{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T03:22:09Z","timestamp":1782789729501,"version":"3.54.5"},"reference-count":78,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2019,6,17]],"date-time":"2019-06-17T00:00:00Z","timestamp":1560729600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008393","name":"Teknologi og Produktion, Det Frie Forskningsr\u00e5d","doi-asserted-by":"publisher","award":["6111-00104B"],"award-info":[{"award-number":["6111-00104B"]}],"id":[{"id":"10.13039\/100008393","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Chronic viral disease constitutes a major global health problem, with several hundred million people affected and an associated elevated number of deaths. An increasing number of disorders caused by human flaviviruses are related to their capacity to establish a persistent infection. Here we show that Usutu virus (USUV), an emerging zoonotic flavivirus linked to sporadic neurologic disease in humans, can establish a persistent infection in cell culture. Two independent lineages of Vero cells surviving USUV lytic infection were cultured over 82 days (41 cell transfers) without any apparent cytopathology crisis associated. We found elevated titers in the supernatant of these cells, with modest fluctuations during passages but no overall tendency towards increased or decreased infectivity. In addition to full-length genomes, viral RNA isolated from these cells at passage 40 revealed the presence of defective genomes, containing different deletions at the 5\u2019 end. These truncated transcripts were all predicted to encode shorter polyprotein products lacking membrane and envelope structural proteins, and most of non-structural protein 1. Treatment with different broad-range antiviral nucleosides revealed that USUV is sensitive to these compounds in the context of a persistent infection, in agreement with previous observations during lytic infections. The exposure of infected cells to prolonged treatment (10 days) with favipiravir and\/or ribavirin resulted in the complete clearance of infectivity in the cellular supernatants (decrease of ~5 log10 in virus titers and RNA levels), although modest changes in intracellular viral RNA levels were recorded (&lt;2 log10 decrease). Drug withdrawal after treatment day 10 resulted in a relapse in virus titers. These results encourage the use of persistently-infected cultures as a surrogate system in the identification of improved antivirals against flaviviral chronic disease.<\/jats:p>","DOI":"10.3390\/v11060560","type":"journal-article","created":{"date-parts":[[2019,6,17]],"date-time":"2019-06-17T11:24:45Z","timestamp":1560770685000},"page":"560","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Establishment of a Cell Culture Model of Persistent Flaviviral Infection: Usutu Virus Shows Sustained Replication during Passages and Resistance to Extinction by Antiviral Nucleosides"],"prefix":"10.3390","volume":"11","author":[{"given":"Raquel Navarro","family":"Sempere","sequence":"first","affiliation":[{"name":"Life Science &amp; Bioengineering Building, Technical University of Denmark, 2800 Kongens Lyngby, Denmark"},{"name":"Abiopep Sociedad Limitada, Parque Cient\u00edfico de Murcia, 30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4138-4608","authenticated-orcid":false,"given":"Armando","family":"Arias","sequence":"additional","affiliation":[{"name":"Life Science &amp; Bioengineering Building, Technical University of Denmark, 2800 Kongens Lyngby, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,6,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.cell.2009.06.036","article-title":"Redefining chronic viral infection","volume":"138","author":"Virgin","year":"2009","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2194","DOI":"10.4254\/wjh.v7.i19.2194","article-title":"Chronic hepatitis E: A brief review","volume":"7","author":"Murali","year":"2015","journal-title":"World J. Hepatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2676","DOI":"10.4254\/wjh.v7.i26.2676","article-title":"Hepatitis C virus: A global view","volume":"7","author":"Mohamed","year":"2015","journal-title":"World J. Hepatol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1111\/cei.12884","article-title":"Chronic norovirus infection and common variable immunodeficiency","volume":"188","author":"Woodward","year":"2017","journal-title":"Clin. Exp. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Capizzi, T., Makari-Judson, G., Steingart, R., and Mertens, W.C. (2011). Chronic diarrhea associated with persistent norovirus excretion in patients with chronic lymphocytic leukemia: Report of two cases. BMC Infect. Dis., 11.","DOI":"10.1186\/1471-2334-11-131"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1111\/2049-632X.12178","article-title":"The role of viral persistence in flavivirus biology","volume":"71","author":"Mlera","year":"2014","journal-title":"Pathog. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"493","DOI":"10.2217\/fvl-2018-0021","article-title":"Defective (interfering) viral genomes re-explored: Impact on antiviral immunity and virus persistence","volume":"13","author":"Manzoni","year":"2018","journal-title":"Future Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/S0140-6736(17)31832-9","article-title":"Ending AIDS: Myth or reality?","volume":"390","author":"Williams","year":"2017","journal-title":"Lancet"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.chom.2017.01.004","article-title":"Zika Virus Pathogenesis and Tissue Tropism","volume":"21","author":"Miner","year":"2017","journal-title":"Cell Host Microbe"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5914","DOI":"10.4049\/jimmunol.0900255","article-title":"Persistent Chronic Inflammation and Infection by Chikungunya Arthritogenic Alphavirus in Spite of a Robust Host Immune Response","volume":"184","author":"Hoarau","year":"2010","journal-title":"J. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1016\/S0140-6736(08)60238-X","article-title":"Pathogenic flaviviruses","volume":"371","author":"Gould","year":"2008","journal-title":"Lancet"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.micinf.2014.12.003","article-title":"Persistence of West Nile virus","volume":"17","author":"Garcia","year":"2015","journal-title":"Microbes Infect."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.4269\/ajtmh.17-0405","article-title":"Visualization of West Nile Virus in Urine Sediment using Electron Microscopy and Immunogold up to Nine Years Postinfection","volume":"97","author":"Murray","year":"2017","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Saxena, V., Xie, G., Li, B., Farris, T., Welte, T., Gong, B., Boor, P., Wu, P., Tang, S.-J., and Tesh, R. (2013). A Hamster-Derived West Nile Virus Isolate Induces Persistent Renal Infection in Mice. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0002275"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"455","DOI":"10.4269\/ajtmh.12-0110","article-title":"Pathogenesis of Modoc Virus (Flaviviridae; Flavivirus) in Persistently Infected Hamsters","volume":"88","author":"Adams","year":"2013","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1362","DOI":"10.1128\/iai.10.6.1362-1369.1974","article-title":"Modoc viral infections in the deer mouse Peromyscus maniculatus","volume":"10","author":"Davis","year":"1974","journal-title":"Infect. Immun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1128\/jvi.39.2.413-421.1981","article-title":"Isolation of a replication-efficient mutant of West Nile virus from a persistently infected genetically resistant mouse cell culture","volume":"39","author":"Brinton","year":"1981","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.1016\/S1473-3099(16)30193-1","article-title":"Zika virus in the female genital tract","volume":"16","author":"Prisant","year":"2016","journal-title":"Lancet Infect. Dis."},{"key":"ref_19","first-page":"NEJMoa1613108","article-title":"Persistence of Zika Virus in Body Fluids\u2014Preliminary Report","volume":"379","author":"Rosenberg","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2195","DOI":"10.1056\/NEJMc1604449","article-title":"Evidence of Sexual Transmission of Zika Virus","volume":"374","author":"Matheron","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1016\/j.cell.2016.08.004","article-title":"Vaginal Exposure to Zika Virus during Pregnancy Leads to Fetal Brain Infection","volume":"166","author":"Yockey","year":"2016","journal-title":"Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1128\/JVI.77.1.25-36.2003","article-title":"Characterization of a Siberian Virus Isolated from a Patient with Progressive Chronic Tick-Borne Encephalitis","volume":"77","author":"Gritsun","year":"2003","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1002\/jmv.1890400412","article-title":"Persistence of Japanese encephalitis virus in the human nervous system","volume":"40","author":"Ravi","year":"1993","journal-title":"J. Med. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1603\/ME10280","article-title":"Relationships between infection, dissemination, and transmission of West Nile virus RNA in Culex pipiens quinquefasciatus (Diptera: Culicidae)","volume":"49","author":"Richards","year":"2012","journal-title":"J. Med. Entomol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1089\/1530366041210729","article-title":"West Nile Virus Dissemination and Tissue Tropisms in Orally Infected Culex pipiens quinquefasciatus","volume":"4","author":"Girard","year":"2004","journal-title":"Vector-Borne Zoonotic Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1016\/j.ttbdis.2017.07.008","article-title":"Properties of the tick-borne encephalitis virus population during persistent infection of ixodid ticks and tick cell lines","volume":"8","author":"Belova","year":"2017","journal-title":"Ticks Tick-Borne Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e00614-15","DOI":"10.1128\/mBio.00614-15","article-title":"Development of a Model System for Tick-Borne Flavivirus Persistence in HEK 293T Cells","volume":"6","author":"Mlera","year":"2015","journal-title":"MBio"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e00314-16","DOI":"10.1128\/mBio.00314-16","article-title":"Transcriptome Analysis Reveals a Signature Profile for Tick-Borne Flavivirus Persistence in HEK 293T Cells","volume":"7","author":"Mlera","year":"2016","journal-title":"MBio"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Offerdahl, D.K., Dorward, D.W., Hansen, B.T., and Bloom, M.E. (2012). A Three-Dimensional Comparison of Tick-Borne Flavivirus Infection in Mammalian and Tick Cell Lines. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0047912"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Mlera, L., Melik, W., Offerdahl, D., Dahlstrom, E., Porcella, S., and Bloom, M. (2016). Analysis of the Langat Virus Genome in Persistent Infection of an Ixodes scapularis Cell Line. Viruses, 8.","DOI":"10.3390\/v8090252"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1128\/JVI.02492-14","article-title":"Persistent Hepatitis C Virus Infection Impairs Ribavirin Antiviral Activity through Clathrin-Mediated Trafficking of Equilibrative Nucleoside Transporter 1","volume":"89","author":"Panigrahi","year":"2015","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1007\/978-1-4939-8976-8_10","article-title":"A Cell Culture Model for Persistent HCV Infection","volume":"Volume 1911","author":"Castro","year":"2019","journal-title":"Methods in Molecular Biology"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1016\/j.jmii.2016.11.004","article-title":"Viral dynamics of persistent hepatitis C virus infection in high-sensitive reporter cells resemble patient\u2019s viremia","volume":"51","author":"Tsai","year":"2017","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"11082","DOI":"10.1128\/JVI.01307-06","article-title":"Persistent hepatitis C virus infection in vitro: Coevolution of virus and host","volume":"80","author":"Zhong","year":"2006","journal-title":"J. Virol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"30391","DOI":"10.2807\/1560-7917.ES.2016.21.45.30391","article-title":"Widespread Usutu virus outbreak in birds in The Netherlands, 2016","volume":"21","author":"Rijks","year":"2016","journal-title":"Eurosurveillance"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"219","DOI":"10.3390\/v7010219","article-title":"Usutu virus: An emerging flavivirus in Europe","volume":"7","author":"Ashraf","year":"2015","journal-title":"Viruses"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"17","DOI":"10.2807\/1560-7917.ES.2017.22.41.17-00644","article-title":"Usutu virus infections among blood donors, Austria, July and August 2017\u2014Raising awareness for diagnostic challenges","volume":"22","author":"Bakonyi","year":"2017","journal-title":"Eurosurveillance"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1800545","DOI":"10.2807\/1560-7917.ES.2018.23.43.1800545","article-title":"Increase in human West Nile and Usutu virus infections, Austria, 2018","volume":"23","author":"Aberle","year":"2018","journal-title":"Eurosurveillance"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"30501","DOI":"10.2807\/1560-7917.ES.2017.22.14.30501","article-title":"Blood donor screening for West Nile virus (WNV) revealed acute Usutu virus (USUV) infection, Germany, September 2016","volume":"22","author":"Cadar","year":"2017","journal-title":"Eurosurveillance"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.cmi.2016.09.019","article-title":"Usutu virus infections in humans: A retrospective analysis in the municipality of Modena, Italy","volume":"23","author":"Grottola","year":"2017","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1007\/s13365-014-0300-4","article-title":"First