title: Production of yellow fever 17DD vaccine virus in primary culture of chicken embryo fibroblasts: yields, thermo and genetic stability, attenuation and immunogenicity.
abstract: While a good vaccine against yellow fever (YF) virus has been available for decades, the basic technology for the production of YF vaccine in chicken embryos has remained substantially unchanged since the 1940s. Here we describe the highly efficient and economic production of the 17DD strain of YF virus in chicken embryo fibroblast (CEF) cell cultures with viral titers ranging from 6.3 to 6.7 log10PFU/mL. Thermostability of two different formulations (5 and 50-dose vials) of the CEF vaccine virus was found to be as high as the current vaccines retaining the minimal titer required for YF 17D vaccines. The production passage in CEF did not lead to the selection of genetic variants as shown by nucleotide sequence analyses of the CEF-derived vaccine lots or the sequence of viruses recovered from monkeys experimentally inoculated with the CEF virus. YF 17DD virus produced in CEF was also indistinguishable from its seed lot virus parent in terms of plaque size and immunogenicity in mice and monkeys. Comparison of the CEF virus and the seed lot virus made in chicken embryo in the internationally accepted monkey neurovirulence test (MNVT) revealed a higher clinical score for the former. The differences in central nervous system (CNS) histological scores for monkeys inoculated with the chicken embryo and experimental CEF vaccines were at the borderline level of statistical significance. These data warrant further studies on the monkey attenuation of other batches of CEF-derived vaccines.
authors.affiliation: Fundacao Oswaldo Cruz, Instituto de Tecnologia em Imunobiológicos, Avenida Brasil 4365, Manguinhos, Rio de Janeiro 21045-900, Brazil.
authors.forename: Marcos S
authors.initials: MS
authors.lastname: Freire
authors.forename: George F
authors.initials: GF
authors.lastname: Mann
authors.forename: Renato S
authors.initials: RS
authors.lastname: Marchevsky
authors.forename: Anna M Y
authors.initials: AM
authors.lastname: Yamamura
authors.forename: Luiz F C
authors.initials: LF
authors.lastname: Almeida
authors.forename: Alfredo V
authors.initials: AV
authors.lastname: Jabor
authors.forename: José M N
authors.initials: JM
authors.lastname: Malachias
authors.forename: Evandro S F
authors.initials: ES
authors.lastname: Coutinho
authors.forename: Ricardo
authors.initials: R
authors.lastname: Galler
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D005260:Female; D005347:Fibroblasts / Q000821:virology*; D008253:Macaca mulatta; D008297:Male; D051379:Mice; D023421:Models, Animal; D009154:Mutation / Q000235:genetics; D013696:Temperature; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology; D014709:Vero Cells; D010948:Viral Plaque Assay; D014774:Virulence; D014776:Virus Cultivation; D015004:Yellow Fever / Q000517:prevention & control; D022341:Yellow Fever Vaccine / Q000235:genetics / Q000276:immunology; D015005:Yellow fever virus / Q000235:genetics / Q000254:growth & development* / Q000276:immunology
chemical_list: D014613:Vaccines, Attenuated; D022341:Yellow Fever Vaccine