Prion seeding activity and infectivity in skin samples from patients with sporadic Creutzfeldt-Jakob disease
Christina D. Orrú1,*, Jue Yuan2,*, Brian S. Appleby2,3,4,*, Baiya Li2,5,*, Yu Li2,6, Dane Winner7, Zerui Wang2,8, Yi-An Zhan2,6, Mark Rodgers2, Jason Rarick2, Robert E. Wyza9, Tripti Joshi9, Gong-Xian Wang6, Mark L. Cohen2, Shulin Zhang2, Bradley R. Groveman1, Robert B. Petersen10, James W. Ironside11, Miguel E. Quiñones-Mateu2,7, Jiri G. Safar2,4, Qingzhong Kong2,3,4,12,†, Byron Caughey1,† and Wen-Quan Zou2,3,4,6,8,12,13,† See all authors and affiliations
Science Translational Medicine 22 Nov 2017: Vol. 9, Issue 417, eaam7785 DOI: 10.1126/scitranslmed.aam7785
Prions in unexpected places
Sporadic Creutzfeldt-Jakob disease (sCJD), the most common human prion disease, can be transmitted via neurosurgical instruments or corneal or dura mater transplants contaminated by infectious prions. Some epidemiological studies have associated sCJD risk with surgeries that involve the skin, but whether the skin of sCJD patients contains prion infectivity is not known. Orrú et al. now report detectable prion seeding activity and infectivity in skin from sCJD patients, although at much lower levels compared to brain tissues from sCJD patients. These data suggest that there may be a potential for iatrogenic sCJD transmission through skin.
Abstract
Sporadic Creutzfeldt-Jakob disease (sCJD), the most common human prion disease, is transmissible through iatrogenic routes due to abundant infectious prions [misfolded forms of the prion protein (PrPSc)] in the central nervous system (CNS). Some epidemiological studies have associated sCJD risk with non-CNS surgeries. We explored the potential prion seeding activity and infectivity of skin from sCJD patients. Autopsy or biopsy skin samples from 38 patients [21 sCJD, 2 variant CJD (vCJD), and 15 non-CJD] were analyzed by Western blotting and real-time quaking-induced conversion (RT-QuIC) for PrPSc. Skin samples from two patients were further examined for prion infectivity by bioassay using two lines of humanized transgenic mice. Western blotting revealed dermal PrPSc in one of five deceased sCJD patients and one of two vCJD patients. However, the more sensitive RT-QuIC assay detected prion seeding activity in skin from all 23 CJD decedents but not in skin from any non-CJD control individuals (with other neurological conditions or other diseases) during blinded testing. 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. Our study demonstrates that the skin of sCJD patients contains both prion seeding activity and infectivity, which raises concerns about the potential for iatrogenic sCJD transmission via skin.
Early preclinical detection of prions in the skin of prion-infected animals
Zerui Wang,#1,2 Matteo Manca,#3 Aaron Foutz,#4 Manuel V. Camacho,#1 Gregory J. Raymond,3 Brent Race,3 Christina D. Orru,3 Jue Yuan,1 Pingping Shen,1,2 Baiya Li,1,5 Yue Lang,1,2 Johnny Dang,1 Alise Adornato,1 Katie Williams,3 Nicholas R. Maurer,1 Pierluigi Gambetti,1 Bin Xu,6 Witold Surewicz,7 Robert B. Petersen,1,8 Xiaoping Dong,9 Brian S. Appleby,1,4,10 Byron Caughey,corresponding author3 Li Cui,corresponding author2 Qingzhong Kong,corresponding author1,4,10,11 and Wen-Quan Zoucorresponding author1,2,4,9,10,11 Author information Article notes Copyright and License information Disclaimer This article has been corrected. See Nat Commun. 2019 February 4; 10: 640. This article has been cited by other articles in PMC. Associated Data Supplementary Materials Data Availability Statement Go to:
Introduction Prion diseases are fatal transmissible neurodegenerative diseases affecting both humans and animals. They include scrapie in sheep and goats, bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in elk, deer, and moose, as well as Creutzfeldt-Jakob disease (CJD), kuru, fatal insomnia, Gerstmann–Sträussler–Scheinker syndrome and variably protease-sensitive prionopathy in humans1. All these diseases are associated with the infectious misfolded form (prion or PrPSc) of a cellular prion protein (PrPC) through a high α-helix structure to high β-sheet conformational transition2,3.
