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  1 / 4164 MEDLINE  
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[PMID]:29420464
[Au] Autor:Vora NM; Orciari LA; Bertumen JB; Damon I; Ellison JA; Fowler VG; Franka R; Petersen BW; Satheshkumar PS; Schexnayder SM; Smith TG; Wallace RM; Weinstein S; Williams C; Yager P; Niezgoda M
[Ti] Título:Potential Confounding of Diagnosis of Rabies in Patients with Recent Receipt of Intravenous Immune Globulin.
[So] Source:MMWR Morb Mortal Wkly Rep;67(5):161-165, 2018 Feb 09.
[Is] ISSN:1545-861X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Rabies is an acute encephalitis that is nearly always fatal. It is caused by infection with viruses of the genus Lyssavirus, the most common of which is Rabies lyssavirus. The Council of State and Territorial Epidemiologists (CSTE) defines a confirmed human rabies case as an illness compatible with rabies that meets at least one of five different laboratory criteria.* Four of these criteria do not depend on the patient's rabies vaccination status; however, the remaining criterion, "identification of Lyssavirus-specific antibody (i.e. by indirect fluorescent antibody…test or complete [Rabies lyssavirus] neutralization at 1:5 dilution) in the serum," is only considered diagnostic in unvaccinated patients. Lyssavirus-specific antibodies include Rabies lyssavirus-specific binding immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies and Rabies lyssavirus neutralizing antibodies (RLNAs). This report describes six patients who were tested for rabies by CDC and who met CSTE criteria for confirmed human rabies because they had illnesses compatible with rabies, had not been vaccinated for rabies, and were found to have serum RLNAs (with complete Rabies lyssavirus neutralization at a serum dilution of 1:5). An additional four patients are described who were tested for rabies by CDC who were found to have serum RLNAs (with incomplete Rabies lyssavirus neutralization at a serum dilution of 1:5) despite having not been vaccinated for rabies. None of these 10 patients received a rabies diagnosis; rather, they were considered to have been passively immunized against rabies through recent receipt of intravenous immune globulin (IVIG). Serum RLNA test results should be interpreted with caution in patients who have not been vaccinated against rabies but who have recently received IVIG.
[Mh] Termos MeSH primário: Imunoglobulinas Intravenosas/administração & dosagem
Raiva/diagnóstico
[Mh] Termos MeSH secundário: Adolescente
Adulto
Criança
Reações Falso-Positivas
Feminino
Seres Humanos
Imunização Passiva
Lyssavirus/isolamento & purificação
Masculino
Meia-Idade
Vacinas Antirrábicas/administração & dosagem
Vírus da Raiva/isolamento & purificação
Adulto Jovem
[Pt] Tipo de publicação:CASE REPORTS; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Immunoglobulins, Intravenous); 0 (Rabies Vaccines)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180222
[Lr] Data última revisão:
180222
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180209
[St] Status:MEDLINE
[do] DOI:10.15585/mmwr.mm6705a3


  2 / 4164 MEDLINE  
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[PMID]:29261658
[Au] Autor:Chao TY; Ren S; Shen E; Moore S; Zhang SF; Chen L; Rupprecht CE; Tsao E
[Ad] Endereço:Synermore Biologics Co., Ltd., Taipei, Taiwan.
[Ti] Título:SYN023, a novel humanized monoclonal antibody cocktail, for post-exposure prophylaxis of rabies.
[So] Source:PLoS Negl Trop Dis;11(12):e0006133, 2017 12.
[Is] ISSN:1935-2735
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Rabies is a neglected zoonotic disease that is preventable in humans by appropriate post-exposure prophylaxis (PEP). However, current PEP relies on polyclonal immune globulin products purified from pooled human (HRIG) or equine (ERIG) plasma that are either in chronic shortage or in association with safety concerns. Here, we present the development of an antibody cocktail, SYN023, made of two novel monoclonal antibodies (MAb) CTB011 and CTB012 that could serve as safer and more cost-effective alternatives to the current RIG products. Both CTB011 and CTB012 are humanized MAbs that bind to non-overlapping epitopes on the rabies virus (RABV) glycoprotein (G) with sub-nanomolar affinities. Sequence analysis revealed that many of the critical residues in binding are highly conserved across different species of lyssaviruses. When combined at a 1:1 ratio, CTB011/CTB012 exhibited neutralization capabilities equivalent or superior to HRIG against 10 North American street RABV isolates in vitro and 15 prevalent Chinese RABV strains in animal models. Finally, SYN023, at a dosage of 0.03 mg/kg, was able to offer the same degree of protection as standard HRIG administration (20 IU/kg) in Syrian hamsters challenged with a highly virulent bat (Tadarida brasiliensis) RABV variant. Taken together, the high-potency and broad-spectrum neutralization demonstrated by SYN023 make it an effective candidate for human rabies PEP consideration.