cases of human Usutu virus neuroinvasive infection in Croatia, August\u2013September 2013: Clinical and laboratory features","volume":"21","author":"Santini","year":"2015","journal-title":"J. Neurovirol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"19446","DOI":"10.2807\/ese.14.50.19446-en","article-title":"First human case of Usutu virus neuroinvasive infection, Italy, August\u2013September 2009","volume":"14","author":"Pecorari","year":"2009","journal-title":"Eurosurveillance"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"875","DOI":"10.3201\/eid2405.171122","article-title":"Human Usutu Virus Infection with Atypical Neurologic Presentation, Montpellier, France, 2016","volume":"24","author":"Simonin","year":"2018","journal-title":"Emerg. Infect. Dis."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/j.micinf.2017.05.004","article-title":"Is Usutu virus ready for prime time?","volume":"19","author":"Tetro","year":"2017","journal-title":"Microbes Infect."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e00380-18","DOI":"10.1128\/AAC.00380-18","article-title":"Extinction of Zika virus and Usutu virus by lethal mutagenesis reveals different patterns of sensitivity to three mutagenic drugs","volume":"62","author":"Bassi","year":"2018","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.antiviral.2018.10.026","article-title":"Favipiravir inhibits in vitro Usutu virus replication and delays disease progression in an infection model in mice","volume":"160","author":"Sharma","year":"2018","journal-title":"Antivir. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"652","DOI":"10.3201\/eid0807.020094","article-title":"Emergence of Usutu virus, an African Mosquito-Borne Flavivirus of the Japanese Encephalitis Virus Group, Central Europe","volume":"8","author":"Kolodziejek","year":"2002","journal-title":"Emerg. Infect. Dis."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1007\/978-1-59745-394-3_24","article-title":"Measuring HCV Infectivity Produced in Cell Culture and In Vivo","volume":"Volume 510","author":"Lindenbach","year":"2019","journal-title":"Methods in Molecular Biology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A simple method of estimating fifty per cent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.jcv.2010.11.008","article-title":"A rapid and specific real-time RT-PCR assay to identify Usutu virus in human plasma, serum, and cerebrospinal fluid","volume":"50","author":"Cavrini","year":"2011","journal-title":"J. Clin. Virol."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Perales, C., Agudo, R., Tejero, H., Manrubia, S.C., and Domingo, E. (2009). Potential benefits of sequential inhibitor-mutagen treatments of RNA virus infections. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000658"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"e01400-17","DOI":"10.1128\/AAC.01400-17","article-title":"Extinction of west nile virus by favipiravir through lethal mutagenesis","volume":"61","author":"Domingo","year":"2017","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.antiviral.2017.12.021","article-title":"Efficacy of favipiravir (T-705) in nonhuman primates infected with Ebola virus or Marburg virus","volume":"151","author":"Bixler","year":"2018","journal-title":"Antivir. Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1080\/21645515.2017.1389363","article-title":"Are we prepared for emerging flaviviruses in Europe? Challenges for vaccination","volume":"14","author":"Kaaijk","year":"2018","journal-title":"Hum. Vaccin. Immunother."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1007\/s13365-014-0285-z","article-title":"Flaviviruses, an expanding threat in public health: Focus on dengue, West Nile, and Japanese encephalitis virus","volume":"20","author":"Daep","year":"2014","journal-title":"J. Neurovirol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/S2468-1253(16)30181-9","article-title":"Global prevalence and genotype distribution of hepatitis C virus infection in 2015: A modelling study","volume":"2","author":"Blach","year":"2017","journal-title":"Lancet Gastroenterol. Hepatol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.micpath.2016.12.018","article-title":"Characterization of non-lethal West Nile Virus (WNV) infection in horses: Subclinical pathology and innate immune response","volume":"103","author":"Hartwig","year":"2017","journal-title":"Microb. Pathog."