Although PrPC is expressed in virtually all tissues and organs, PrPSc is mainly deposited in the central nervous system and linked to the spongiform degeneration and neuronal loss that are the neuropathological hallmarks of prion diseases. A definitive diagnosis of prion disease has historically mainly depended on examination of brain tissues obtained by biopsy or at autopsy for the presence of prions and neuropathological changes. The recent development of cerebrospinal fluid (CSF)- and nasal brushings-based real-time quaking-induced conversion (RT-QuIC) analysis provides an alternative approach4–6, but to date has only been validated for the diagnosis of clinical-stage prion disease4–9. Moreover, lumbar puncture for CSF sampling is not feasible for all patients due to contraindications and complications in certain conditions. RT-QuIC and serial protein misfolding cyclic amplification (sPMCA) analysis of urine and blood have not been helpful for diagnosing sCJD patients. For instance, PrPSc was detectable in the urine from patients with variant CJD (vCJD, a distinct CJD that originated from exposure to BSE), but, was at much lower rate in sCJD10. Using blood-based sPMCA, PrPSc was detectable in vCJD and in only 1 out of 67 sCJD patients11, consistent with the finding that prion transmission between individuals through blood transfusion has only been reported for vCJD but not sCJD12. There is a need to seek additional readily accessible specimens for the detection of prions in preclinical diagnosis, and for monitoring of disease progression and therapeutic efficiency.
Our recent finding of infectious prions in the skin of patients with sCJD and vCJD raised the possibility that skin PrPSc could be used as a biomarker for early diagnosis and assessment of disease progression13. To test this hypothesis, we examine skin PrPSc in hamsters and humanized transgenic (Tg) mice at different time points after intracerebral prion inoculation using the highly sensitive sPMCA and RT-QuIC assays. We reveal that PrPSc can be detected in the skin of scrapie-infected hamsters at 2 weeks post inoculation (wpi) and human prion-infected humanized Tg mice at 4 wpi by sPMCA as well as skin prion-seeding activity be detected at 3 wpi in hamsters and 20 wpi in Tg mice by RT-QuIC assay. It is worth noting that, compared to the 263K-inoculated hamsters, the mock-inoculated hamsters that has a longer cohabitation period with infected animals exhibit amplified PrPSc by sPMCA in both skin and brain tissues without clinical signs or detectable brain PrPSc by conventional western blotting.
Go to: Results Detection of skin PrPSc by sPMCA in prion-infected hamsters The inoculated animals were killed at 0.4, 1, 2, 3, 4, 7, 10, and 11 weeks post inoculation (wpi) with the 263K scrapie strain, and skin and brain tissues were collected. Clinical signs of scrapie-infected animals appeared from 10 wpi. The deposition of PrPSc and spongiform degeneration became detectable by conventional western blotting, immunohistochemistry, and H&E staining in the brain at 4 wpi and 7 wpi, respectively (Supplementary Figs. 1, 2).
The skin samples from thigh, back, and belly areas of 263K-inoculated hamsters were examined using serial protein misfolding cyclic amplification (sPMCA), a highly sensitive assay for amplification of small amounts of PrPSc14,15. In the thigh skin of scrapie-inoculated animals, the PrPSc was first detected by western blot in the 4th round of sPMCA for 2 wpi samples, in the 3rd round of sPMCA for 7 wpi samples, and in the 2nd round of sPMCA for 10 and 11 wpi samples (Fig. 1a, b, Supplementary Fig. 3), indicating increasing levels of PrPSc over time. The same trend was observed in skin samples from the back and the belly areas of infected animals (Fig. 1b). In contrast, no PrPSc was detected after 8 rounds of sPMCA in the skin of thigh, back and belly from 0.4 to 1 wpi 263K-inoculated hamsters (Supplementary Fig. 4) or 12 wpi-negative control animals intracerebrally inoculated with PBS (Supplementary Fig. 5). Therefore, PrPSc became detectable by sPMCA in the skin of the scrapie-infected animals 5 weeks ahead of brain spongiosis and 8 weeks before the first signs of clinical symptoms.
