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[PMID]:28986291
[Au] Autor:Meli ML; Berger A; Willi B; Spiri AM; Riond B; Hofmann-Lehmann R
[Ad] Endereço:Clinical Laboratory, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland; Center for Clinical Studies, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland. Electronic address: mmeli@vetclinics.uzh.ch.
[Ti] Título:Molecular detection of feline calicivirus in clinical samples: A study comparing its detection by RT-qPCR directly from swabs and after virus isolation.
[So] Source:J Virol Methods;251:54-60, 2018 Jan.
[Is] ISSN:1879-0984
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Feline caliciviruses (FCVs) are non-enveloped RNA viruses that exhibit high genetic variation. Two reverse transcription quantitative polymerase chain reaction (RT-qPCR) FCV assays (S1 and S2) were evaluated using samples from 300 field cats. The direct detection of FCV in swabs and after propagation in cell culture, as well as the influence of storage conditions, was assessed. FCV-RNA detectability on dry swabs was similar after storage at either 4°C or -20°C, but viral burdens were maintained for a longer time period when viral transport medium was used. A total of 97 (32%) samples was considered FCV PCR-positive. Of these, 81% and 77% tested positive directly from swabs using S1 and S2, respectively; 84% and 81% tested positive after enrichment in cell culture, respectively. Combined detection by RT-PCR directly from swabs and after VI was most sensitive (up to 96%). Neither of the methods alone were able to detect all FCV-positive samples. In conclusion, clinical samples should be collected in viral transport medium, stored at ≤4°C and processed as soon as possible. The combination of cell culture with RT-qPCR or detection directly from swabs using a combination of different RT-qPCR assays is recommended to reach a high sensitivity of FCV detection.
[Mh] Termos MeSH primário: Infecções por Caliciviridae/veterinária
Calicivirus Felino/isolamento & purificação
Doenças do Gato/diagnóstico
Doenças do Gato/virologia
Técnicas de Diagnóstico Molecular/métodos
Reação em Cadeia da Polimerase em Tempo Real/métodos
Manejo de Espécimes/métodos
[Mh] Termos MeSH secundário: Animais
Infecções por Caliciviridae/diagnóstico
Infecções por Caliciviridae/virologia
Gatos
Sensibilidade e Especificidade
Cultura de Vírus
[Pt] Tipo de publicação:COMPARATIVE STUDY; EVALUATION STUDIES; JOURNAL ARTICLE
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180201
[Lr] Data última revisão:
180201
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171008
[St] Status:MEDLINE


  2 / 10500 MEDLINE  
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[PMID]:28468878
[Au] Autor:Gravel A; Dubuc I; Wallaschek N; Gilbert-Girard S; Collin V; Hall-Sedlak R; Jerome KR; Mori Y; Carbonneau J; Boivin G; Kaufer BB; Flamand L
[Ad] Endereço:Division of Infectious and Immune Diseases, CHU de Quebec Research Center, Quebec City, Canada.
[Ti] Título:Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration.
[So] Source:J Virol;91(14), 2017 Jul 15.
[Is] ISSN:1098-5514
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Human herpesviruses 6A/B (HHV-6A/B) can integrate their viral genomes in the telomeres of human chromosomes. The viral and cellular factors contributing to HHV-6A/B integration remain largely unknown, mostly due to the lack of efficient and reproducible cell culture models to study HHV-6A/B integration. In this study, we characterized the HHV-6A/B integration efficiencies in several human cell lines using two different approaches. First, after a short-term infection (5 h), cells were processed for single-cell cloning and analyzed for chromosomally integrated HHV-6A/B (ciHHV-6A/B). Second, cells were infected with HHV-6A/B and allowed to grow in bulk for 4 weeks or longer and then analyzed for the presence of ciHHV-6. Using quantitative PCR (qPCR), droplet digital PCR, and fluorescent hybridization, we could demonstrate that HHV-6A/B integrated in most human cell lines tested, including telomerase-positive (HeLa, MCF-7, HCT-116, and HEK293T) and telomerase-negative cell lines (U2OS and GM847). Our results also indicate that inhibition of DNA replication, using phosphonoacetic acid, did not affect HHV-6A/B integration. Certain clones harboring ciHHV-6A/B spontaneously express viral genes and proteins. Treatment of cells with phorbol ester or histone deacetylase inhibitors triggered the expression of many viral genes, including , , and , without the production of infectious virus, suggesting that the tested stimuli were not sufficient to trigger full reactivation. In summary, both integration models yielded comparable results and should enable the identification of viral and cellular factors contributing to HHV-6A/B integration and the screening of drugs influencing viral gene expression, as well as the release of infectious HHV-6A/B from the integrated state. The analysis and understanding of HHV-6A/B genome integration into host DNA is currently limited due to the lack of reproducible and efficient viral integration systems. In the present study, we describe two quantitative cell culture viral integration systems. These systems can be used to define cellular and viral factors that play a role in HHV-6A/B integration. Furthermore, these systems will allow us to decipher the conditions resulting in virus gene expression and excision of the integrated viral genome resulting in reactivation.
