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Phenotypic Analysis of Yellow Fever Virus Derived from Complementary DNA

R. S. MarchevskyFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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J. MarianoFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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V. S. FerreiraFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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E. AlmeidaFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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M. J. CerqueiraFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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R. CarvalhoFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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J. W. PissurnoFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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A. P. A. Travassos Da RosaFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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M. C. SimoesFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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C. N. D. SantosFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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I. I. FerreiraFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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I. R. MuylaertFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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G. F. MannFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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C. M. RiceFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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R. GallerFundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiologicos, Departamento de Bioquimica e Biologia Molecular, Department of Molecular Microbiology, Washington University School of Medicine, Instituto Evandro Chagas, Secao de Arbovirus, Rio de Janeiro, Brazil

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A thorough phenotypic characterization of yellow fever (YF) virus generated from cDNA is a necessary prerequisite for mapping virulence/attenuation determinants and exploring the potential of YF attenuated virus 17D as a carrier for heterologous protective epitopes. In this study, YF virus was produced from 17D cDNA clones after lipofectin-mediated RNA transfection of certified primary cultures of chicken embryo fibroblasts (YFiv5.2/SL). This virus was passaged once in embryonated chicken eggs according to current YF vaccine manufacture methodology to produce the experimental virus (YFiv5.2/VL). These viruses were characterized in established monkey neurovirulence safety tests and quantitative clinical and histologic scores were derived for each virus. The experimental vaccine viruses (YFiv5.2/SL and VL) compared favorably with another well-known YF vaccine strain (17DD) used as control virus for the histologic score. Although slightly higher clinical neurovirulence was observed for YFiv5.2 as compared with the 17DD virus, it should not preclude the use of YFiv5.2 for mapping YF virus virulence determinants.

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