RESUMO
Immunohistochemical examination demonstrated widespread granular deposits of alpha-synuclein (alphaSN) in the brains of sheep and goats with natural scrapie, especially in the cornu ammonis and subiculum of the hippocampus; this contrasted with the diffuse and non-granular immunolabelling seen in healthy controls. There was non-regular "co-localization" of PrP(Sc) and alphaSN. The findings resembled those reported in Creutzfeldt-Jakob disease and in experimental prion disease in hamsters and mice. The results suggest that perturbation of alphaSN metabolism plays a role in human and animal prion diseases.
Assuntos
Encéfalo/metabolismo , Doenças das Cabras/metabolismo , Scrapie/metabolismo , alfa-Sinucleína/metabolismo , Animais , Encéfalo/patologia , Feminino , Regulação da Expressão Gênica , Doenças das Cabras/patologia , Cabras , Príons/genética , Príons/metabolismo , Scrapie/patologia , Ovinos , alfa-Sinucleína/genéticaRESUMO
Amphotericin B (AmB) is one of the rare drugs that affect the course of experimental prion diseases and modify the kinetics of abnormal prion protein accumulation in the central nervous system. Therefore, AmB could be used as a pharmacological tool to contribute to our understanding of the pathogenic mechanisms involved in these neurodegenerative disorders.
Assuntos
Anfotericina B/farmacologia , Antibacterianos/farmacologia , Doenças Priônicas/etiologia , Doenças Priônicas/metabolismo , Príons/efeitos dos fármacos , Príons/patogenicidade , Anfotericina B/efeitos adversos , Anfotericina B/química , Animais , Antibacterianos/efeitos adversos , Antibacterianos/química , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Bovinos , Cricetinae , Resistência Microbiana a Medicamentos , Humanos , Camundongos , Estrutura Molecular , Polienos/efeitos adversos , Polienos/química , Polienos/farmacologia , Proteínas PrPSc/efeitos dos fármacos , Príons/metabolismo , OvinosRESUMO
In most experimental models of scrapie and in some naturally infected species, the lymphoreticular system and the spleen in particular play a major role in the pathogenesis of the disease. Previous studies demonstrated scrapie infectivity in peripheral organs from the day of infection up to the terminal stage. The discovery of the abnormal prion protein, PrPres, as a specific molecular hallmark of scrapie should permit enhanced study of scrapie pathogenesis and has some pharmacological applications. In this study, PrPres accumulation was followed day by day in peripheral organs. Four different phases were identified: the circulation of scrapie inoculum, a clearance phase, the peripheral accumulation of PrPres and a plateau phase. This kinetics was then pharmacologically modified (i) by applying the macrophage "suicide" technique to unveil the cellular types involved in scrapie pathogenesis and (ii) with anti-scrapie drugs such as polyene antibiotics, polyanions and Congo red to investigate their mode and site of action.
Assuntos
Anfotericina B/farmacologia , Vermelho Congo/farmacologia , Polímeros/farmacologia , Proteínas PrPSc/metabolismo , Scrapie/tratamento farmacológico , Baço/metabolismo , Anfotericina B/administração & dosagem , Anfotericina B/análogos & derivados , Animais , Ácido Clodrônico/administração & dosagem , Ácido Clodrônico/farmacologia , Vermelho Congo/administração & dosagem , Endopeptidases/metabolismo , Lipossomos , Macrófagos/imunologia , Macrófagos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos SCID , Polieletrólitos , Polímeros/administração & dosagem , Scrapie/metabolismo , Distribuição TecidualRESUMO
The histopathological response of scrapie-infected hamsters treated at the late stage of the infection with an "anti-scrapie" drug, a polyene macrolide antibiotic designated MS-8209, was evaluated in the brain. The results showed that (1) MS-8209 prolonged significantly the incubation time of the experimental disease, (2) MS-8209 delayed the appearance of spongiosis and astrogliosis in the brain, (3) immunodetection of abnormal prion protein and glial fibrillary acidic protein was significantly reduced in the central nervous system. In addition, this report indicates that polyene antibiotics markedly delay the development of the classical brain lesions that result from scrapie infection.
