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1.
Nature ; 538(7625): 344-349, 2016 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-27602946

RESUMO

Antimalarial drugs have thus far been chiefly derived from two sources-natural products and synthetic drug-like compounds. Here we investigate whether antimalarial agents with novel mechanisms of action could be discovered using a diverse collection of synthetic compounds that have three-dimensional features reminiscent of natural products and are underrepresented in typical screening collections. We report the identification of such compounds with both previously reported and undescribed mechanisms of action, including a series of bicyclic azetidines that inhibit a new antimalarial target, phenylalanyl-tRNA synthetase. These molecules are curative in mice at a single, low dose and show activity against all parasite life stages in multiple in vivo efficacy models. Our findings identify bicyclic azetidines with the potential to both cure and prevent transmission of the disease as well as protect at-risk populations with a single oral dose, highlighting the strength of diversity-oriented synthesis in revealing promising therapeutic targets.


Assuntos
Antimaláricos/síntese química , Antimaláricos/farmacologia , Azetidinas/uso terapêutico , Descoberta de Drogas , Estágios do Ciclo de Vida/efeitos dos fármacos , Malária Falciparum/tratamento farmacológico , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/crescimento & desenvolvimento , Animais , Antimaláricos/administração & dosagem , Antimaláricos/uso terapêutico , Compostos Azabicíclicos/administração & dosagem , Compostos Azabicíclicos/síntese química , Compostos Azabicíclicos/farmacologia , Compostos Azabicíclicos/uso terapêutico , Azetidinas/administração & dosagem , Azetidinas/efeitos adversos , Azetidinas/farmacologia , Citosol/enzimologia , Modelos Animais de Doenças , Feminino , Fígado/efeitos dos fármacos , Fígado/parasitologia , Macaca mulatta/parasitologia , Malária Falciparum/prevenção & controle , Malária Falciparum/transmissão , Masculino , Camundongos , Fenilalanina-tRNA Ligase/antagonistas & inibidores , Compostos de Fenilureia/administração & dosagem , Compostos de Fenilureia/síntese química , Compostos de Fenilureia/farmacologia , Compostos de Fenilureia/uso terapêutico , Plasmodium falciparum/citologia , Plasmodium falciparum/enzimologia , Segurança
2.
Antimicrob Agents Chemother ; 58(8): 4737-44, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24913163

RESUMO

Hematotoxicity in individuals genetically deficient in glucose-6-phosphate dehydrogenase (G6PD) activity is the major limitation of primaquine (PQ), the only antimalarial drug in clinical use for treatment of relapsing Plasmodium vivax malaria. PQ is currently clinically used in its racemic form. A scalable procedure was developed to resolve racemic PQ, thus providing pure enantiomers for the first time for detailed preclinical evaluation and potentially for clinical use. These enantiomers were compared for antiparasitic activity using several mouse models and also for general and hematological toxicities in mice and dogs. (+)-(S)-PQ showed better suppressive and causal prophylactic activity than (-)-(R)-PQ in mice infected with Plasmodium berghei. Similarly, (+)-(S)-PQ was a more potent suppressive agent than (-)-(R)-PQ in a mouse model of Pneumocystis carinii pneumonia. However, at higher doses, (+)-(S)-PQ also showed more systemic toxicity for mice. In beagle dogs, (+)-(S)-PQ caused more methemoglobinemia and was toxic at 5 mg/kg of body weight/day given orally for 3 days, while (-)-(R)-PQ was well tolerated. In a novel mouse model of hemolytic anemia associated with human G6PD deficiency, it was also demonstrated that (-)-(R)-PQ was less hemolytic than (+)-(S)-PQ for the G6PD-deficient human red cells engrafted in the NOD-SCID mice. All these data suggest that while (+)-(S)-PQ shows greater potency in terms of antiparasitic efficacy in rodents, it is also more hematotoxic than (-)-(R)-PQ in mice and dogs. Activity and toxicity differences of PQ enantiomers in different species can be attributed to their different pharmacokinetic and metabolic profiles. Taken together, these studies suggest that (-)-(R)-PQ may have a better safety margin than the racemate in human.


