Small critical RNAs in the scrapie agent

Unconventional infectious agents cause transmissible spongiform encephalopathy (TSE) diseases including scrapie and bovine spongiform encephalopathy (BSE) in animals and Creutzfeldt-Jakob disease in humans. The protein only hypothesis claims that the TSE agent is composed solely of the protein called prion (PrP^sc^)^1^. This protein is the misfolded form of a host-encoded cellular protein, PrP^c^ exerting presumably a vital role at the synapse^2^. Even though now widely accepted, the prion concept fails to provide in certain circumstances^3-6^, a satisfying interpretation of the infectious phenomenon. Using the 263K scrapie-hamster model, we conducted a transmission study to search for a putative prion-associated factor indispensable for infectivity. Here we show that innocuous recombinant prion protein (recPrP) was capable, in a reproducible manner, of transmitting scrapie disease when the protein was [beta]–sheet converted in a solution containing PrP^sc^-derived RNA material. Analysis of the PrP-RNA mixture revealed the association of recPrP with two prominent populations of small RNA molecules having an average length of about ~27 and ~55 nucleotides. We conclude that the nature of the TSE agent seems to be composed of a nucleoprotein molecular complex, in which informative RNA molecules of small sizes are associated with the misfolded prion protein (PrP^sc^)

Biology of prions : from the molecular enigma to current risks

Prions are in many ways a real biological enigma, given that the exact nature of the pathogen is still unclear. Bovine Spongiform Encephalopathy (BSE) in Great-Britain, and its related disorder the variant form of Creutzfeldt-Jakob Disease (v-CJD) represent major challenges in terms of animal and human health, given the risks of food-borne contamination, and of secondary man-to-man contamination. Improved knowledge of the biology of these agents has led to major progress over the past few years, notably in the field of diagnosis, decontamination and risk analysis, thereby improving the management of these diseases.Les prions constituent par bien des points une énigme biologique. Notamment, leur nature exacte en tant qu'agent infectieux n'est pas définie précisément. L'Encéphalopathie Spongiforme Bovine (ESB) en Grande-Bretagne, et son corollaire chez l'Homme, la variante de la Maladie de Creutzfeldt-Jakob (v-MCJ), représentent des défis importants en termes de santés animale et humaine, compte tenu des risques de contaminations par voie alimentaire d'une part, et de contaminations secondaires interhumaines d'autre part. La meilleure connaissance de la biologie de ces agents a permis des avancées pratiques importantes ces dernières années, notamment en termes de diagnostic, de décontamination et d'évaluation des risques, permettant ainsi une meilleure gestion de ces maladies

Bimodality, prion aggregates infectivity and prediction of strain phenomenon

21 pagesWe consider a model for the polymerization (fragmentation) process involved in infectious prion self-replication and study both its dynamics and non-zero steady state. We address several issues. Firstly, we give conditions leading to size repartitions of PrPsc aggregates that exhibit bimodal distributions, as indicated by recent experimental studies of prion aggregates distribution. Secondly, we show stability results for this steady state for general coefficients where reduction to a system of differential equations is not possible. We use a duality method based on recent ideas developed for population models. These results underline the potential influence of the amyloid precursor production rate in promoting amyloidogenic diseases. Finally, we numerically investigate the influence of different parameters of the model on PrPsc accumulation kinetics, in the aim to study specific features of prion strains