[Mh] Termos MeSH primário: Anticorpos Monoclonais/imunologia
Anticorpos Antivirais/imunologia
Profilaxia Pós-Exposição
Vírus da Raiva/imunologia
Raiva/prevenção & controle
[Mh] Termos MeSH secundário: Animais
Anticorpos Monoclonais Humanizados/imunologia
Quirópteros
Feminino
Glicoproteínas/imunologia
Seres Humanos
Mesocricetus
Camundongos
Camundongos Endogâmicos BALB C
Raiva/virologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antibodies, Monoclonal); 0 (Antibodies, Monoclonal, Humanized); 0 (Antibodies, Viral); 0 (Glycoproteins)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180212
[Lr] Data última revisão:
180212
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171221
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pntd.0006133


  3 / 4164 MEDLINE  
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[PMID]:29267277
[Au] Autor:Um J; Chun BC; Lee YS; Hwang KJ; Yang DK; Park JS; Kim SY
[Ad] Endereço:Division of Zoonoses, Center for Immunology & Pathology, Korea National Institute of Health, Cheongju-si, Chungcheongbuk-do, Republic of Korea.
[Ti] Título:Development and evaluation of an anti-rabies virus phosphoprotein-specific monoclonal antibody for detection of rabies neutralizing antibodies using RFFIT.
[So] Source:PLoS Negl Trop Dis;11(12):e0006084, 2017 12.
[Is] ISSN:1935-2735
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Rabies is a major public health problem with a fatality rate close to 100%; however, complete prevention can be achieved through pre- or post-exposure prophylaxis. The rapid fluorescent focus inhibition test (RFFIT) is one of the recommended testing methods to determine the production of neutralizing antibodies after vaccination. Here, we report the development of a new monoclonal antibody (mAb) designed to react specifically with Rabies virus (RABV) phosphoprotein (P protein), and the evaluation of its applicability to the RFFIT and its effectiveness as a diagnostic reagent for human rabies. METHODOLOGY/PRINCIPAL FINDINGS: The mAb KGH P 16B8 was produced to target the P protein of the Korean KGH RABV strain. An indirect immunofluorescence assay (IFA) was conducted to detect various strains of RABV in various cell lines. Alexa-conjugated KGH P 16B8 (16B8-Alexa) was developed for the RFFIT. The IFA test could detect RABV up to a 1:2,500 dilution, with a detection limit comparable to that of a commercial diagnostic reagent. The sensitivity, specificity, positive predictive value, and negative predictive value of the RFFIT using 16B8-Alexa in 414 clinical specimens were 98.67%, 99.47%, 99.55%, and 98.42%, respectively. The results of the RFFIT with 16B8-Alexa were strongly correlated with those obtained using an existing commercial diagnostic reagent (r = 0.995, p<0.001). CONCLUSIONS/SIGNIFICANCE: The mAb developed in this study shows high sensitivity and specificity, confirming its clinical utility with the RFFIT to measure the rabies neutralizing antibody titer and establish a diagnosis in human. Thus, 16B8-Alexa is expected to serve as an alternative diagnostic reagent that is widely accessible, with potentially broad applications beyond those of the RFFIT in Korea. Further studies with 16B8-Alexa should provide insight into the immunological mechanism of the P protein of Korean RABV.
[Mh] Termos MeSH primário: Anticorpos Monoclonais/imunologia
Anticorpos Neutralizantes/sangue
Anticorpos Antivirais/sangue
Técnica Indireta de Fluorescência para Anticorpo/métodos
Testes de Neutralização/métodos
Fosfoproteínas/imunologia
Vírus da Raiva/imunologia
Raiva/prevenção & controle
Proteínas Estruturais Virais/imunologia
[Mh] Termos MeSH secundário: Anticorpos Neutralizantes/imunologia
Anticorpos Antivirais/imunologia
Profilaxia Pós-Exposição/métodos
Raiva/virologia
Vacinas Antirrábicas/imunologia
Vacinação
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antibodies, Monoclonal); 0 (Antibodies, Neutralizing); 0 (Antibodies, Viral); 0 (P phosphoprotein, Rabies virus); 0 (Phosphoproteins); 0 (Rabies Vaccines); 0 (Viral Structural Proteins)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180210
[Lr] Data última revisão:
180210
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171222
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pntd.0006084


  4 / 4164 MEDLINE  
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[PMID]:29323857
[Au] Autor:Dedkov VG; Deviatkin AA; Poleschuk EM; Safonova MV; Markelov ML; Shipulin GA
[Ti] Título:Development and evaluation of the RT-PCR kit for the rabies virus diagnosis.