TUESDAY, MARCH 12, 2019
Early preclinical detection of prions in the skin of prion-infected animals
Nature Reviews Neurology | Review
Advanced tests for early and accurate diagnosis of Creutzfeldt–Jakob disease
Gianluigi Zanusso,1, Salvatore Monaco,1, Maurizio Pocchiari2, & Byron Caughey3, Affiliations Contributions Corresponding author Journal name: Nature Reviews Neurology Year published: (2016) DOI: doi:10.1038/nrneurol.2016.65 Published online 13 May 2016 Abstract Abstract• References• Author information• Supplementary information
Early and accurate diagnosis of Creutzfeldt–Jakob disease (CJD) is a necessary to distinguish this untreatable disease from treatable rapidly progressive dementias, and to prevent iatrogenic transmission. Currently, definitive diagnosis of CJD requires detection of the abnormally folded, CJD-specific form of protease-resistant prion protein (PrPCJD) in brain tissue obtained postmortem or via biopsy; therefore, diagnosis of sporadic CJD in clinical practice is often challenging. Supporting investigations, including MRI, EEG and conventional analyses of cerebrospinal fluid (CSF) biomarkers, are helpful in the diagnostic work-up, but do not allow definitive diagnosis. Recently, novel ultrasensitive seeding assays, based on the amplified detection of PrPCJD, have improved the diagnostic process; for example, real-time quaking-induced conversion (RT-QuIC) is a sensitive method to detect prion-seeding activity in brain homogenate from humans with any subtype of sporadic CJD. RT-QuIC can also be used for in vivo diagnosis of CJD: its diagnostic sensitivity in detecting PrPCJD in CSF samples is 96%, and its specificity is 100%. Recently, we provided evidence that RT-QuIC of olfactory mucosa brushings is a 97% sensitive and 100% specific for sporadic CJD. These assays provide a basis for definitive antemortem diagnosis of prion diseases and, in doing so, improve prospects for reducing the risk of prion transmission. Moreover, they can be used to evaluate outcome measures in therapeutic trials for these as yet untreatable infections.
Subject terms: Diagnostic markers• Prion diseases
Tuesday, May 10, 2016
Accessing transmissibility and diagnostic marker of skin prions 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)
Tuesday, May 10, 2016
Accessing transmissibility and diagnostic marker of skin prions
Tuesday, May 10, 2016 2015
PDA Virus & TSE Safety Forum Meeting Report
>>>Recently transmission of prions from blood of patients with sporadic CJD to humanized mice could be demonstrated.<<<
>>>Further-on, urine samples of a control population (normal and neurological population) showed no signal in the study;
*** however, in samples from patients with sporadic CJD and vCJD, a signal was detected in both patient populations.<<<
Meeting Report: 2015 PDA Virus & TSE Safety Forum
Recent reports suggest that, in addition to meat, bodily fluids such as blood, saliva, feces, and milk may well be risk factors for possible transmission of TSEs to humans. Successful oral transmission among different animal species (interspecies) has been demonstrated. However, species specificity, the “ species barrier,” and the mode of transmission must be taken into account and may explain why cattle, sheep, goats, mink, and mice are successfully orally infected with bovine scrapie-type prion protein (bovPrPSc), whereas the ingestion of bovPrPSc by pigs, poultry, and cervids such as elk and deer fails to cause disease [1]. Humans are also thought to be susceptible to oral infection by bovPrPSc by means of contaminated bovine products (eg, meat pies), and this is believed to be the manner in which the zoonotic disease vCJD originated [4].
Sunday, October 27, 2013
A Kiss of a Prion: New Implications for Oral Transmissibility
Monday, September 19, 2016
Evidence of scrapie transmission to sheep via goat milk
***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;
UPDATE* NOVEMBER 16, 2014
vpspr, sgss, sffi, TSE, an iatrogenic by-product of gss, ffi, familial type prion disease, what it ???
Friday, January 10, 2014
Greetings again Friends, Neighbors, and Colleagues, I would kindly like to follow up on ‘vpspr, sgss, sffi, TSE, an iatrogenic by-product of gss, ffi, familial type prion disease, what it ???’ ran across an old paper from 1984, that some might find interest in, and I will update the link with this old science paper from 1984, a 2010 paper from Japan, and some information on scrapie transmission. The paper from Japan first, then the 1984 paper, and then the scrapie transmission studies.
***The occurrence of contact cases raises the possibility that transmission in families may be effected by an unusually virulent strain of the agent.
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.
85%+ of all human TSE prion, i.e. sporadic CJD, does NOT happen spontaneously, as some would wish you to think. never say never with the TSE Prion disease. ...terry
***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.***
Monday, May 02, 2016
*** Zoonotic Potential of CWD Prions: An Update Prion 2016 Tokyo
SCRAPIE AND CWD ZOONOSIS PRION 2016 CONFERENCE TOKYO
Saturday, April 23, 2016
*** SCRAPIE WS-01: Prion diseases in animals and zoonotic potential 2016
WEDNESDAY, NOVEMBER 20, 2019
Review: Update on Classical and Atypical Scrapie in Sheep and Goats
FRIDAY, NOVEMBER 08, 2019
EFSA Panel on Biological Hazards (BIOHAZ) Update on chronic wasting disease (CWD) III
SUNDAY, APRIL 14, 2019
Estimation of prion infectivity in tissues of cattle infected with atypical BSE by real time-quaking induced conversion assay
WEDNESDAY, APRIL 24, 2019
USDA Announces Atypical Bovine Spongiform Encephalopathy Detection Aug 29, 2018 A Review of Science 2019
WEDNESDAY, JULY 31, 2019
The agent of transmissible mink encephalopathy passaged in sheep is similar to BSE-L
Terry S. Singeltary Sr.