Assuntos
Antimaláricos/farmacocinética , Hemólise/efeitos dos fármacos , Malária/tratamento farmacológico , Pneumonia por Pneumocystis/tratamento farmacológico , Primaquina/farmacocinética , Animais , Antimaláricos/isolamento & purificação , Antimaláricos/toxicidade , Cães , Transfusão de Eritrócitos , Eritrócitos/efeitos dos fármacos , Eritrócitos/parasitologia , Feminino , Deficiência de Glucosefosfato Desidrogenase/metabolismo , Humanos , Dose Letal Mediana , Malária/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos ICR , Camundongos Endogâmicos NOD , Camundongos SCID , Plasmodium berghei/efeitos dos fármacos , Plasmodium berghei/fisiologia , Pneumocystis carinii/efeitos dos fármacos , Pneumocystis carinii/fisiologia , Pneumonia por Pneumocystis/microbiologia , Primaquina/isolamento & purificação , Primaquina/toxicidade , Estereoisomerismo , Transplante Heterólogo
3.
Toxicol Pathol ; 42(8): 1267-74, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24499803

RESUMO

Vascular endothelial growth factor (VEGF) receptor tyrosine kinase (RTK) inhibitors are reported to cause reversible mucosal hyperplasia (adenosis) in the duodenum of rats; however, the pathogenesis is not fully elucidated. Using lenvatinib, a VEGF RTK inhibitor, we characterized the histologic time course of this duodenal change in rats. At 4 weeks, there was degeneration and necrosis of Brunner's gland epithelium accompanied by neutrophil infiltration around the affected glands. At 13 weeks, the inflammation was more extensive, and Brunner's gland epithelium was attenuated and flattened and was accompanied by reactive hyperplasia of duodenal epithelium. At 26 weeks, the changes became more severe and chronic and characterized by marked cystic dilation, which extended to the external muscular layer. These dilated glands exhibited morphological characteristics of duodenal crypt epithelium, suggestive of replacement of disappeared Brunner's glands by regenerative duodenal crypt epithelial cells. Similar changes were not present in similar time course studies in dog and monkey studies, suggesting that this is a rodent- or species-specific change. Based on the temporal progression of Brunner's gland lesion, we identify degeneration and necrosis of the Brunner's glands as the primary change leading to inflammation, cystic dilatation, and regeneration with cells that are morphologically suggestive of duodenal crypt epithelium.


Assuntos
Glândulas Duodenais/efeitos dos fármacos , Duodenopatias/induzido quimicamente , Compostos de Fenilureia/toxicidade , Quinolinas/toxicidade , Receptores Proteína Tirosina Quinases/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Animais , Glândulas Duodenais/citologia , Glândulas Duodenais/patologia , Duodenopatias/patologia , Feminino , Hiperplasia/induzido quimicamente , Hiperplasia/patologia , Inflamação/induzido quimicamente , Inflamação/patologia , Masculino , Compostos de Fenilureia/administração & dosagem , Quinolinas/administração & dosagem , Ratos , Ratos Sprague-Dawley
4.
Lancet Infect Dis ; 20(8): 964-975, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32275867

