikol, Sophie Luccantoni-Freire, Evelyne Correia, Nathalie Lescoutra-Etchegaray, Valérie Durand, Capucine Dehen, Olivier Andreoletti, Cristina Casalone, Juergen A. Richt, Justin J. Greenlee, Thierry Baron, Sylvie L. Benestad, Paul Brown & Jean-Philippe Deslys Scientific Reports volume 5, Article number: 11573 (2015) | Download Citation
Abstract
Classical bovine spongiform encephalopathy (c-BSE) is the only animal prion disease reputed to be zoonotic, causing variant Creutzfeldt-Jakob disease (vCJD) in humans and having guided protective measures for animal and human health against animal prion diseases. Recently, partial transmissions to humanized mice showed that the zoonotic potential of scrapie might be similar to c-BSE. We here report the direct transmission of a natural classical scrapie isolate to cynomolgus macaque, a highly relevant model for human prion diseases, after a 10-year silent incubation period, with features similar to those reported for human cases of sporadic CJD. Scrapie is thus actually transmissible to primates with incubation periods compatible with their life expectancy, although fourfold longer than BSE. Long-term experimental transmission studies are necessary to better assess the zoonotic potential of other prion diseases with high prevalence, notably Chronic Wasting Disease of deer and elk and atypical/Nor98 scrapie.
SNIP...
Discussion We describe the transmission of spongiform encephalopathy in a non-human primate inoculated 10 years earlier with a strain of sheep c-scrapie. Because of this extended incubation period in a facility in which other prion diseases are under study, we are obliged to consider two alternative possibilities that might explain its occurrence. We first considered the possibility of a sporadic origin (like CJD in humans). Such an event is extremely improbable because the inoculated animal was 14 years old when the clinical signs appeared, i.e. about 40% through the expected natural lifetime of this species, compared to a peak age incidence of 60–65 years in human sporadic CJD, or about 80% through their expected lifetimes. Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility.
The second possibility is a laboratory cross-contamination. Three facts make this possibility equally unlikely. First, handling of specimens in our laboratory is performed with fastidious attention to the avoidance of any such cross-contamination. Second, no laboratory cross-contamination has ever been documented in other primate laboratories, including the NIH, even between infected and uninfected animals housed in the same or adjacent cages with daily intimate contact (P. Brown, personal communication). Third, the cerebral lesion profile is different from all the other prion diseases we have studied in this model19, with a correlation between cerebellar lesions (massive spongiform change of Purkinje cells, intense PrPres staining and reactive gliosis26) and ataxia. The iron deposits present in the globus pallidus are a non specific finding that have been reported previously in neurodegenerative diseases and aging27. Conversely, the thalamic lesion was reminiscent of a metabolic disease due to thiamine deficiency28 but blood thiamine levels were within normal limits (data not shown). The preferential distribution of spongiform change in cortex associated with a limited distribution in the brainstem is reminiscent of the lesion profile in MM2c and VV1 sCJD patients29, but interspecies comparison of lesion profiles should be interpreted with caution. It is of note that the same classical scrapie isolate induced TSE in C57Bl/6 mice with similar incubation periods and lesional profiles as a sample derived from a MM1 sCJD patient30.
We are therefore confident that the illness in this cynomolgus macaque represents a true transmission of a sheep c-scrapie isolate directly to an old-world monkey, which taxonomically resides in the primate subdivision (parvorder of catarrhini) that includes humans. With an homology of its PrP protein with humans of 96.4%31, cynomolgus macaque constitutes a highly relevant model for assessing zoonotic risk of prion diseases. Since our initial aim was to show the absence of transmission of scrapie to macaques in the worst-case scenario, we obtained materials from a flock of naturally-infected sheep, affecting animals with different genotypes32. This c-scrapie isolate exhibited complete transmission in ARQ/ARQ sheep (332 ± 56 days) and Tg338 transgenic mice expressing ovine VRQ/VRQ prion protein (220 ± 5 days) (O. Andreoletti, personal communication). From the standpoint of zoonotic risk, it is important to note that sheep with c-scrapie (including the isolate used in our study) have demonstrable infectivity throughout their lymphoreticular system early in the incubation period of the disease (3 months-old for all the lymphoid organs, and as early as 2 months-old in gut-associated lymph nodes)33. In addition, scrapie infectivity has been identified in blood34, milk35 and skeletal muscle36 from asymptomatic but scrapie infected small ruminants which implies a potential dietary exposure for consumers.