[Mh] Termos MeSH primário: Herpesvirus Humano 6/fisiologia
Cultura de Vírus/métodos
Integração Viral
[Mh] Termos MeSH secundário: Técnicas de Cultura de Células/métodos
Linhagem Celular
Seres Humanos
Hibridização in Situ Fluorescente
Reação em Cadeia da Polimerase em Tempo Real
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171226
[Lr] Data última revisão:
171226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE


  3 / 10500 MEDLINE  
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[PMID]:27775612
[Au] Autor:von Nordheim M; Boinay M; Leisi R; Kempf C; Ros C
[Ad] Endereço:Department of Chemistry and Biochemistry, University of Bern, Bern 3012, Switzerland. marcus.vonnordheim@dcb.unibe.ch.
[Ti] Título:Cutthroat Trout Virus-Towards a Virus Model to Support Hepatitis E Research.
[So] Source:Viruses;8(10), 2016 10 20.
[Is] ISSN:1999-4915
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:Cutthroat trout virus (CTV) is a non-pathogenic fish virus belonging to the family, and it is distantly related to hepatitis E virus (HEV). Here, we report the development of an efficient cell culture system where CTV can consistently replicate to titers never observed before with a hepevirus. By using the rainbow trout gill (RTGill-W1) cell line, CTV reaches 10 geq/mL intracellularly and 108 geq/mL extracellularly within 5-6 days in culture. We additionally established a qPCR system to investigate CTV infectivity, and developed a specific antibody directed against the viral capsid protein encoded by ORF2. With these methods, we were able to follow the progressive accumulation of viral RNA and the capsid protein, and their intracellular distribution during virus replication. Virus progeny purified through iodixanol density gradients indicated-that similar to HEV-CTV produced in cell culture is also lipid-associated. The lack of an efficient cell culture system has greatly impeded studies with HEV, a major human pathogen that causes hepatitis worldwide. Although several cell culture systems have recently been established, the replication efficiency of HEV is not robust enough to allow studies on different aspects of the virus replication cycle. Therefore, a surrogate virus that can replicate easily and efficiently in cultured cells would be helpful to boost research studies with hepeviruses. Due to its similarities, but also its key differences to HEV, CTV represents a promising tool to elucidate aspects of the replication cycle of in general, and HEV in particular.
[Mh] Termos MeSH primário: Hepevirus/fisiologia
Cultura de Vírus/métodos
Replicação Viral
[Mh] Termos MeSH secundário: Animais
Antígenos Virais/análise
Linhagem Celular
Imunoensaio/métodos
Oncorhynchus mykiss
RNA Viral/análise
Reação em Cadeia da Polimerase em Tempo Real
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antigens, Viral); 0 (RNA, Viral)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:171212
[Lr] Data última revisão:
171212
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161025
[St] Status:MEDLINE


  4 / 10500 MEDLINE  
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[PMID]:28884263
[Au] Autor:Prezioso C; Scribano D; Bellizzi A; Anzivino E; Rodio DM; Trancassini M; Palamara AT; Pietropaolo V
[Ad] Endereço:Department of Public Health and Infectious Diseases, "Sapienza" University, P.le Aldo Moro, 5, 00185, Rome, Italy.
[Ti] Título:Efficient propagation of archetype JC polyomavirus in COS-7 cells: evaluation of rearrangements within the NCCR structural organization after transfection.
[So] Source:Arch Virol;162(12):3745-3752, 2017 Dec.