Assuntos
Anfotericina B/análogos & derivados , Antivirais/uso terapêutico , Astrócitos/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Gliose/tratamento farmacológico , Príons/metabolismo , Scrapie/tratamento farmacológico , Anfotericina B/uso terapêutico , Animais , Antibacterianos/uso terapêutico , Astrócitos/patologia , Western Blotting , Encéfalo/metabolismo , Encéfalo/patologia , Cricetinae , Modelos Animais de Doenças , Edema/tratamento farmacológico , Feminino , Proteína Glial Fibrilar Ácida/metabolismo , Gliose/patologia , Imuno-Histoquímica , Mesocricetus , Scrapie/metabolismo , Scrapie/patologiaRESUMO
There is currently no effective therapy available for Creutz-feldt-Jakob disease. However, a limited number of drugs such as polyanions, the amyloid-binding dye Congo red, amphotericin B anthracyclines, dapsone, beta sheet breaker peptides, porphyrines and phtalocyanines have been found to delay the appearance of the clinical signs in experimental prion diseases. Today, the most promising agent would appear to be a less toxic derivative of amphotericin B, MS-8209. Indeed this compound has a wide spectrum of anti-prion activity and is the only molecule capable of prolonging survival time when treatment is performed in the late stages of infection. This result represents an important step forward in therapeutical approaches of prion diseases and justifies the development of new polyene antibiotic derivatives.
Assuntos
Anfotericina B/análogos & derivados , Anfotericina B/uso terapêutico , Antibacterianos/uso terapêutico , Doenças Priônicas/tratamento farmacológico , Animais , Doxorrubicina/análogos & derivados , Doxorrubicina/uso terapêutico , Avaliação Pré-Clínica de Medicamentos , HumanosRESUMO
The observation in 1995 and 1996 of 12 cases of a new variant of Creutzfeldt-Jakob disease (V-CJD) in U.K. suggested a possible relation between this human cases and bovine spongiform encephalopathy (BSE). Recent papers about this topic are reviewed: BSE transmission to macaques, transmission of scrapie with embryo transfer, incidence of maternal transmission, PrP protein released by platelets, diagnostic test by detection of PrP protein in tissues of sheep, epidemiology of BSE, french regulations, identification of cattle in U.K.
Assuntos
Síndrome de Creutzfeldt-Jakob/transmissão , Encefalopatia Espongiforme Bovina/transmissão , Ruminantes , Animais , Bovinos , Síndrome de Creutzfeldt-Jakob/diagnóstico , Síndrome de Creutzfeldt-Jakob/epidemiologia , Encefalopatia Espongiforme Bovina/diagnóstico , Encefalopatia Espongiforme Bovina/epidemiologia , HumanosRESUMO
There is currently no effective therapy available for Creutzfeldt-Jakob disease. However, a limited number of drugs such as polyanions, the amyloid-binding dye Congo red, amphotericin B and anthracyclines have been found to delay the appearance of the clinical signs in experimental prion diseases. Today, the most promising agent appears to be less toxic derivative the amphotericin B, MS-8209. Indeed this compound has a wide spectrum of anti-prion activity and constitute the unique molecule able to prolong survival time when treatment is performed at the late stages of infection. This result represents an important progress in therapeutical approaches of prion diseases and justify the development of new polyene antibiotic derivatives.
Assuntos
Anfotericina B/análogos & derivados , Anfotericina B/uso terapêutico , Antivirais/uso terapêutico , Síndrome de Creutzfeldt-Jakob/tratamento farmacológico , Antibacterianos/uso terapêutico , Antibióticos Antineoplásicos/uso terapêutico , Síndrome de Creutzfeldt-Jakob/patologia , Progressão da Doença , Humanos , Polieletrólitos , Polímeros/uso terapêutico , PrognósticoRESUMO
Creutzfeldt-Jakob disease (CJD) is a rare neurodegenerative illness which belongs to the group of transmissible subacute spongiform encephalopathies (TSSE). Today, no treatment is available for TSSE. The appearance of a new variant of CJD, which affects young people and could be linked to so-called ;mad cow disease', has stimulated researchers to develop new therapies against CJD. A few drugs have already been shown to delay the onset of experimental TSSE. They could contribute to the understanding of the pathogenic mechanisms involved in TSSE and, therefore, could be the basis for therapeutic strategies against CJD.