[So] Source:Vopr Virusol;61(5):235-40, 2016.
[Is] ISSN:0507-4088
[Cp] País de publicação:Russia (Federation)
[La] Idioma:eng
[Ab] Resumo:To improve the diagnosis, surveillance, and control for the rabies virus, a kit for hybridization-triggered fluorescence detection of rabies virus DNA by the RT-PCR technique was developed and evaluated. The analytical sensitivity of the kit was 4*10 GE per ml. High specificity of the kit was shown using representative sampling of viral, bacterial, and human nucleic acids.
[Mh] Termos MeSH primário: RNA Viral/genética
Vírus da Raiva/genética
Raiva/epidemiologia
Raiva/veterinária
Reação em Cadeia da Polimerase em Tempo Real/normas
Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas
[Mh] Termos MeSH secundário: Animais
Gatos
Primers do DNA/síntese química
Primers do DNA/genética
Cervos/virologia
Cães
Raposas/virologia
Seres Humanos
Raiva/diagnóstico
Raiva/transmissão
Vírus da Raiva/isolamento & purificação
Reação em Cadeia da Polimerase em Tempo Real/métodos
Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos
Federação Russa/epidemiologia
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Primers); 0 (RNA, Viral)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180208
[Lr] Data última revisão:
180208
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180112
[St] Status:MEDLINE


  5 / 4164 MEDLINE  
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[PMID]:28461570
[Au] Autor:Bongiorno EK; Garcia SA; Sauma S; Hooper DC
[Ad] Endereço:Department of Cancer Biology, Thomas Jefferson University, Philadelphia, PA 19107; and.
[Ti] Título:Type 1 Immune Mechanisms Driven by the Response to Infection with Attenuated Rabies Virus Result in Changes in the Immune Bias of the Tumor Microenvironment and Necrosis of Mouse GL261 Brain Tumors.
[So] Source:J Immunol;198(11):4513-4523, 2017 06 01.
[Is] ISSN:1550-6606
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Immunotherapeutic strategies for malignant glioma have to overcome the immunomodulatory activities of M2 monocytes that appear in the circulation and as tumor-associated macrophages (TAMs). M2 cell products contribute to the growth-promoting attributes of the tumor microenvironment (TME) and bias immunity toward type 2, away from the type 1 mechanisms with antitumor properties. To drive type 1 immunity in CNS tissues, we infected GL261 tumor-bearing mice with attenuated rabies virus (RABV). These neurotropic viruses spread to CNS tissues -axonally, where they induce a strong type 1 immune response that involves Th1, CD8, and B cell entry across the blood-brain barrier and virus clearance in the absence of overt sequelae. Intranasal infection with attenuated RABV prolonged the survival of mice bearing established GL261 brain tumors. Despite the failure of virus spread to the tumor, infection resulted in significantly enhanced tumor necrosis, extensive CD4 T cell accumulation, and high levels of the proinflammatory factors IFN-γ, TNF-α, and inducible NO synthase in the TME merely 4 d postinfection, before significant virus spread or the appearance of RABV-specific immune mechanisms in CNS tissues. Although the majority of infiltrating CD4 cells appeared functionally inactive, the proinflammatory changes in the TME later resulted in the loss of accumulating M2 and increased M1 TAMs. Mice deficient in the Th1 transcription factor T-bet did not gain any survival advantage from RABV infection, exhibiting only limited tumor necrosis and no change in TME cytokines or TAM phenotype and highlighting the importance of type 1 mechanisms in this process.