[Is] ISSN:1432-8798
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:John Cunningham virus (JCPyV) is an ubiquitous human pathogen that causes disease in immunocompromised patients. The JCPyV genome is composed of an early region and a late region, which are physically separated by the non-coding control region (NCCR). The DNA sequence of the NCCR distinguishes two forms of JCPyV, the designated archetype and the prototype, which resulted from a rearrangement of the archetype sequence. To date, the cell culture systems for propagating JCPyV archetype have been very limited in their availability and robustness. Prior to this study, it was demonstrated that JCPyV archetype DNA replicates in COS-7 simian kidney cells expressing SV40 TAg and COS-7 cells expressing HIV-1 Tat. Based on these observations, the present study was conducted to reproduce an in vitro model in COS-7 cells transfected with the JCPyV archetype strain in order to study JCPyV DNA replication and analyze NCCR rearrangements during the viral life cycle. The efficiency of JCPyV replication was evaluated by quantitative PCR (Q-PCR) and by hemagglutination (HA) assay after transfection. In parallel, sequence analysis of JCPyV NCCR was performed. JCPyV efficiently replicated in kidney-derived COS-7 cells, as demonstrated by a progressive increase in viral load and virion particle production after transfection. The archetypal structure of NCCR was maintained during the viral cycle, but two characteristic point mutations were detected 28 days after transfection. This model is a useful tool for analyzing NCCR rearrangements during in vitro replication in cells that are sites of viral persistence, such as tubular epithelial cells of the kidney.
[Mh] Termos MeSH primário: Adaptação Biológica
Rearranjo Gênico
Vírus JC/crescimento & desenvolvimento
Vírus JC/genética
[Mh] Termos MeSH secundário: Animais
Células COS
Cercopithecus aethiops
Testes de Hemaglutinação
Seres Humanos
Mutação Puntual
Reação em Cadeia da Polimerase em Tempo Real
Análise de Sequência de DNA
Transfecção
Cultura de Vírus
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171119
[Lr] Data última revisão:
171119
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170909
[St] Status:MEDLINE
[do] DOI:10.1007/s00705-017-3542-7


  5 / 10500 MEDLINE  
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[PMID]:28865204
[Au] Autor:Panneum S; Rukkwamsuk T
[Ad] Endereço:.
[Ti] Título:Diagnosis of Caprine Arthritis Encephalitis Virus infection in dairy goats by ELISA, PCR and Viral Culture.
[So] Source:Pol J Vet Sci;20(2):347-353, 2017 Mar 01.
[Is] ISSN:1505-1773
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:For preventive and control strategies of Caprine Arthritis Encephalitis Virus (CAEV) infection in dairy goats, performance of the available diagnostic tests was described as one of the most important and necessary aspects. The study aimed at evaluating the diagnostic test performance, including PCR, ELISA and viral culture, for CAEV infection in dairy goats in Thailand. Blood samples of 29 dairy goats from five low- to medium-prevalence herds and one very low-prevalence herd were collected for PCR and ELISA methods. The performance of these two diagnostic methods was evaluated by comparing with cytopathic effects (CPE) in the co-cultivation of CAEV and primary synovial cells. Results indicated that sensitivity, specificity were, respectively, 69.6%, 100%, for PCR; and 95.7%, 83.3% for ELISA. The PCR assay tended to have lower sensitivity and higher specificity than ELISA. When multiple tests were applied, parallel testing provided sensitivity and specificity of 98.7% and 83.3%, while series testing showed sensitivity and specificity of 66.6% and 100% respectively. These results indicated that combination of ELISA and PCR provided some advantages and possibly offered optimal methods to detect CAEV-infected goats. Kappa value of the agreement between PCR and ELISA test was 0.34, indicating fair agreement. Regarding the possibility of antigenic variation between CAEV strains used in both PCR and ELISA assays, the actual circulating CAEV strain should be reviewed in order to develop and enhance the diagnostic tests using the CAE viral antigens derived from specific local strains of Thailand.
[Mh] Termos MeSH primário: Vírus da Artrite-Encefalite Caprina
Ensaio de Imunoadsorção Enzimática/veterinária
Doenças das Cabras/virologia
Infecções por Lentivirus/veterinária
Reação em Cadeia da Polimerase/veterinária
Cultura de Vírus/veterinária
[Mh] Termos MeSH secundário: Animais
Feminino
Doenças das Cabras/diagnóstico
Cabras
Infecções por Lentivirus/diagnóstico
Infecções por Lentivirus/virologia
Reação em Cadeia da Polimerase/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171106
[Lr] Data última revisão:
171106
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170903
[St] Status:MEDLINE


  6 / 10500 MEDLINE  
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[PMID]:28818800
[Au] Autor:Johnson SA; Walsh A; Brown MR; Lute SC; Roush DJ; Burnham MS; Brorson KA
[Ad] Endereço:DBRRII, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, MD, 20993, USA. Electronic address: sarah.johnson1@fda.hhs.gov.