RESUMO
To test the efficacy of a new amphotericin B derivative, MS-8209, in delaying scrapie, hamsters were infected intracerebrally with the 263K scrapie agent and treated with MS-8209 either early in the course of the disease or continuously. The results show that (i) all treatments lengthened the incubation period of hamster scrapie, (ii) continuous treatment with MS-8209 doubled the length of the incubation period compared with that observed in infected, untreated animals, and (iii) all treatments delayed the accumulation of a proteinase-resistant prion protein and glial fibrillary acidic protein in the brain. These findings suggest that MS-8209 is a powerful tool for investigating the pathogenesis of transmissible subacute spongiform encephalopathies.
Assuntos
Anfotericina B/análogos & derivados , Scrapie/tratamento farmacológico , Anfotericina B/uso terapêutico , Animais , Sistema Nervoso Central/efeitos dos fármacos , Sistema Nervoso Central/metabolismo , Cricetinae , Feminino , Proteína Glial Fibrilar Ácida/biossíntese , Mesocricetus , Proteínas PrPSc/biossíntese , Scrapie/metabolismoRESUMO
Transmissible subacute spongiform encephalopathies (TSSE) are neurodegenerative diseases characterized by the presence of a modified, partially proteinase-resistant host protein, PrPSc, which accumulates in the brains of infected individuals. Recently it has been reported that amphotericin B (AmB) treatment of hamsters infected with scrapie strain 263K prolongs the incubation period of the disease, and dissociates in vivo replication of the scrapie agent from PrPSc accumulation. We report here on data obtained after treatment with AmB and one of its derivatives, MS-8209, in experimental scrapie of mouse and hamster. Treatment was carried out by the intraperitoneal route 6 days per week, at three different dosages initiated at the time of infection. Two regimens were used: during the early time of infection or throughout the experimental infection. Results indicate that MS-8209 was as efficient as AmB in prolonging the incubation time and decreasing PrPSc accumulation in the hamster scrapie model. A dose-dependent response was observed in mice treated early after experimental infection. At a dose of 2.5 mg/kg, MS-8209 significantly prolonged the incubation period (by 11.9%). In long-term treatment of mice, MS-8209 and AmB markedly reduced PrPSc levels in the preclinical stage of the disease. These data demonstrate that the effect of AmB is not restricted to one model (hamster-263K). This regimen leads to an inversion of the PrPSc to proteinase-sensitive protein (PrPSens) ratio, suggesting PrPSens (presumably cellular PrPC) accumulation occurs before its conversion into PrPSc. As it has been shown that AmB does not modify the infectivity titre, we conclude that the drugs could act by inhibiting either the interaction of the scrapie agent with PrPSens during the early times of infection or the conversion of PrPSens into PrPSc.
Assuntos
Anfotericina B/análogos & derivados , Anfotericina B/uso terapêutico , Antivirais/uso terapêutico , Doenças Priônicas/tratamento farmacológico , Príons/biossíntese , Príons/metabolismo , Replicação Viral/efeitos dos fármacos , Animais , Cricetinae , Camundongos , Camundongos Endogâmicos C57BL , Proteínas PrPSc , Príons/efeitos dos fármacos , Príons/isolamento & purificação , Fatores de TempoRESUMO
Mice were infected intracerebrally with the bovine spongiform encephalopathy (BSE) or the scrapie agent and treated during 8 weeks postinfection to test the protective effect of a new amphotericin B (AmB) derivative, MS-8209, in experimental transmissible spongiform encephalopathies. The results show that (i) the treatment prolonged the incubation period of both BSE-infected and scrapie-infected mice, (ii) MS-8209 and AmB were much more efficient in delaying the onset of scrapie than that of BSE, and (iii) a delay in Prp-res (proteinase K-resistant prion protein) and GFAP (glial fibrillary acidic protein) accumulation was observed in the brains of scrapie-infected mice, but was not significant in BSE-infected mice. The analysis of the molecular and clinical results strongly suggests a common mechanism of action of this category of drugs on the different transmissible spongiform encephalopathy strains. This could be due to an interaction with the PrP transconformation process leading to the formation of PrP-res.