[Mh] Termos MeSH primário: Neoplasias Encefálicas/imunologia
Neoplasias Encefálicas/patologia
Encéfalo/virologia
Vírus da Raiva/imunologia
Microambiente Tumoral/imunologia
[Mh] Termos MeSH secundário: Animais
Linfócitos B/imunologia
Barreira Hematoencefálica/imunologia
Barreira Hematoencefálica/patologia
Barreira Hematoencefálica/virologia
Encéfalo/imunologia
Neoplasias Encefálicas/virologia
Linfócitos T CD4-Positivos
Citocinas/imunologia
Modelos Animais de Doenças
Feminino
Interferon gama/biossíntese
Interferon gama/imunologia
Camundongos
Necrose/virologia
Óxido Nítrico Sintase Tipo II/biossíntese
Óxido Nítrico Sintase Tipo II/metabolismo
Vírus da Raiva/genética
Vírus da Raiva/fisiologia
Proteínas com Domínio T-Box/deficiência
Proteínas com Domínio T-Box/metabolismo
Células Th2/imunologia
Fator de Necrose Tumoral alfa/biossíntese
Fator de Necrose Tumoral alfa/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (Cytokines); 0 (T-Box Domain Proteins); 0 (T-box transcription factor TBX21); 0 (Tumor Necrosis Factor-alpha); 82115-62-6 (Interferon-gamma); EC 1.14.13.39 (Nitric Oxide Synthase Type II)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:180127
[Lr] Data última revisão:
180127
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE
[do] DOI:10.4049/jimmunol.1601444


  6 / 4164 MEDLINE  
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[PMID]:29216187
[Au] Autor:Stitz L; Vogel A; Schnee M; Voss D; Rauch S; Mutzke T; Ketterer T; Kramps T; Petsch B
[Ad] Endereço:Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
[Ti] Título:A thermostable messenger RNA based vaccine against rabies.
[So] Source:PLoS Negl Trop Dis;11(12):e0006108, 2017 Dec.
[Is] ISSN:1935-2735
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Although effective rabies virus vaccines have been existing for decades, each year, rabies virus infections still cause around 50.000 fatalities worldwide. Most of these cases occur in developing countries, where these vaccines are not available. The reasons for this are the prohibitive high costs of cell culture or egg grown rabies virus vaccines and the lack of a functional cold chain in many regions in which rabies virus is endemic. Here, we describe the excellent temperature resistance of a non-replicating mRNA based rabies virus vaccine encoding the rabies virus glycoprotein (RABV-G). Prolonged storage of the vaccine from -80°C to up to +70°C for several months did not impact the protective capacity of the mRNA vaccine. Efficacy after storage was demonstrated by the induction of rabies specific virus neutralizing antibodies and protection in mice against lethal rabies infection. Moreover, storing the vaccine at oscillating temperatures between +4° and +56°C for 20 cycles in order to simulate interruptions of the cold chain during vaccine transport, did not affect the vaccine's immunogenicity and protective characteristics, indicating that maintenance of a cold chain is not essential for this vaccine.
[Mh] Termos MeSH primário: Antígenos Virais/genética
Glicoproteínas/genética
Imunogenicidade da Vacina
RNA Mensageiro
Vacinas Antirrábicas/imunologia
Vírus da Raiva/genética
Raiva/prevenção & controle
Potência de Vacina
Proteínas do Envelope Viral/genética
[Mh] Termos MeSH secundário: Animais
Anticorpos Neutralizantes/sangue
Anticorpos Antivirais/sangue
Camundongos
Vacinas Antirrábicas/administração & dosagem
Vacinas Antirrábicas/genética
Vírus da Raiva/imunologia
Temperatura Ambiente
Vacinas Sintéticas/administração & dosagem
Vacinas Sintéticas/genética
Vacinas Sintéticas/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Neutralizing); 0 (Antibodies, Viral); 0 (Antigens, Viral); 0 (Glycoproteins); 0 (RNA, Messenger); 0 (Rabies Vaccines); 0 (Vaccines, Synthetic); 0 (Viral Envelope Proteins); 0 (glycoprotein G, Rabies virus)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171226
[Lr] Data última revisão:
171226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171208
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pntd.0006108


  7 / 4164 MEDLINE  
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[PMID]:27773481
[Au] Autor:Xu C; Krabbe S; Gründemann J; Botta P; Fadok JP; Osakada F; Saur D; Grewe BF; Schnitzer MJ; Callaway EM; Lüthi A
[Ad] Endereço:Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, 4058 Basel, Switzerland.
[Ti] Título:Distinct Hippocampal Pathways Mediate Dissociable Roles of Context in Memory Retrieval.
[So] Source:Cell;167(4):961-972.e16, 2016 11 03.