[Ti] Título:The step-wise framework to design a chromatography-based hydrophobicity assay for viral particles.
[So] Source:J Chromatogr B Analyt Technol Biomed Life Sci;1061-1062:430-437, 2017 Sep 01.
[Is] ISSN:1873-376X
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:A high-salt, hydrophobic interaction chromatography (HIC) method was developed to measure the relative hydrophobicity of a diverse set of solutes. Through the careful control of buffer pH and salt concentration, this assay was then used to ascertain for the first time the relative hydrophobicity values of three different bacteriophage, four mammalian viruses, and a range of biotech medicinal proteins as benchmarked to protein standards previously characterized for hydrophobicity.
[Mh] Termos MeSH primário: Cromatografia Líquida/métodos
Vírion/isolamento & purificação
[Mh] Termos MeSH secundário: Sulfato de Amônio/química
Biotecnologia
Citratos/química
Concentração de Íons de Hidrogênio
Interações Hidrofóbicas e Hidrofílicas
Cultura de Vírus
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Citrates); 1Q73Q2JULR (sodium citrate); SU46BAM238 (Ammonium Sulfate)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170906
[Lr] Data última revisão:
170906
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170819
[St] Status:MEDLINE


  7 / 10500 MEDLINE  
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[PMID]:28815386
[Au] Autor:Mor SK; Phelps NBD; Ng TFF; Subramaniam K; Primus A; Armien AG; McCann R; Puzach C; Waltzek TB; Goyal SM
[Ad] Endereço:Minnesota Veterinary Diagnostic Laboratory, Department of Veterinary Population Medicine, University of Minnesota, 1333 Gortner Avenue, St. Paul, MN, 55108, USA. kumars@umn.edu.
[Ti] Título:Genomic characterization of a novel calicivirus, FHMCV-2012, from baitfish in the USA.
[So] Source:Arch Virol;162(12):3619-3627, 2017 Dec.
[Is] ISSN:1432-8798
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:During regulatory sampling of fathead minnows (Pimephales promelas), a novel calicivirus was isolated from homogenates of kidney and spleen inoculated into bluegill fry (BF-2) cells. Infected cell cultures exhibiting cytopathic effects were screened by PCR-based methods for selected fish viral pathogens. Illumina HiSeq next generation sequencing of the total RNA revealed a novel calicivirus genome that showed limited protein sequence similarity to known homologs in a BLASTp search. The complete genome of this fathead minnow calicivirus (FHMCV) is 6564 nt long, encoding a polyprotein of 2114 aa in length. The complete polyprotein shared only 21% identity with Atlantic salmon calicivirus,followed by 11% to 14% identity with mammalian caliciviruses. A molecular detection assay (RT-PCR) was designed from this sequence for screening of field samples for FHMCV in the future. This virus likely represents a prototype species of a novel genus in the family Caliciviridae, tentatively named "Minovirus".
[Mh] Termos MeSH primário: Infecções por Caliciviridae/veterinária
Caliciviridae/classificação
Caliciviridae/isolamento & purificação
Cyprinidae/virologia
Genoma Viral
Filogenia
[Mh] Termos MeSH secundário: Estruturas Animais/virologia
Animais
Caliciviridae/genética
Infecções por Caliciviridae/virologia
Células Cultivadas
Efeito Citopatogênico Viral
Genômica
Rim/virologia
Reação em Cadeia da Polimerase
RNA Viral/genética
Análise de Sequência de DNA
Homologia de Sequência
Baço/virologia
Estados Unidos
Proteínas Virais/genética
Cultura de Vírus
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral); 0 (Viral Proteins)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171113
[Lr] Data última revisão:
171113
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170818
[St] Status:MEDLINE
[do] DOI:10.1007/s00705-017-3519-6


  8 / 10500 MEDLINE  
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[PMID]:28777951
[Au] Autor:Ghietto LM; Toigo D'Angelo AP; Viale FA; Adamo MP
[Ad] Endereço:Instituto de Virología "Dr. J. M. Vanella", Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Argentina.
[Ti] Título:Human bocavirus 1 infection of CACO-2 cell line cultures.
[So] Source:Virology;510:273-280, 2017 Oct.