RESUMO

BACKGROUND: (+)-SJ000557733 (SJ733) is a novel, orally bioavailable inhibitor of Plasmodium falciparum ATP4. In this first-in-human and induced blood-stage malaria phase 1a/b trial, we investigated the safety, tolerability, pharmacokinetics, and antimalarial activity of SJ733 in humans. METHODS: The phase 1a was a single-centre, dose-escalation, first-in-human study of SJ733 allowing modifications to dose increments and dose-cohort size on the basis of safety and pharmacokinetic results. The phase 1a took place at St Jude Children's Research Hospital and at the University of Tennessee Clinical Research Center (Memphis, TN, USA). Enrolment in more than one non-consecutive dose cohort was allowed with at least 14 days required between doses. Participants were fasted in seven dose cohorts and fed in one 600 mg dose cohort. Single ascending doses of SJ733 (75, 150, 300, 600, 900, or 1200 mg) were administered to participants, who were followed up for 14 days after SJ733 dosing. Phase 1a primary endpoints were safety, tolerability, and pharmacokinetics of SJ733, and identification of an SJ733 dose to test in the induced blood-stage malaria model. The phase 1b was a single-centre, open-label, volunteer infection study using the induced blood-stage malaria model in which fasted participants were intravenously infected with blood-stage P falciparum and subsequently treated with a single dose of SJ733. Phase 1b took place at Q-Pharm (Herston, QLD, Australia) and was initiated only after phase 1a showed that exposure exceeding the threshold minimum exposure could be safely achieved in humans. Participants were inoculated on day 0 with P falciparum-infected human erythrocytes (around 2800 parasites in the 150 mg dose cohort and around 2300 parasites in the 600 mg dose cohort), and parasitaemia was monitored before malaria inoculation, after inoculation, immediately before SJ733 dosing, and then post-dose. Participants were treated with SJ733 within 24 h of reaching 5000 parasites per mL or at a clinical score higher than 6. Phase 1b primary endpoints were calculation of a parasite reduction ratio (PRR48) and parasite clearance half-life, and safety and tolerability of SJ733 (incidence, severity, and drug-relatedness of adverse events). In both phases of the trial, SJ733 hydrochloride salt was formulated as a powder blend in capsules containing 75 mg or 300 mg for oral administration. Healthy men and women (of non-childbearing potential) aged 18-55 years were eligible for both studies. Both studies are registered with ClinicalTrials.gov (NCT02661373 for the phase 1a and NCT02867059 for the phase 1b). FINDINGS: In the phase 1a, 23 healthy participants were enrolled and received one to three non-consecutive doses of SJ733 between March 14 and Dec 7, 2016. SJ733 was safe and well tolerated at all doses and in fasted and fed conditions. 119 adverse events were recorded: 54 (45%) were unrelated, 63 (53%) unlikely to be related, and two (2%) possibly related to SJ733. In the phase 1b, 17 malaria-naive, healthy participants were enrolled. Seven participants in the 150 mg dose cohort were inoculated and dosed with SJ733. Eight participants in the 600 mg dose cohort were inoculated, but two participants could not be dosed with SJ733. Two additional participants were subsequently inoculated and dosed with SJ733. SJ733 exposure increased proportional to the dose through to the 600 mg dose, then was saturable at higher doses. Fasted participants receiving 600 mg exceeded the target area under the concentration curve extrapolated to infinity (AUC0-∞) of 13 000 µg × h/L (median AUC0-∞ 24 283 [IQR 16 135-31 311] µg × h/L, median terminal half-life 17·4 h [IQR 16·1-24·0], and median timepoint at which peak plasma concentration is reached 1·0 h [0·6-1·3]), and this dose was tested in the phase 1b. All 15 participants dosed with SJ733 had at least one adverse event. Of the 172 adverse events recorded, 128 (74%) were mild. The only adverse event attributed to SJ733 was mild bilateral foot paraesthesia that lasted 3·75 h and resolved spontaneously. The most common adverse events were related to malaria. Based on parasite clearance half-life, the derived log10PRR48 and corresponding parasite clearance half-lives were 2·2 (95% CI 2·0-2·5) and 6·47 h (95% CI 5·88-7·18) for 150 mg, and 4·1 (3·7-4·4) and 3·56 h (3·29-3·88) for 600 mg. INTERPRETATION: The favourable pharmacokinetic, tolerability, and safety profile of SJ733, and rapid antiparasitic effect support its development as a fast-acting component of combination antimalarial therapy. FUNDING: Global Health Innovative Technology Fund, Medicines for Malaria Venture, and the American Lebanese Syrian Associated Charities.


Assuntos
Antimaláricos/uso terapêutico , Compostos Heterocíclicos de 4 ou mais Anéis/uso terapêutico , Isoquinolinas/uso terapêutico , Malária Falciparum/tratamento farmacológico , Malária Falciparum/parasitologia , Plasmodium falciparum/efeitos dos fármacos , Inibidores da Bomba de Prótons/uso terapêutico , Adulto , Antimaláricos/administração & dosagem , Antimaláricos/efeitos adversos , Antimaláricos/farmacocinética , Estudos de Casos e Controles , Eritrócitos/efeitos dos fármacos , Eritrócitos/parasitologia , Feminino , ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Compostos Heterocíclicos de 4 ou mais Anéis/administração & dosagem , Compostos Heterocíclicos de 4 ou mais Anéis/efeitos adversos , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Humanos , Isoquinolinas/administração & dosagem , Isoquinolinas/efeitos adversos , Isoquinolinas/farmacocinética , Estágios do Ciclo de Vida/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Plasmodium falciparum/crescimento & desenvolvimento , Plasmodium falciparum/metabolismo , Inibidores da Bomba de Prótons/administração & dosagem , Inibidores da Bomba de Prótons/efeitos adversos , Inibidores da Bomba de Prótons/farmacocinética , Resultado do Tratamento , Adulto Jovem
5.
Exp Biol Med (Maywood) ; 229(10): 1061-71, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15522843