[Is] ISSN:1097-4172
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Memories about sensory experiences are tightly linked to the context in which they were formed. Memory contextualization is fundamental for the selection of appropriate behavioral reactions needed for survival, yet the underlying neuronal circuits are poorly understood. By combining trans-synaptic viral tracing and optogenetic manipulation, we found that the ventral hippocampus (vHC) and the amygdala, two key brain structures encoding context and emotional experiences, interact via multiple parallel pathways. A projection from the vHC to the basal amygdala mediates fear behavior elicited by a conditioned context, whereas a parallel projection from a distinct subset of vHC neurons onto midbrain-projecting neurons in the central amygdala is necessary for context-dependent retrieval of cued fear memories. Our findings demonstrate that two fundamentally distinct roles of context in fear memory retrieval are processed by distinct vHC output pathways, thereby allowing for the formation of robust contextual fear memories while preserving context-dependent behavioral flexibility.
[Mh] Termos MeSH primário: Tonsila do Cerebelo/fisiologia
Hipocampo/fisiologia
Memória
Vias Neurais
[Mh] Termos MeSH secundário: Animais
Condicionamento (Psicologia)
Fenômenos Eletrofisiológicos
Medo
Camundongos
Camundongos Endogâmicos C57BL
Neurônios/citologia
Neurônios/fisiologia
Optogenética
Vírus da Raiva/genética
Sinapses
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1704
[Cu] Atualização por classe:171127
[Lr] Data última revisão:
171127
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161105
[St] Status:MEDLINE


  8 / 4164 MEDLINE  
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[PMID]:29045436
[Au] Autor:Kim PK; Keum SJ; Osinubi MOV; Franka R; Shin JY; Park ST; Kim MS; Park MJ; Lee SY; Carson W; Greenberg L; Yu P; Tao X; Lihua W; Tang Q; Liang G; Shampur M; Rupprecht CE; Chang SJ
[Ad] Endereço:Celltrion, INC, Department of Research and Development, Incheon, Republic of Korea.
[Ti] Título:Development and characterization of novel chimeric monoclonal antibodies for broad spectrum neutralization of rabies virus.
[So] Source:PLoS One;12(10):e0186380, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Current post-exposure prophylaxis for rabies virus infection has several limitations in terms of supply, cost, safety, and efficacy. Attempts to replace human or equine rabies immune globulins (HRIG or ERIG) have been made by several companies and institutes. We developed potent monoclonal antibodies to neutralize a broad spectrum of rabies viruses by screening hybridomas received from the U.S. Centers for Disease Control and Prevention (CDC). Two kinds of chimeric human antibodies (chimeric #7 and #17) were constructed by cloning the variable regions from selected hybridomas and the constant region of a human antibody. Two antibodies were bound to antigenic site III and I/IV, respectively, and were able to neutralize 51 field isolates of rabies virus that were isolated at different times and places such as Asia, Africa, North America, South America, and Australia. These two antibodies neutralize rabies viruses with high efficacy in an in vivo test using Syrian hamster and mouse models and show low risk for adverse immunogenicity.
[Mh] Termos MeSH primário: Anticorpos Monoclonais/administração & dosagem
Anticorpos Neutralizantes/administração & dosagem
Vírus da Raiva/imunologia
Raiva/prevenção & controle
[Mh] Termos MeSH secundário: África
Animais
Anticorpos Monoclonais/genética
Anticorpos Monoclonais/imunologia
Anticorpos Neutralizantes/genética
Anticorpos Neutralizantes/imunologia
Ásia
Austrália
Centers for Disease Control and Prevention (U.S.)
Modelos Animais de Doenças
Cavalos/virologia
Seres Humanos
Hibridomas/imunologia
Mesocricetus/virologia
Camundongos
América do Norte
Raiva/imunologia
Raiva/virologia
Vírus da Raiva/patogenicidade
América do Sul
Estados Unidos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Monoclonal); 0 (Antibodies, Neutralizing)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171031
[Lr] Data última revisão:
171031
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171019
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0186380


  9 / 4164 MEDLINE  
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[PMID]:28981124
[Au] Autor:Newton R; Delguste M; Koehler M; Dumitru AC; Laskowski PR; Müller DJ; Alsteens D
[Ad] Endereço:Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.
[Ti] Título:Combining confocal and atomic force microscopy to quantify single-virus binding to mammalian cell surfaces.
[So] Source:Nat Protoc;12(11):2275-2292, 2017 Nov.
