{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T16:32:06Z","timestamp":1790440326440,"version":"4.1.0"},"reference-count":26,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,3,4]],"date-time":"2022-03-04T00:00:00Z","timestamp":1646352000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The disease yellow fever was prevented by two live attenuated vaccines, strains 17D and French neurotropic vaccine (FNV), derived by serial passage of wild-type (WT) strains Asibi and French Viscerotropic virus (FVV), respectively. Both 17D and FNV displayed decreased genetic diversity and resistance to the antiviral Ribavirin compared to their WT parental strains, which are thought to contribute to their attenuated phenotypes. Subsequent studies found that only a few passages of WT strain FVV in HeLa cells resulted in an attenuated virus. In the current study, the genome sequence of FVV following five passages in HeLa cells (FVV HeLa p5) was determined through Next Generation Sequencing (NGS) with the aim to investigate the molecular basis of viral attenuation. It was found that WT FVV and FVV HeLa p5 virus differed by five amino acid substitutions: E-D155A, E-K331R, E-I412V, NS2A-T105A, and NS4B-V98I. Surprisingly, the genetic diversity and Ribavirin resistance of the FVV HeLa p5 virus were not statistically different to WT parent FVV. These findings suggest that while FVV HeLa p5 is attenuated, this is not dependent on a high-fidelity replication complex, characterized by reduced genetic diversity or increased Ribavirin stability, as seen with FNV and 17D vaccines.<\/jats:p>","DOI":"10.3390\/v14030527","type":"journal-article","created":{"date-parts":[[2022,3,6]],"date-time":"2022-03-06T20:40:38Z","timestamp":1646599238000},"page":"527","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Genetic Diversity Does Not Contribute to Attenuation of HeLa Passaged Wild-Type Yellow Fever Virus Strain French Viscerotropic Virus"],"prefix":"10.3390","volume":"14","author":[{"given":"Ashley E.","family":"Strother","sequence":"first","affiliation":[{"name":"Department of Pathology, University of Texas Medical Branch (UTMB), Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jill K.","family":"Thompson","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Texas Medical Branch (UTMB), Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6512-363X","authenticated-orcid":false,"given":"Steven G.","family":"Widen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Texas Medical Branch (UTMB), Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3740-7448","authenticated-orcid":false,"given":"Alan D. T.","family":"Barrett","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Texas Medical Branch (UTMB), Galveston, TX 77555, USA"},{"name":"Sealy Institute for Vaccine Sciences, University of Texas Medical Branch (UTMB), Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.jcv.2014.08.030","article-title":"Yellow fever","volume":"64","author":"Monath","year":"2015","journal-title":"J. Clin. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Hamlet, A., Jean, K.K., Perea, W., Yactayo, S., Biey, J., Van Kerkhove, M., Ferguson, N., and Garske, T. (2018). The seasonal influence of climate and environment on yellow fever transmission across Africa. PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006284"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/s40409-018-0162-y","article-title":"Yellow fever in Africa and the Americas: A historical and epidemiological perspective","volume":"24","author":"Chippaux","year":"2018","journal-title":"J. Venom. Anim. Toxins Incl. Trop. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Hamrick, P.N., Aldighieri, S., Machado, G., Leonel, D.G., Vilca, L.M., Uriona, S., and Schneider, M.C. (2017). Geographic patterns and environmental factors associated with human yellow fever presence in the Americas. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0005897"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Tsegaye, M.M., Beyene, B., Ayele, W., Abebe, A., Tareke, I., Sall, A., Yactayo, S., Shibeshi, M.E., Staples, E., and Belay, D. (2018). Sero-prevalence of yellow fever and related Flavi viruses in Ethiopia: A public health perspective. BMC Public Health, 18.","DOI":"10.1186\/s12889-018-5726-9"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1084\/jem.65.6.787","article-title":"The use of yellow fever virus modified by in vitro cultivation for human immunization","volume":"65","author":"Theiler","year":"1937","journal-title":"J. Exp. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.4269\/ajtmh.1933.s1-13.1","article-title":"Studies on the Pathogenesis of Neurotropic Yellow Fever Virus in Macacus Rhesus 1","volume":"13","author":"Lloyd","year":"1933","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1093\/infdis\/jit546","article-title":"Comparison of the Live Attenuated Yellow Fever Vaccine 17D-204 Strain to Its Virulent Parental Strain Asibi by Deep Sequencing","volume":"209","author":"Beck","year":"2013","journal-title":"J. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"13408","DOI":"10.1038\/s41598-018-31085-2","article-title":"Analysis by deep sequencing of discontinued neurotropic yellow fever vaccine strains","volume":"8","author":"Beck","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e02294-19","DOI":"10.1128\/mBio.02294-19","article-title":"Attenuation of live-attenuated yellow fever 17d vaccine virus is localized to a high-fidelity replication complex","volume":"10","author":"Davis","year":"2019","journal-title":"mBio"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1093\/infdis\/113.1.9","article-title":"The growth of Asibi strain yellow fever virus in tissue cultures: I. Sensitivity and capacity of tissue cultures","volume":"113","author":"Hardy","year":"1963","journal-title":"J. Infect. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1128\/br.30.3.615-623.1966","article-title":"Attenuation of aerosolized yellow fever virus after passage in cell culture","volume":"30","author":"Hearn","year":"1966","journal-title":"Bacteriol. Rev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2301","DOI":"10.1099\/0022-1317-71-10-2301","article-title":"Attenuation of wild-type yellow fever virus by passage in HeLa cells","volume":"71","author":"Barrett","year":"1990","journal-title":"J. Gen. Virol."},{"key":"ref_14","first-page":"227","article-title":"Biological characterization of plaque-size variants of yellow fever virus in mosquitoes and mice","volume":"32","author":"Miller","year":"1988","journal-title":"Acta Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1006\/viro.1999.9873","article-title":"Molecular and Biological Changes Associated with HeLa Cell Attenuation of Wild-Type Yellow Fever Virus","volume":"261","author":"Dunster","year":"1999","journal-title":"Virology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.1099\/0022-1317-78-6-1353","article-title":"Yellow fever virus envelope protein has two discrete type-specific neutralizing epitopes","volume":"78","author":"Ryman","year":"1997","journal-title":"J. Gen. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Zhang, X., Jia, R., Shen, H., Wang, M., Yin, Z., and Cheng, A. (2017). Structures and functions of the envelope glycoprotein in flavivirus infections. Viruses, 9.","DOI":"10.3390\/v9110338"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/S0140-6736(20)32520-4","article-title":"Immunogenicity and safety of fractional doses of yellow fever vaccines: A randomised, double-blind, non-inferiority trial","volume":"397","author":"Kimathi","year":"2021","journal-title":"Lancet"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.virol.2009.09.001","article-title":"Structure of yellow fever virus envelope protein domain III","volume":"394","author":"Volk","year":"2009","journal-title":"Virology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/j.celrep.2018.12.065","article-title":"Double lock of a human neutralizing and protective monoclonal antibody targeting the yellow fever virus envelope","volume":"26","author":"Lu","year":"2019","journal-title":"Cell Rep."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Chen, S., Wu, Z., Wang, M., and Cheng, A. (2017). Innate immune evasion mediated by flaviviridae non-structural proteins. Viruses, 9.","DOI":"10.3390\/v9100291"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/0042-6822(89)90045-7","article-title":"Yellow fever virus proteins NS2A, NS213, and NS4B: Identification and partial N-terminal amino acid sequence analysis","volume":"169","author":"Chambers","year":"1989","journal-title":"Virology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1016\/j.jmb.2013.11.023","article-title":"Innate immunity to dengue virus infection and subversion of antiviral responses","volume":"426","author":"Green","year":"2013","journal-title":"J. Mol. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1002\/rmv.1835","article-title":"Flaviviral NS4b, chameleon and jack-in-the-box roles in viral replication and pathogenesis, and a molecular target for antiviral intervention","volume":"25","author":"Zmurko","year":"2015","journal-title":"Rev. Med. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1099\/vir.0.81236-0","article-title":"Genome analysis and phylogenetic relationships between east, central and west African isolates of Yellow fever virus","volume":"87","author":"Aroner","year":"2006","journal-title":"J. Gen. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1038\/s41541-021-00371-y","article-title":"Japanese encephalitis virus live attenuated vaccine strains display altered immunogenicity, virulence and genetic diversity","volume":"6","author":"Davis","year":"2021","journal-title":"NPJ Vaccines"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/3\/527\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:31:55Z","timestamp":1760135515000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/3\/527"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,4]]},"references-count":26,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["v14030527"],"URL":"https:\/\/doi.org\/10.3390\/v14030527","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,4]]}}}