Two earlier studies have reported the occurrence of clinical TSE in cynomolgus macaques after exposures to scrapie isolates. In the first study, the “Compton” scrapie isolate (derived from an English sheep) and serially propagated for 9 passages in goats did not transmit TSE in cynomolgus macaque, rhesus macaque or chimpanzee within 7 years following intracerebral challenge1; conversely, after 8 supplementary passages in conventional mice, this “Compton” isolate induced TSE in a cynomolgus macaque 5 years after intracerebral challenge, but rhesus macaques and chimpanzee remained asymptomatic 8.5 years post-exposure8. However, multiple successive passages that are classically used to select laboratory-adapted prion strains can significantly modify the initial properties of a scrapie isolate, thus questioning the relevance of zoonotic potential for the initial sheep-derived isolate. The same isolate had also induced disease into squirrel monkeys (new-world monkey)9. A second historical observation reported that a cynomolgus macaque developed TSE 6 years post-inoculation with brain homogenate from a scrapie-infected Suffolk ewe (derived from USA), whereas a rhesus macaque and a chimpanzee exposed to the same inoculum remained healthy 9 years post-exposure1. This inoculum also induced TSE in squirrel monkeys after 4 passages in mice. Other scrapie transmission attempts in macaque failed but had more shorter periods of observation in comparison to the current study. Further, it is possible that there are differences in the zoonotic potential of different scrapie strains.
The most striking observation in our study is the extended incubation period of scrapie in the macaque model, which has several implications. Firstly, our observations constitute experimental evidence in favor of the zoonotic potential of c-scrapie, at least for this isolate that has been extensively studied32,33,34,35,36. The cross-species zoonotic ability of this isolate should be confirmed by performing duplicate intracerebral exposures and assessing the transmissibility by the oral route (a successful transmission of prion strains through the intracerebral route may not necessarily indicate the potential for oral transmission37). However, such confirmatory experiments may require more than one decade, which is hardly compatible with current general management and support of scientific projects; thus this study should be rather considered as a case report.
Secondly, transmission of c-BSE to primates occurred within 8 years post exposure for the lowest doses able to transmit the disease (the survival period after inoculation is inversely proportional to the initial amount of infectious inoculum). The occurrence of scrapie 10 years after exposure to a high dose (25 mg) of scrapie-infected sheep brain suggests that the macaque has a higher species barrier for sheep c-scrapie than c-BSE, although it is notable that previous studies based on in vitro conversion of PrP suggested that BSE and scrapie prions would have a similar conversion potential for human PrP38.
Thirdly, prion diseases typically have longer incubation periods after oral exposure than after intracerebral inoculations: since humans can develop Kuru 47 years after oral exposure39, an incubation time of several decades after oral exposure to scrapie would therefore be expected, leading the disease to occur in older adults, i.e. the peak age for cases considered to be sporadic disease, and making a distinction between scrapie-associated and truly sporadic disease extremely difficult to appreciate.
Fourthly, epidemiologic evidence is necessary to confirm the zoonotic potential of an animal disease suggested by experimental studies. A relatively short incubation period and a peculiar epidemiological situation (e.g., all the first vCJD cases occurring in the country with the most important ongoing c-BSE epizootic) led to a high degree of suspicion that c-BSE was the cause of vCJD. Sporadic CJD are considered spontaneous diseases with an almost stable and constant worldwide prevalence (0.5–2 cases per million inhabitants per year), and previous epidemiological studies were unable to draw a link between sCJD and classical scrapie6,7,40,41, even though external causes were hypothesized to explain the occurrence of some sCJD clusters42,43,44. However, extended incubation periods exceeding several decades would impair the predictive values of epidemiological surveillance for prion diseases, already weakened by a limited prevalence of prion diseases and the multiplicity of isolates gathered under the phenotypes of “scrapie” and “sporadic CJD”.
Fifthly, considering this 10 year-long incubation period, together with both laboratory and epidemiological evidence of decade or longer intervals between infection and clinical onset of disease, no premature conclusions should be drawn from negative transmission studies in cynomolgus macaques with less than a decade of observation, as in the aforementioned historical transmission studies of scrapie to primates1,8,9. Our observations and those of others45,46 to date are unable to provide definitive evidence regarding the zoonotic potential of CWD, atypical/Nor98 scrapie or H-type BSE. The extended incubation period of the scrapie-affected macaque in the current study also underscores the limitations of rodent models expressing human PrP for assessing the zoonotic potential of some prion diseases since their lifespan remains limited to approximately two years21,47,48. This point is illustrated by the fact that the recently reported transmission of scrapie to humanized mice was not associated with clinical signs for up to 750 days and occurred in an extreme minority of mice with only a marginal increase in attack rate upon second passage13. The low attack rate in these studies is certainly linked to the limited lifespan of mice compared to the very long periods of observation necessary to demonstrate the development of scrapie. Alternatively, one could estimate that a successful second passage is the result of strain adaptation to the species barrier, thus poorly relevant of the real zoonotic potential of the original scrapie isolate of sheep origin49. The development of scrapie in this primate after an incubation period compatible with its lifespan complements the study conducted in transgenic (humanized) mice; taken together these studies suggest that some isolates of sheep scrapie can promote misfolding of the human prion protein and that scrapie can develop within the lifespan of some primate species.