RESUMO

Staphylococcal enterotoxin (SE) B causes serious gastrointestinal illness, and intoxication with this exotoxin can lead to lethal toxic shock syndrome. In order to overcome significant shortcomings of current rodent and nonhuman primate models, we developed a piglet model of lethal SEB intoxication. Fourteen-day-old Yorkshire piglets were given intravenous SEB, observed clinically, and sacrificed at 4, 6, 24, 48, 72, or 96 hrs posttreatment. Clinical signs were biphasic with pyrexia, vomiting, and diarrhea within 4 hrs, followed by terminal hypotension and shock by 96 hrs. Mild lymphoid lesions were identified as early as 24 hrs, with severe lymphadenopathy, splenomegaly, and prominent Peyer's patches found by 72 hrs. Widespread edema-most prominent in the mesentery, between loops of spiral colon, and in retroperitoneal connective tissue-was found in animals at 72 hrs. Additional histologic changes included perivascular aggregates of large lymphocytes variably present in the lung and brain, circulating lymphoblasts, and lymphocytic portal hepatitis. Preliminary molecular investigation using gene array has uncovered several gene profile changes that may have implications in the pathophysiology leading to irreversible shock. Five genes were selected for further study, and all showed increased mRNA levels subsequent to SEB exposure. The use of this piglet model will continue to elucidate the pathogenesis of SEB intoxication and facilitate the testing of new therapeutic regimens that may better correlate with human lesions.


Assuntos
Morte , Modelos Animais de Doenças , Enterotoxinas/toxicidade , Choque Séptico/patologia , Animais , Pressão Sanguínea , Encéfalo/patologia , Edema/patologia , Enterotoxinas/administração & dosagem , Feminino , Injeções Intravenosas , Intestino Grosso/patologia , Fígado/patologia , Pulmão/patologia , Linfonodos/patologia , Masculino , Choque Séptico/mortalidade , Suínos , Síndrome , Temperatura , Fatores de Tempo
6.
Comp Med ; 52(6): 513-7, 2002 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12540163

RESUMO

BACKGROUND AND PURPOSE: Use of high-energy near-infrared lasers is becoming more prevalent in today's industries, such as technology, medicine, and military operations. Despite wide-range use of these lasers, threshold, median effective dose (ED50), and the mechanism of laser-tissue interaction are not well defined at the 1,318-nm wavelength for human corneal exposures. The goals of the study reported here were to establish the ED50 for single-pulse, 1,318-nm laser exposures on the Dutch Belted rabbit cornea and to characterize microscopic changes. Results of this study were then compared with those of previous corneal studies. METHODS: A neodymium:yttrium aluminum garnet (Nd:YAG) laser was used to deliver single 1,318-nm wavelength pulses to the corneas of 10 female Dutch Belted rabbits (Oryctolagus cuniculus). Single pulses of 0.5-ms duration and radiant energy ranging from 116 to 2,250 J/cm2 irradiated the exposure sites. Sites were clinically evaluated for presence of a lesion at one hour and 24-h after exposure. Results of the 24-h evaluation were used to determine the (ED50). Corneas were subsequently collected at the 24-h endpoint for microscopic evaluation. RESULTS: The ED50 for 1,318-nm exposures to the rabbit cornea was determined to be 382 J/cm2, as measured at the 1/e2 (0.865 times that of the peak power per unit area). At each exposure site, there was a small (< 1 mm in diameter), white, circular, well demarcated corneal lesion characterized histologically by a band of stromal coagulative necrosis and endothelial necrosis, with sparing of the anterior epithelium. In addition, there appeared some potential for damage to Descemet's membrane at the highest energy level tested. CONCLUSIONS: Findings indicate that the rabbit corneais subject to injury at the 1,318-nm wavelength with the established ED50.