Assuntos
Anfotericina B/análogos & derivados , Antivirais/uso terapêutico , Encefalopatia Espongiforme Bovina/tratamento farmacológico , Scrapie/tratamento farmacológico , Anfotericina B/uso terapêutico , Animais , Antibacterianos , Bovinos , Relação Dose-Resposta a Droga , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Polienos , Proteínas PrPSc/metabolismoRESUMO
Amphotericin B derivatives, such as MS-8209, have been evaluated as a therapeutic approach to human immunodeficiency virus (HIV) infection. We show that MS-8209, like amphotericin B, increases tumor necrosis factor alpha (TNF-alpha) mRNA expression and TNF-alpha production and consequently HIV replication in human macrophages. These effects confirm the pharmacological risk associated with the administration of amphotericin B or its derivatives to HIV-infected patients.
Assuntos
Anfotericina B/análogos & derivados , Fármacos Anti-HIV/farmacologia , HIV/efeitos dos fármacos , Macrófagos/virologia , Fator de Necrose Tumoral alfa/biossíntese , Anfotericina B/farmacologia , HIV/fisiologia , Humanos , Macrófagos/metabolismo , RNA Mensageiro/biossíntese , Fatores de Risco , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Replicação Viral/efeitos dos fármacosRESUMO
The mode and the site of action of the major antiscrapie drugs have been studied by investigating their effects on the abnormal protease-resistant isoform of PrP (PrPres) and on its accumulation in mouse spleen. Day-by-day PrPres accumulation in the spleen and in other peripheral organs was first monitored to describe the early steps of scrapie pathogenesis. Three phases were identified: the detection of scrapie inoculum on the day of scrapie infection, a clearance phase, and then the peripheral accumulation of PrPres. In a second step, the effects of the polyene antibiotic MS-8209, the polyanion dextran sulfate 500 (DS500), and Congo red were assessed on these phases, after the drugs were coincubated with scrapie inoculum. Highly different mechanisms and sites of action were apparent. MS-8209 had a weak effect on the accumulation of PrPres in spleen, suggesting another site of intervention for this drug. DS500 delayed the beginning of the clearance phase but then blocked PrPres synthesis for a long period of time, probably because of its immunological effects on the spleen. Surprisingly, Congo red suppressed the clearance phase of scrapie inoculum and then increased transiently accumulation of PrPres in spleen. We showed in vitro that this effect was related to a direct enhancement of the protease resistance of PrPres by the drug.
Assuntos
Anfotericina B/análogos & derivados , Vermelho Congo/farmacologia , Sulfato de Dextrana/farmacologia , Proteínas PrPSc/metabolismo , Scrapie/metabolismo , Baço/efeitos dos fármacos , Anfotericina B/administração & dosagem , Anfotericina B/metabolismo , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Encéfalo/patologia , Vermelho Congo/administração & dosagem , Vermelho Congo/metabolismo , Vermelho Congo/uso terapêutico , Cricetinae , Sulfato de Dextrana/administração & dosagem , Sulfato de Dextrana/metabolismo , Sulfato de Dextrana/uso terapêutico , Relação Dose-Resposta a Droga , Endopeptidase K/metabolismo , Feminino , Injeções Intraperitoneais , Cinética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos SCID , Especificidade de Órgãos , Proteínas PrPSc/administração & dosagem , Proteínas PrPSc/antagonistas & inibidores , Proteínas PrPSc/isolamento & purificação , Ligação Proteica , Isoformas de Proteínas/administração & dosagem , Isoformas de Proteínas/antagonistas & inibidores , Isoformas de Proteínas/isolamento & purificação , Isoformas de Proteínas/metabolismo , Scrapie/tratamento farmacológico , Scrapie/patologia , Baço/química , Baço/metabolismo , Baço/patologia , Fatores de TempoRESUMO
The development of transmissible spongiform encephalopathies in experimental models depends on two major factors: the intracerebral accumulation of an abnormal, protease-resistant isoform of PrP (PrPres), which is a host protein mainly expressed in neurons; and the existence of different strains of agent. In order to make a distinction between pathogenic mechanisms depending upon the accumulation of host-derived PrPres and the strain-specific effects, we quantified and compared the sequence of molecular [PrPres and glial fibrillary acidic protein (GFAP) accumulation] and pathological events in the brains of syngeneic mice throughout the course of infection with two different strains of agent. The bovine spongiform encephalopathy (BSE) agent exhibits properties different from any known scrapie source and has been studied in comparison with a classical scrapie strain. Convergent kinetic data in both models confirmed the cause-effect relationship between PrPres and pathological changes and showed that PrPres accumulation is directly responsible for astrocyte activation in vivo. Moreover, we observed a threshold level of PrPres for this effect on astroglial cells. However, despite similar infectivity titres, the BSE model produced less PrPres than scrapie, and the relative importance of gliosis was higher. The comparison of the molecular and pathological features after intracerebral or intraperitoneal inoculation also revealed differences between the models. Therefore, the mechanisms leading to the targeting and the fine regulation of the molecular events seem to be independent of the host PrP and to depend upon the agent. The possible involvement of a regulatory molecule accounting for these specificities has to be considered.