[Is] ISSN:1096-0341
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Human bocavirus 1 (HBoV1) is a parvovirus associated with pneumonia in infants. It has been detected in different tissues, including colorectal tumors. In this study, we investigated whether Caco-2 cell line, derived from human colon cancer, can be utilized as a model for HBoV1 replication. We demonstrate HBoV1 replication in Caco-2 cultures supplemented with DEAE-dextran after inoculation with respiratory material from infected patients presenting with acute respiratory infection. A viral cycle of rapid development is displayed. However, in spite of HBoV1 DNA 4-fold increment in the supernatants and monolayers by day 1, evidencing that the system allows the virus genome replication after the entry occurred, infectious progeny particles were not produced. These results are consistent with an infection that is limited to a single growth cycle, which can be associated to mutations in the NS1 and VP1/VP2 regions of HBoV1 genome. Further research will contribute to fully elucidate these observations.
[Mh] Termos MeSH primário: Células Epiteliais/virologia
Bocavirus Humano/fisiologia
Cultura de Vírus
Replicação Viral
[Mh] Termos MeSH secundário: Células CACO-2
DNA Viral/análise
Seres Humanos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Viral)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170829
[Lr] Data última revisão:
170829
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170805
[St] Status:MEDLINE


  9 / 10500 MEDLINE  
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[PMID]:28576653
[Au] Autor:Cêtre-Sossah C; Dickmu S; Kwiatek O; Albina E
[Ad] Endereço:CIRAD UMR ASTRE, F-97490 Sainte Clotilde, France; INRA, UMR1309 CMAEE, F-34398 Montpellier, France. Electronic address: catherine.cetre-sossah@cirad.fr.
[Ti] Título:A G-protein-coupled chemokine receptor: A putative insertion site for a multi-pathogen recombinant capripoxvirus vaccine strategy.
[So] Source:J Immunol Methods;448:112-115, 2017 Sep.
[Is] ISSN:1872-7905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Capripoxviruses (CaPVs) have been shown to be ideal viral vectors for the development of recombinant multivalent vaccines to enable delivery of immunogenic genes from ruminant pathogens. So far, the viral thymidine kinase (TK) gene is the only gene used to generate recombinants. A putative non-essential gene encoding a G-protein-coupled chemokine receptor subfamily homologue (GPCR) was targeted as an additional insertion site. Peste des petits ruminants (PPR) was chosen as a disease model. A new recombinant CaPV expressing the viral attachment hemagglutinin (H) of the PPR virus (PPRV) in the GPCR insertion site (rKS1-HPPR-GPCR) was generated in the backbone North African isolate KS1 strain of lumpy skin disease virus (LSDV). Comparison with the recombinant CaPV expressing the H of PPRV in the TK gene (rKS1-HPPR-TK) shown to induce protection against both PPR and LSD in both sheep and goats was assessed. The suitability of the GPCR gene to be a putative additional insertion site in the CaPV genome is evaluated and discussed.
[Mh] Termos MeSH primário: Capripoxvirus/genética
Vetores Genéticos
Doença Nodular Cutânea/prevenção & controle
Vírus da Doença Nodular Cutânea/genética
Mutagênese Insercional
Peste dos Pequenos Ruminantes/prevenção & controle
Vírus da Peste dos Pequenos Ruminantes/genética
Receptores de Quimiocinas/genética
Receptores Acoplados a Proteínas-G/genética
Vacinas Virais/genética
[Mh] Termos MeSH secundário: Animais
Anticorpos Antivirais/sangue
Bovinos
Cercopithecus aethiops
Cabras
Hemaglutininas Virais/administração & dosagem
Hemaglutininas Virais/genética
Hemaglutininas Virais/imunologia
Injeções Subcutâneas
Doença Nodular Cutânea/genética
Doença Nodular Cutânea/imunologia
Doença Nodular Cutânea/virologia
Vírus da Doença Nodular Cutânea/imunologia
Peste dos Pequenos Ruminantes/genética
Peste dos Pequenos Ruminantes/imunologia
Peste dos Pequenos Ruminantes/virologia
Vírus da Peste dos Pequenos Ruminantes/imunologia
Receptores de Quimiocinas/administração & dosagem
Receptores de Quimiocinas/imunologia
Receptores Acoplados a Proteínas-G/administração & dosagem
Receptores Acoplados a Proteínas-G/imunologia
Vacinas Sintéticas/administração & dosagem
Vacinas Sintéticas/genética
Vacinas Sintéticas/imunologia
Células Vero
Vacinas Virais/administração & dosagem
Vacinas Virais/imunologia
Cultura de Vírus
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Viral); 0 (Hemagglutinins, Viral); 0 (Receptors, Chemokine); 0 (Receptors, G-Protein-Coupled); 0 (Vaccines, Synthetic); 0 (Viral Vaccines)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170918
[Lr] Data última revisão:
170918
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170604
[St] Status:MEDLINE


  10 / 10500 MEDLINE  
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[PMID]:28539437
[Au] Autor:Ruedas JB; Ladner JT; Ettinger CR; Gummuluru S; Palacios G; Connor JH
[Ad] Endereço:Department of Microbiology, Boston University School of Medicine, Boston, Massachusetts, USA jruedas@bu.edu jhconnor@bu.edu.