Assuntos
Córnea/efeitos da radiação , Lasers/efeitos adversos , Lesões Experimentais por Radiação/patologia , Animais , Córnea/patologia , Opacidade da Córnea/etiologia , Opacidade da Córnea/patologia , Relação Dose-Resposta à Radiação , Feminino , Coelhos , Fatores de Tempo
7.
PLoS Negl Trop Dis ; 8(8): e2980, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25102067

RESUMO

BACKGROUND: Japanese encephalitis (JE) is a major cause of mortality and morbidity for which there is no treatment. In addition to direct viral cytopathology, the inflammatory response is postulated to contribute to the pathogenesis. Our goal was to determine the contribution of bystander effects and inflammatory mediators to neuronal cell death. METHODOLOGY/PRINCIPAL FINDINGS: Material from a macaque model was used to characterize the inflammatory response and cytopathic effects of JE virus (JEV). Intranasal JEV infection induced a non-suppurative encephalitis, dominated by perivascular, infiltrates of mostly T cells, alongside endothelial cell activation, vascular damage and blood brain barrier (BBB) leakage; in the adjacent parenchyma there was macrophage infiltration, astrocyte and microglia activation. JEV antigen was mostly in neurons, but there was no correlation between intensity of viral infection and degree of inflammatory response. Apoptotic cell death occurred in both infected and non-infected neurons. Interferon-α, which is a microglial activator, was also expressed by both. Tumour Necrosis Factor-α, inducible nitric oxide synthase and nitrotyrosine were expressed by microglial cells, astrocytes and macrophages. The same cells expressed matrix metalloproteinase (MMP)-2 whilst MMP-9 was expressed by neurons. CONCLUSIONS/SIGNIFICANCE: The results are consistent with JEV inducing neuronal apoptotic death and release of cytokines that initiate microglial activation and release of pro-inflammatory and apoptotic mediators with subsequent apoptotic death of both infected and uninfected neurons. Activation of astrocytes, microglial and endothelial cells likely contributes to inflammatory cell recruitment and BBB breakdown. It appears that neuronal apoptotic death and activation of microglial cells and astrocytes play a crucial role in the pathogenesis of JE.


Assuntos
Modelos Animais de Doenças , Encefalite Japonesa/etiologia , Animais , Apoptose , Citocinas/análise , Encefalite Japonesa/imunologia , Encefalite Japonesa/patologia , Macaca mulatta , Metaloproteinases da Matriz/análise , Neurônios/enzimologia , Neurônios/virologia , Óxido Nítrico/biossíntese
8.
Vaccine ; 29(35): 5925-31, 2011 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-21723353

RESUMO

Immune sera from volunteers vaccinated in a blinded Phase 3 clinical trial with JE-VAX(®) and a new Japanese encephalitis virus (JEV) vaccine (IC51 or IXIARO), were tested for the ability to protect mice against lethal JEV challenge. Sera from IXIARO vaccinated subjects were pooled into four batches based on neutralizing antibody measured by plaque reduction neutralization test (PRNT(50) titer): high (∼200), medium (∼40-50), low (∼20) and negative (<10). Pooled sera from JE-VAX(®) vaccinated subjects (PRNT(50) titer∼55) and pooled JEV antibody negative pre-vaccination sera were used as controls. Groups of ten 6- to 7-week-old female ICR mice were injected intraperitoneally with 0.5 ml of each serum pool diluted 1:2 or 1:10, challenged approximately 18 h later with a lethal dose of either JEV strain SA14 (genotype III) or strain KE-093 (genotype I) and observed for 21 days. All mice in the non-immune serum groups developed clinical signs consistent with JEV infection or died, whereas high titer sera from both IXIARO and JE-VAX(®) sera protected 90-100% of the animals. Statistical tests showed similar protection against both JEV strains SA14 and KE-093 and protection correlated with the anti-JEV antibody titer of IXIARO sera as measured by PRNT(50). Ex vivo neutralizing antibody titers showed that almost all mice with a titer of 10 or greater were fully protected. In a separate study, analysis of geometric mean titers (GMTs) of the groups of mice vaccinated with different doses of IXIARO and challenged with JEV SA14 provided additional evidence that titers≥10 were protective.