Assuntos
Encefalopatia Espongiforme Bovina/etiologia , Proteínas PrPSc/patogenicidade , Scrapie/etiologia , Animais , Astrócitos/metabolismo , Bovinos , Modelos Animais de Doenças , Encefalopatia Espongiforme Bovina/patologia , Encefalopatia Espongiforme Bovina/fisiopatologia , Proteína Glial Fibrilar Ácida/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Scrapie/patologia , Scrapie/fisiopatologia , Ovinos , Fatores de TempoRESUMO
Prion related disorders are associated with the accumulation of a misfolded isoform (PrPsc) of the host-encoded prion protein, PrP. There is strong evidence for the involvement of unidentified co-factors in the PrP to PrPsc conversion process. In this study, we show alpha-synuclein-immunoreactive deposits in the central nervous system of various prion diseases (sporadic, iatrogenic and new variant Creutzfeldt-Jakob diseases, and experimental scrapie of hamsters). alpha-Synuclein accumulated close to PrPsc deposits but we did not observe strict colocalization of prion protein and alpha-synuclein immunoreactivities particularly in PrPsc plaques. alpha-Synuclein is thought to be a key player in some neurodegenerative disorders, is able to interact with amyloid structures and has known chaperone-like activities. Our results, in various prion diseases, suggest a role for alpha-synuclein in regulating PrPsc formation.
Assuntos
Sistema Nervoso Central/metabolismo , Síndrome de Creutzfeldt-Jakob/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Neurópilo/metabolismo , Scrapie/metabolismo , Animais , Sistema Nervoso Central/patologia , Sistema Nervoso Central/fisiopatologia , Síndrome de Creutzfeldt-Jakob/patologia , Síndrome de Creutzfeldt-Jakob/fisiopatologia , Cricetinae , Humanos , Imuno-Histoquímica , Corpos de Inclusão/metabolismo , Neurônios/patologia , Neurópilo/patologia , Proteínas PrPSc/metabolismo , Príons/metabolismo , Scrapie/patologia , Scrapie/fisiopatologia , Sinucleínas , alfa-SinucleínaRESUMO
Amphotericin B (AmB) is one of the few drugs able to prolong survival times in experimental scrapie and delays the accumulation of PrPres, a specific marker of this disease in the brain in vivo. Previous reports showed that the AmB effect is observed only if the drug is administered around the time of infection. In the present study, intracerebrally infected mice were treated with AmB or one of its derivatives, MS-8209, between 80 and 140 days postinoculation. We observed an increased incubation time and a delay in PrPres accumulation and glial fibrillary acidic protein gene expression. Treatment starting at 80 days postinoculation was as efficient as long-term treatment starting the day of inoculation. Our results indicate that polyene antibiotics may interfere, throughout the course of the experimental disease, with the propagation of the scrapie agent.
Assuntos
Anfotericina B/análogos & derivados , Polienos/farmacologia , Scrapie/tratamento farmacológico , Sequência de Aminoácidos , Anfotericina B/farmacologia , Animais , Antibacterianos/farmacologia , Esquema de Medicação , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Proteínas PrPSc , Coelhos , Scrapie/fisiopatologia , Fatores de TempoRESUMO
Using the severe combined immunodeficiency (SCID) mouse model, we investigated the requirement of the immune system for the development of scrapie after peripheral inoculation. A total of 33% of SCID mice, all but one immunologically reconstituted SCID mice (93%), and all CB17 control mice developed the disease. PrPres was detectable in the brains of all diseased animals and in the spleens of reconstituted SCID and CB17 control mice but not of the diseased non-immunologically reconstituted SCID mice. The immune system appears to be a primary target in the pathogenesis of scrapie, but direct spread to the central nervous system from the peritoneum via visceral nerve fibers can probably also occur.