Diagnostic tests for human and animal prion diseases

The potential existence of clinically silent cases of bovine spongiform encephalopathy (BSE) among cattle, and of humans incubating the new variant Creutzfeldt-Jakob disease (nvCJD) is still a major public health concern. Therefore, the development of screening tests for transmissible subacute spongiform encephalopathies (TSSE) in man and animals remains a priority. In the first part of this paper, we review the main methods used to diagnose generally clinical TSSE, such as brain imaging, electroencephalogram (EEG) analysis, and cerebrospinal fluid (CSF) analysis. In the second part, we present the post-mortem tests used to confirm a TSSE diagnosis, such as inoculation to laboratory animals, histological examination, and identification of abnormal prion protein (PrPres) using biochemical methods. Finally, the third part presents so-called rapid tests (Prionics, Bio-rad, Enfer), validated by the European Commission (EC) for post-slaughter BSE diagnosis in cattle. Now used on a large scale in Europe, these tests have helped assess the extent of the epizooty and eliminate from the food chain animals presenting a risk for human consumption. Since 2002, they have been used for the post-slaughter diagnosis of scrapie in small ruminants. New tests have recently been evaluated by the EC, but it is too soon to predict their role in the field.L'existence potentielle de bovins en phase de latence cliniquement silencieuse d'encéphalopathie spongiforme bovine (ESB) et d'individus en période d'incubation de la nouvelle variante de la maladie de Creutzfeldt-Jakob représente constamment un grand risque pour la santé publique. Par conséquent, le développement de tests de dépistage des encéphalopathies spongiformes subaiguës transmissibles (ESST) humaines et animales constitue toujours une priorité. Dans la première partie de cet article, sont décrites les principales méthodes d'orientation permettant d'aider au diagnostic d'une ESST le plus souvent clinique, comme l'imagerie médicale cérébrale, l'analyse de l'électroencéphalogramme (EEG) et l'examen du liquide céphalo-rachidien. Dans la deuxième partie, sont présentés les tests de confirmation post mortem du diagnostic des ESST, comme l'inoculation à l'animal de laboratoire, l'examen histologique et la recherche de la PrPres par des méthodes biochimiques. La troisième partie est consacrée aux tests dits « rapides » (Prionics, Bio-rad, Enfer), validés en 1999 par la Commission Européenne (CE), pour le diagnostic post mortem de l'ESB à l'abattoir chez les bovins. Utilisés à grande échelle en Europe, ils ont permis de préciser l'étendue réelle de l'épizootie et d'éliminer efficacement de la chaîne alimentaire les animaux présentant un risque pour l'homme. Depuis 2002, ils sont également utilisés pour le diagnostic post mortem des petits ruminants. De nouveaux tests ont été récemment évalués par la CE, mais il est trop tôt pour évaluer la place qu'ils tiendront sur le terrain

Atypical BSE (BASE) Transmitted from Asymptomatic Aging Cattle to a Primate

BACKGROUND: Human variant Creutzfeldt-Jakob Disease (vCJD) results from foodborne transmission of prions from slaughtered cattle with classical Bovine Spongiform Encephalopathy (cBSE). Atypical forms of BSE, which remain mostly asymptomatic in aging cattle, were recently identified at slaughterhouses throughout Europe and North America, raising a question about human susceptibility to these new prion strains. METHODOLOGY/PRINCIPAL FINDINGS: Brain homogenates from cattle with classical BSE and atypical (BASE) infections were inoculated intracerebrally into cynomolgus monkeys (Macacca fascicularis), a non-human primate model previously demonstrated to be susceptible to the original strain of cBSE. The resulting diseases were compared in terms of clinical signs, histology and biochemistry of the abnormal prion protein (PrPres). The single monkey infected with BASE had a shorter survival, and a different clinical evolution, histopathology, and prion protein (PrPres) pattern than was observed for either classical BSE or vCJD-inoculated animals. Also, the biochemical signature of PrPres in the BASE-inoculated animal was found to have a higher proteinase K sensitivity of the octa-repeat region. We found the same biochemical signature in three of four human patients with sporadic CJD and an MM type 2 PrP genotype who lived in the same country as the infected bovine. CONCLUSION/SIGNIFICANCE: Our results point to a possibly higher degree of pathogenicity of BASE than classical BSE in primates and also raise a question about a possible link to one uncommon subset of cases of apparently sporadic CJD. Thus, despite the waning epidemic of classical BSE, the occurrence of atypical strains should temper the urge to relax measures currently in place to protect public health from accidental contamination by BSE-contaminated products

Encephalopathy induced by Alzheimer brain inoculation in a non-human primate.