[Ti] Título:Spontaneous Mutation at Amino Acid 544 of the Ebola Virus Glycoprotein Potentiates Virus Entry and Selection in Tissue Culture.
[So] Source:J Virol;91(15), 2017 Aug 01.
[Is] ISSN:1098-5514
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Ebolaviruses have a surface glycoprotein (GP ) that is required for virus attachment and entry into cells. Mutations affecting GP functions can alter virus growth properties. We generated a recombinant vesicular stomatitis virus encoding Ebola virus Makona variant GP (rVSV-MAK-GP) and observed emergence of a T544I mutation in the Makona GP gene during tissue culture passage in certain cell lines. The T544I mutation emerged within two passages when VSV-MAK-GP was grown on Vero E6, Vero, and BS-C-1 cells but not when it was passaged on Huh7 and HepG2 cells. The mutation led to a marked increase in virus growth kinetics and conferred a robust growth advantage over wild-type rVSV-MAK-GP on Vero E6 cells. Analysis of complete viral genomes collected from patients in western Africa indicated that this mutation was not found in Ebola virus clinical samples. However, we observed the emergence of T544I during serial passage of various Ebola Makona isolates on Vero E6 cells. Three independent isolates showed emergence of T544I from undetectable levels in nonpassaged virus or virus passaged once to frequencies of greater than 60% within a single passage, consistent with it being a tissue culture adaptation. Intriguingly, T544I is not found in any Sudan, Bundibugyo, or Tai Forest ebolavirus sequences. Furthermore, T544I did not emerge when we serially passaged recombinant VSV encoding GP from these ebolaviruses. This report provides experimental evidence that the spontaneous mutation T544I is a tissue culture adaptation in certain cell lines and that it may be unique for the species The Ebola virus (Zaire) species is the most lethal species of all ebolaviruses in terms of mortality rate and number of deaths. Understanding how the Ebola virus surface glycoprotein functions to facilitate entry in cells is an area of intense research. Recently, three groups independently identified a polymorphism in the Ebola glycoprotein (I544) that enhanced virus entry, but they did not agree in their conclusions regarding its impact on pathogenesis. Our findings here address the origins of this polymorphism and provide experimental evidence showing that it is the result of a spontaneous mutation (T544I) specific to tissue culture conditions, suggesting that it has no role in pathogenesis. We further show that this mutation may be unique to the species , as it does not occur in Sudan, Bundibugyo, and Tai Forest ebolaviruses. Understanding the mechanism behind this mutation can provide insight into functional differences that exist in culture conditions and among ebolavirus glycoproteins.
[Mh] Termos MeSH primário: Ebolavirus/fisiologia
Proteínas Mutantes/genética
Mutação de Sentido Incorreto
Seleção Genética
Proteínas do Envelope Viral/genética
Internalização do Vírus
[Mh] Termos MeSH secundário: Adaptação Biológica
Substituição de Aminoácidos
Animais
Linhagem Celular
Análise Mutacional de DNA
Ebolavirus/genética
Ebolavirus/crescimento & desenvolvimento
Genoma Viral
Seres Humanos
Recombinação Genética
Genética Reversa
Análise de Sequência de DNA
Inoculações Seriadas
Vesiculovirus/genética
Vesiculovirus/crescimento & desenvolvimento
Cultura de Vírus
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Mutant Proteins); 0 (Viral Envelope Proteins); 0 (envelope glycoprotein, Ebola virus)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171024
[Lr] Data última revisão:
171024
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170526
[St] Status:MEDLINE



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