Assuntos
Proteínas PrPSc/imunologia , Scrapie/imunologia , Animais , Encéfalo/imunologia , Sistema Imunitário , Camundongos , Camundongos Endogâmicos C57BL , Camundongos SCID , Proteínas PrPSc/biossíntese , Baço/imunologiaRESUMO
Amphotericin B (AmB) has been shown to delay hamster scrapie. Infectivity studies have been performed previously using AmB in order to understand the relationship between the accumulation of an abnormal isoform (PrPres) of the prion protein and 263K scrapie agent replication in the brain. The first study reported that AmB had no effect upon agent replication, although it delayed the development of both clinical signs and PrPres accumulation. However, subsequent experiments using the same model showed a significant effect both on agent replication and PrPres accumulation early in infection. This fundamental discrepancy was assumed to be linked to differences in experimental protocols. In order to unravel the issue, a new experiment has been performed encompassing different protocols and using an AmB derivative, MS-8209, that can be used at higher doses because of its lower toxicity. The findings of this study exclude the suspected differences in the protocols as the reason for previous conflicting results, and suggest strongly that these discrepancies were due to a low dose of AmB causing a 'threshold effect'. Overall, this study indicates that, in this model, PrPres cannot be dissociated from infectivity by polyene antibiotics.
Assuntos
Anfotericina B/análogos & derivados , Antivirais/farmacologia , Proteínas PrPSc/efeitos dos fármacos , Anfotericina B/farmacologia , Animais , Cricetinae , Relação Dose-Resposta a Droga , Feminino , Mesocricetus , Proteínas PrPSc/metabolismoRESUMO
The polyene antibiotic MS-8209 is currently one of the most effective drugs in the treatment of experimental scrapie. However, its mechanism of action and its site of intervention in the pathogenetical process of scrapie infection are largely unknown. It has been shown previously that the infection of immunodeficient SCID mice by the peripheral route provides a reliable model for direct scrapie neuroinvasion, bypassing the lymphoreticular system. Indeed, a proportion of SCID mice develop scrapie after a similar time to immunocompetent mice, despite their severe immune impairment. This model is now used to clarify the targets of MS-8209. In SCID mice, MS-8209 treatment protected against infection but did not prolong survival time. In SCID mice immunologically reconstituted prior to inoculation, the drug delayed the disease without an effect on the attack rate. These findings strongly suggest that MS-8209 acts by hampering the first step of the neuroinvasion process, i.e. the uptake of the infectious agent by peripheral nerve endings. The mechanism leading to the inhibition of agent propagation to nervous cells is discussed with regard to the properties of polyene antibiotics.
Assuntos
Anfotericina B/análogos & derivados , Antivirais/uso terapêutico , Príons/efeitos dos fármacos , Scrapie/prevenção & controle , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico , Animais , Antivirais/farmacologia , Camundongos , Camundongos SCID , Sistema Nervoso/virologiaRESUMO
Based on in vitro observations in scrapie-infected neuroblastoma cells, quinacrine has recently been proposed as a treatment for Creutzfeldt-Jakob disease (CJD), including a new variant CJD which is linked to contamination of food by the bovine spongiform encephalopathy (BSE) agent. The present study investigated possible mechanisms of action of quinacrine on prions. The ability of quinacrine to interact with and to reduce the protease resistance of PrP peptide aggregates and PrPres of human and animal origin were analyzed, together with its ability to inhibit the in vitro conversion of the normal prion protein (PrPc) to the abnormal form (PrPres). Furthermore, the efficiencies of quinacrine and chlorpromazine, another tricyclic compound, were examined in different in vitro models and in an experimental murine model of BSE. Quinacrine efficiently hampered de novo generation of fibrillogenic prion protein and PrPres accumulation in ScN2a cells. However, it was unable to affect the protease resistance of preexisting PrP fibrils and PrPres from brain homogenates, and a "curing" effect was obtained in ScGT1 cells only after lengthy treatment. In vivo, no detectable effect was observed in the animal model used, consistent with other recent studies and preliminary observations in humans. Despite its ability to cross the blood-brain barrier, the use of quinacrine for the treatment of CJD is questionable, at least as a monotherapy. The multistep experimental approach employed here could be used to test new therapeutic regimes before their use in human trials.