In addition to previous studies on scrapie transmission to primate1,8,9 and the recently published study on transgenic humanized mice13, our results constitute new evidence for recommending that the potential risk of scrapie for human health should not be dismissed. Indeed, human PrP transgenic mice and primates are the most relevant models for investigating the human transmission barrier. To what extent such models are informative for measuring the zoonotic potential of an animal TSE under field exposure conditions is unknown. During the past decades, many protective measures have been successfully implemented to protect cattle from the spread of c-BSE, and some of these measures have been extended to sheep and goats to protect from scrapie according to the principle of precaution. Since cases of c-BSE have greatly reduced in number, those protective measures are currently being challenged and relaxed in the absence of other known zoonotic animal prion disease. We recommend that risk managers should be aware of the long term potential risk to human health of at least certain scrapie isolates, notably for lymphotropic strains like the classical scrapie strain used in the current study. Relatively high amounts of infectivity in peripheral lymphoid organs in animals infected with these strains could lead to contamination of food products produced for human consumption. Efforts should also be maintained to further assess the zoonotic potential of other animal prion strains in long-term studies, notably lymphotropic strains with high prevalence like CWD, which is spreading across North America, and atypical/Nor98 scrapie (Nor98)50 that was first detected in the past two decades and now represents approximately half of all reported cases of prion diseases in small ruminants worldwide, including territories previously considered as scrapie free... Even if the prevailing view is that sporadic CJD is due to the spontaneous formation of CJD prions, it remains possible that its apparent sporadic nature may, at least in part, result from our limited capacity to identify an environmental origin.
Singeltary on Scrapie and human transmission way back, see;
MONDAY, NOVEMBER 06, 2017
***> Experimental transfusion of variant CJD-infected blood reveals previously uncharacterised prion disorder in mice and macaque
***> ''On secondary and tertiary transmissions, however, the proportion of PrPres positive animals gradually increased to almost 100%. Recent communications suggest that a similar situation might exist in other models of experimental exposure to prions involving swine32 and cattle33. ''
***> ''Experimental transfusion of variant CJD-infected blood reveals previously uncharacterised prion disorder in mice and macaque''
SATURDAY, DECEMBER 02, 2017
Public health risks from subclinical variant CJD
MONDAY, OCTOBER 02, 2017
Creutzfeldt Jakob Disease United States of America USA and United Kingdom UK Increasing and Zoonotic Pontential From Different Species
THURSDAY, AUGUST 17, 2017
*** Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States revisited 2017
Singeltary et al
U.K.
DEATHS OF DEFINITE AND PROBABLE CJD
† Referral figure for 1990 is from 1 May onwards
* As at 5th March 2018
Summary of vCJD cases
Deaths
Deaths from definite vCJD (confirmed): 123
Deaths from probable vCJD (without neuropathological confirmation): 55
Deaths from probable vCJD (neuropathological confirmation pending): 0
Number of deaths from definite or probable vCJD (as above): 178
Alive
Number of definite/probable vCJD cases still alive: 0
Total number of definite or probable vCJD (dead and alive): 178
1 There are in addition a total of 14 cases of VPSPr (death in 1997(1 case), 2004(1), 2006(1), 2008(3), 2010(1), 2012(4), 2013(1), 2016(1), 2017(1)) not included in the above figures.
2 includes all genetic prion disease, including GSS.
sporadic CJD totals 1939 cases
iatrogenic CJD totals 80 cases
genetic2 CJD totals 191 cases
vCJD totals 178
U.S.A.
PLEASE NOTE, from 1999, where 65 cases of sporadic cjd was recorded, and NO VPSPR or sporadic GSS recorded, to 2016 where 247 cases of sporadic cjd was recorded, where