Alzheimer's disease is characterized by cognitive alterations, cerebral atrophy and neuropathological lesions including neuronal loss, accumulation of misfolded and aggregated β-amyloid peptides (Aβ) and tau proteins. Iatrogenic induction of Aβ is suspected in patients exposed to pituitary-derived hormones, dural grafts, or surgical instruments, presumably contaminated with Aβ. Induction of Aβ and tau lesions has been demonstrated in transgenic mice after contamination with Alzheimer's disease brain homogenates, with very limited functional consequences. Unlike rodents, primates naturally express Aβ or tau under normal conditions and attempts to transmit Alzheimer pathology to primates have been made for decades. However, none of earlier studies performed any detailed functional assessments. For the first time we demonstrate long term memory and learning impairments in a non-human primate (Microcebus murinus) following intracerebral injections with Alzheimer human brain extracts. Animals inoculated with Alzheimer brain homogenates displayed progressive cognitive impairments (clinical tests assessing cognitive and motor functions), modifications of neuronal activity (detected by electroencephalography), widespread and progressive cerebral atrophy (in vivo MRI assessing cerebral volume loss using automated voxel-based analysis), neuronal loss in the hippocampus and entorhinal cortex (post mortem stereology). They displayed parenchymal and vascular Aβ depositions and tau lesions for some of them, in regions close to the inoculation sites. Although these lesions were sparse, they were never detected in control animals. Tau-positive animals had the lowest performances in a memory task and displayed the greatest neuronal loss. Our study is timely and important as it is the first one to highlight neuronal and clinical dysfunction following inoculation of Alzheimer's disease brain homogenates in a primate. Clinical signs in a chronic disease such as Alzheimer take a long time to be detectable. Documentation of clinical deterioration and/or dysfunction following intracerebral inoculations with Alzheimer human brain extracts could lead to important new insights about Alzheimer initiation processes

Hemoglobin mRNA Changes in the Frontal Cortex of Patients with Neurodegenerative Diseases

International audienceBackground: Hemoglobin is the major protein found in erythrocytes, where it acts as an oxygen carrier molecule. In recent years, its expression has been reported also in neurons and glial cells, although its role in brain tissue remains still unknown. Altered hemoglobin expression has been associated with various neurodegenerative disorders. Here, we investigated hemoglobin mRNA levels in brains of patients affected by variant, iatrogenic, and sporadic forms of Creutzfeldt-Jakob disease (vCJD, iCJD, sCJD, respectively) and in different genetic forms of prion diseases (gPrD) in comparison to Alzheimer's disease (AD) subjects and age-matched controls.Methods: Total RNA was obtained from the frontal cortex of vCJD (n = 20), iCJD (n = 11), sCJD (n = 23), gPrD (n = 30), and AD (n = 14) patients and age-matched controls (n = 30). RT-qPCR was performed for hemoglobin transcripts HBB and HBA1/2 using four reference genes for normalization. In addition, expression analysis of the specific erythrocyte marker ALAS2 was performed in order to account for blood contamination of the tissue samples. Hba1/2 and Hbb protein expression was then investigated with immunofluorescence and confocal microscope analysis.Results: We observed a significant up-regulation of HBA1/2 in vCJD brains together with a significant down-regulation of HBB in iCJD. In addition, while in sporadic and genetic forms of prion disease hemoglobin transcripts did not shown any alterations, both chains display a strong down-regulation in AD brains. These results were confirmed also at a protein level.Conclusions: These data indicate distinct hemoglobin transcriptional responses depending on the specific alterations occurring in different neurodegenerative diseases. In particular, the initial site of misfolding event (central nervous system vs. peripheral tissue)—together with specific molecular and conformational features of the pathological agent of the disease—seem to dictate the peculiar hemoglobin dysregulation found in prion and non-prion neurodegenerative disorders.In addition, these results suggest that gene expression of HBB and HBA1/2 in brain tissue is differentially affected by distinct prion and prion-like aggregating protein strains. Validation of these results in more accessible tissues could prompt the development of novel diagnostic tests for neurodegenerative disorders

Evaluation du risque lié à la résistance des prions et développement de techniques adaptées à leur élimination et /ou (inactivation)

PARIS-BIUSJ-Thèses (751052125) / SudocPARIS-BIUSJ-Physique recherche (751052113) / SudocSudocFranceF