Assuntos
Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Vírus da Encefalite Japonesa (Espécie)/imunologia , Encefalite Japonesa/prevenção & controle , Vacinas contra Encefalite Japonesa/administração & dosagem , Animais , Vírus da Encefalite Japonesa (Espécie)/patogenicidade , Encefalite Japonesa/imunologia , Encefalite Japonesa/mortalidade , Encefalite Japonesa/virologia , Feminino , Humanos , Soros Imunes/administração & dosagem , Soros Imunes/imunologia , Imunização , Imunização Passiva , Vacinas contra Encefalite Japonesa/imunologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Testes de Neutralização , Análise de Sobrevida , Resultado do Tratamento
9.
Microb Pathog ; 33(5): 193-201, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12473434

RESUMO

Pneumocystis carinii pneumonia (PCP) is a frequent and serious opportunistic infection in immunocompromized patients. Although the pathogenesis of PCP-mediated lung injury is poorly understood, a central involvement of host inflammatory responses has been implicated. We have found that while the loss of specific T cell costimulatory signals increases susceptibility to the spontaneous pneumocystis infection, PCP-induced pulmonary injury (and subsequent morbidity and mortality) involves other intact costimulatory pathways. Mice that are genetically deficient for the costimulatory receptor CD154 (CD154 knockout (ko) mice) spontaneously developed PCP, consistent with the increased susceptibility of X-linked hyper IgM syndrome patients (caused by CD154 gene mutations) to P. carinii infection. In these mice PCP was manifested by progressive weight loss, dyspnea and death. In contrast, CD154 ko mice also genetically lacking ICAM1 (CD154 koxICAM1 ko) or CD28 (CD154 koxCD28 ko) costimulatory receptors had later onset of weight loss and significantly prolonged survival. Although onset of infection and age-matched P. carinii organism burden were equivalent, the CD154 single knockout mice had evidence of greater pulmonary inflammation vs. the double ko's. These findings suggest that costimulation-dependent T cell-mediated inflammation plays an important role in both susceptibility to and pathogenesis of PCP, and may identify potential molecular targets for novel immunomodulatory treatment approaches.


Assuntos
Pneumocystis/patogenicidade , Pneumonia por Pneumocystis/imunologia , Linfócitos T/imunologia , Animais , Peso Corporal , Antígenos CD28/imunologia , Ligante de CD40/genética , Ligante de CD40/imunologia , Imunidade Inata , Molécula 1 de Adesão Intercelular/imunologia , Leucócitos Mononucleares/imunologia , Ativação Linfocitária , Camundongos , Camundongos Knockout , Pneumonia por Pneumocystis/patologia , Baço/citologia , Baço/imunologia , Análise de Sobrevida
10.
Bol. Cent. Panamerican. Fiebre Aftosa ; (64/67): 18-25, 1998-2001. tab
Artigo em Inglês | LILACS | ID: lil-315233

RESUMO

Una tinción de inmunoperoxidasa indirecta (complejo avidina-biotina)usando el anticuerpo monoclonal 5DF12-3B6, que reconoce la proteína N del virus de la rabia, se usó para detectar antígeno del virus de la rabia en muestras de tejidos de 15 especies animales y una muestra humana infectadas con rabia naturalmente o experiementalmente. Este anticuerpo monoclonal reconoció todas las 16 cepas de virus de la rabia que se usaron en este estudio, como también lyssavirs relacionados a rabia como Duvenhage, Lagos Bat y Mokola. La muestra infectada com Mokola inicialmente sólo demonstró una tinción débil, la que se hizo más fuerte cundo se eliminó el tratamiento con Pronase E. La tinción es sensible y específica, identificando correctamente al antígeno de rabia en todas las muestras usadas con excepción de una muestra (37/38) que era débilmente positiva por IFA y muy pequeña. Además no se detectó tinción específica en las muestras negativas (23/23). La utilidad del método de tinción de inmunohistoquímica descrito se base en la habilidad de un anticuerpo monoclonal para reconocer un amplio espectro de lyssavirus en tejidos fijados en formalina


Assuntos
Anticorpos Monoclonais , Imuno-Histoquímica , Lyssavirus , Raiva , Vírus da Raiva
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