>>>***PLEASE NOTE***<<<
8 The sporadic cases include 3416 cases of sporadic Creutzfeldt-Jakob disease (sCJD),
***62 cases of Variably Protease-Sensitive Prionopathy (VPSPr) and
***33 cases of sporadic Fatal Insomnia (sFI).
National Prion Disease Pathology Surveillance Center Cases Examined1 (February 16, 2018)
Year Total Referrals2 Prion Disease Sporadic Familial Iatrogenic vCJD
TOTAL 6555 3949(7) 3511(8) 392(9) 12 4
1 Listed based on the year of death or, if not available, on year of referral;
2 Cases with suspected prion disease for which brain tissue was submitted;
3 Disease acquired in the United Kingdom;
4 Disease acquired in the United Kingdom in one case and in Saudi Arabia in the other;
5 Disease possibly acquired in a Middle Eastern or Eastern European country;
6 Includes 33 cases in which the diagnosis is pending, and 20 inconclusive cases;
7 Includes 30 (5 from 2018) cases with type determination pending in which the diagnosis of vCJD has been excluded.
8 The sporadic cases include 3416 cases of sporadic Creutzfeldt-Jakob disease (sCJD), 62 cases of Variably Protease-Sensitive Prionopathy (VPSPr) and 33 cases of sporadic Fatal Insomnia (sFI).
9 Total does not include 237 Familial cases diagnosed by blood test only.
TUESDAY, DECEMBER 12, 2017
Creutzfeldt Jakob Disease CJD National Prion Disease Pathology Surveillance Center Cases Examined to December 14, 2017
***> 8The sporadic cases include 3455 cases of sporadic Creutzfeldt-Jakob disease (sCJD),
62 cases of Variably Protease-Sensitive Prionopathy (VPSPr) and
33 cases of sporadic Fatal Insomnia (sFI).
TUESDAY, JULY 31, 2018
USA CJD TSE Tables of Cases Examined National Prion Disease Pathology Surveillance Center Cases Examined May 1, 2018
http://prionunitusaupdate.blogspot.com/2018/07/usa-cjd-tse-tables-of-cases-examined.html
THURSDAY, OCTOBER 04, 2018
National Prion Disease Pathology Surveillance Center Cases Examined¹ (September 18, 2018)
TUESDAY, NOVEMBER 20, 2018
CDC Eyes of CJD patients show evidence of prions concerns for iatrogenic transmission
MONDAY, DECEMBER 03, 2018
Prion Seeds Distribute throughout the Eyes of Sporadic Creutzfeldt-Jakob Disease Patients
Friday, November 30, 2007
CJD QUESTIONNAIRE USA CWRU AND CJD FOUNDATION
Sunday, August 09, 2009
CJD...Straight talk with...James Ironside...and...Terry Singeltary... 2009
Tuesday, August 18, 2009
BSE-The Untold Story - joe gibbs and singeltary 1999 – 2009
oh what dangerous webs of deceit we weave, when all we do is practice to deceive $$$
HUMAN MAD COW DISEASE nvCJD TEXAS CASE NOT LINKED TO EUROPEAN TRAVEL CDC
Sunday, November 23, 2014
Confirmed Variant Creutzfeldt-Jakob Disease (variant CJD) Case in Texas
Updated: October 7, 2014
CDC and the Texas Department of State Health Services (DSHS) have completed the investigation of the recently reported fourth vCJD case in the United States. It confirmed that the case was in a US citizen born outside the Americas and indicated that the patient's exposure to the BSE/vCJD agent most likely occurred before he moved to the United States; the patient had resided in Kuwait, Russia and Lebanon. The completed investigation did not support the patient's having had extended travel to European countries, including the United Kingdom, or travel to Saudi Arabia. The specific overseas country where this patient’s infection occurred is less clear largely because the investigation did not definitely link him to a country where other known vCJD cases likely had been infected.
MONDAY, AUGUST 14, 2017
Singeltary on Texas Chronic Wasting Disease CWD TSE Prion History
THURSDAY, OCTOBER 22, 2015
Former Ag Secretary Ann Veneman talks women in agriculture and we talk mad cow disease USDA and what really happened to that other mad cow in Texas
spontaneous TSE Prion disease, LOL!
Transmission of scrapie prions to primate after an extended silent incubation period
***Moreover, sporadic disease has never been observed in breeding colonies or primate research laboratories, most notably among hundreds of animals over several decades of study at the National Institutes of Health25, and in nearly twenty older animals continuously housed in our own facility.***
FRIDAY, NOVEMBER 30, 2018
***> The European Union summary report on surveillance for the presence of transmissible spongiform encephalopathies (TSEs) in 2017
DECEMBER 14, 2018, 21 YEARS POST DOD MOM HEIDENHAIN VARIANT CREUTZFELDT JAKOB DISEASE HVCJD DECEMBER 14, 1997, JUST MADE A PROMISE TO MOM, AND YOU DON'T BREAK PROMISES WITH YOUR MOM, NEVER FORGET, AND NEVER LET THEM FORGET...
wasted days and wasted nights...Freddy Fender
TERRY S. SINGELTARY SR., Bacliff, Texas USA 77518 <flounder9@verizon.net>