[Is] ISSN:1750-2799
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Over the past five years, atomic force microscopy (AFM)-based approaches have evolved into a powerful multiparametric tool set capable of imaging the surfaces of biological samples ranging from single receptors to membranes and tissues. One of these approaches, force-distance curve-based AFM (FD-based AFM), uses a probing tip functionalized with a ligand to image living cells at high-resolution and simultaneously localize and characterize specific ligand-receptor binding events. Analyzing data from FD-based AFM experiments using appropriate probabilistic models allows quantification of the kinetic and thermodynamic parameters that describe the free-energy landscape of the ligand-receptor bond. We have recently developed an FD-based AFM approach to quantify the binding events of single enveloped viruses to surface receptors of living animal cells while simultaneously observing them by fluorescence microscopy. This approach has provided insights into the early stages of the interaction between a virus and a cell. Applied to a model virus, we probed the specific interaction with cells expressing viral cognate receptors and measured the affinity of the interaction. Furthermore, we observed that the virus rapidly established specific multivalent interactions and found that each bond formed in sequence strengthened the attachment of the virus to the cell. Here we describe detailed procedures for probing the specific interactions of viruses with living cells; these procedures cover tip preparation, cell sample preparation, step-by-step FD-based AFM imaging and data analysis. Experienced microscopists should be able to master the entire set of protocols in 1 month.
[Mh] Termos MeSH primário: Membrana Celular/metabolismo
Microscopia de Força Atômica/métodos
Microscopia Confocal/métodos
Vírus da Raiva/metabolismo
Ligação Viral
[Mh] Termos MeSH secundário: Animais
Membrana Celular/ultraestrutura
Cães
Células HEK293
Seres Humanos
Células Madin Darby de Rim Canino
Microscopia de Força Atômica/instrumentação
Microscopia Confocal/instrumentação
Vírus da Raiva/ultraestrutura
Vesiculovirus
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171018
[Lr] Data última revisão:
171018
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171006
[St] Status:MEDLINE
[do] DOI:10.1038/nprot.2017.112


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[PMID]:28951448
[Au] Autor:Albisetti GW; Ghanem A; Foster E; Conzelmann KK; Zeilhofer HU; Wildner H
[Ad] Endereço:Institute of Pharmacology and Toxicology, University of Zurich, CH-8057 Zürich, Switzerland.
[Ti] Título:Identification of Two Classes of Somatosensory Neurons That Display Resistance to Retrograde Infection by Rabies Virus.
[So] Source:J Neurosci;37(43):10358-10371, 2017 Oct 25.
[Is] ISSN:1529-2401
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Glycoprotein-deleted rabies virus-mediated monosynaptic tracing has become a standard method for neuronal circuit mapping, and is applied to virtually all parts of the rodent nervous system, including the spinal cord and primary sensory neurons. Here we identified two classes of unmyelinated sensory neurons (nonpeptidergic and C-fiber low-threshold mechanoreceptor neurons) resistant to direct and trans-synaptic infection from the spinal cord with rabies viruses that carry glycoproteins in their envelopes and that are routinely used for infection of CNS neurons (SAD-G and N2C-G). However, the same neurons were susceptible to infection with EnvA-pseudotyped rabies virus in tumor virus A receptor transgenic mice, indicating that resistance to retrograde infection was due to impaired virus adsorption rather than to deficits in subsequent steps of infection. These results demonstrate an important limitation of rabies virus-based retrograde tracing of sensory neurons in adult mice, and may help to better understand the molecular machinery required for rabies virus spread in the nervous system. In this study, mice of both sexes were used. To understand the neuronal bases of behavior, it is important to identify the underlying neural circuitry. Rabies virus-based monosynaptic tracing has been used to identify neuronal circuits in various parts of the nervous system. This has included connections between peripheral sensory neurons and their spinal targets. These connections form the first synapse in the somatosensory pathway. Here we demonstrate that two classes of unmyelinated sensory neurons, which account for >40% of dorsal root ganglia neurons, display resistance to rabies infection. Our results are therefore critical for interpreting monosynaptic rabies-based tracing in the sensory system. In addition, identification of rabies-resistant neurons might provide a means for future studies addressing rabies pathobiology.
[Mh] Termos MeSH primário: Gânglios Espinais/química
Rede Nervosa/química
Técnicas de Rastreamento Neuroanatômico/métodos
Vírus da Raiva
Células Receptoras Sensoriais/química
[Mh] Termos MeSH secundário: Animais
Feminino
Gânglios Espinais/citologia
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Camundongos Transgênicos
Rede Nervosa/citologia
Células do Corno Posterior/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171115
[Lr] Data última revisão:
171115
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170928
[St] Status:MEDLINE
[do] DOI:10.1523/JNEUROSCI.1277-17.2017



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