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.virol.2015.03.020","article-title":"Limited susceptibility of mice to Usutu virus (USUV) infection and induction of flavivirus cross-protective immunity","volume":"482","author":"Petrovic","year":"2015","journal-title":"Virology"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1080\/03079450500268500","article-title":"Limited pathogenicity of Usutu virus for the domestic chicken (Gallus domesticus)","volume":"34","author":"Chvala","year":"2005","journal-title":"Avian Pathol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1007\/s00705-012-1525-2","article-title":"Detection and sequencing of defective viral genomes in C6\/36 cells persistently infected with dengue virus 2","volume":"158","year":"2013","journal-title":"Arch. Virol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"2474","DOI":"10.1128\/JVI.72.3.2474-2482.1998","article-title":"Characterization of defective viral RNA produced during persistent infection of Vero cells with Murray Valley encephalitis virus","volume":"72","author":"Lancaster","year":"1998","journal-title":"J. Virol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.virol.2012.01.028","article-title":"Internally deleted WNV genomes isolated from exotic birds in New Mexico: Function in cells, mosquitoes, and mice","volume":"427","author":"Pesko","year":"2012","journal-title":"Virology"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"11765","DOI":"10.1128\/JVI.01205-09","article-title":"Exclusion of West Nile virus superinfection through RNA replication","volume":"83","author":"Zou","year":"2009","journal-title":"J. Virol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"30452","DOI":"10.2807\/1560-7917.ES.2017.22.4.30452","article-title":"Widespread activity of multiple lineages of Usutu virus, western Europe, 2016","volume":"22","author":"Cadar","year":"2017","journal-title":"Eurosurveillance"},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Sissoko, D., Laouenan, C., Folkesson, E., M\u2019Lebing, A.-B., Beavogui, A.-H., Baize, S., Camara, A.-M., Maes, P., Shepherd, S., and Danel, C. (2016). Experimental Treatment with Favipiravir for Ebola Virus Disease (the JIKI Trial): A Historically Controlled, Single-Arm Proof-of-Concept Trial in Guinea. PLoS Med., 13.","DOI":"10.1371\/journal.pmed.1001967"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"S27","DOI":"10.1093\/infdis\/jiv280","article-title":"Advances Toward a Norovirus Antiviral: From Classical Inhibitors to Lethal Mutagenesis","volume":"213","author":"Thorne","year":"2016","journal-title":"J. Infect. Dis."},{"key":"ref_67","first-page":"323","article-title":"Antiviral Strategies Based on Lethal Mutagenesis and Error Threshold","volume":"392","author":"Perales","year":"2016","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"e03679","DOI":"10.7554\/eLife.03679","article-title":"Favipiravir elicits antiviral mutagenesis during virus replication in vivo","volume":"3","author":"Arias","year":"2014","journal-title":"Elife"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"2173","DOI":"10.1056\/NEJMc1806941","article-title":"Mutagenesis in Norovirus in Response to Favipiravir Treatment","volume":"379","author":"Ruis","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"6172","DOI":"10.1128\/JVI.02778-12","article-title":"Deep sequencing reveals mutagenic effects of ribavirin during monotherapy of hepatitis C virus genotype 1-infected patients","volume":"87","author":"Dietz","year":"2013","journal-title":"J. Virol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"3455","DOI":"10.1128\/JVI.03453-14","article-title":"Characterization of dengue virus NS4A and NS4B protein interaction","volume":"89","author":"Zou","year":"2015","journal-title":"J. Virol."},{"key":"ref_72","first-page":"266","article-title":"Stress responses in flavivirus-infected cells: Activation of unfolded protein response and autophagy","volume":"5","author":"Saiz","year":"2014","journal-title":"Front. Microbiol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"22147","DOI":"10.1074\/jbc.M110.192500","article-title":"Flavivirus NS4A-induced autophagy protects cells against death and enhances virus replication","volume":"286","author":"McLean","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"8880","DOI":"10.1128\/JVI.00354-15","article-title":"Biology of Zika Virus Infection in Human Skin Cells","volume":"89","author":"Hamel","year":"2015","journal-title":"J. Virol."},{"key":"ref_75","doi-asserted-by":"crossref","unstructured":"Bl\u00e1zquez, A.-B., Escribano-Romero, E., Merino-Ramos, T., Saiz, J.-C., and Mart\u00edn-Acebes, M.A. (2013). Infection with Usutu Virus Induces an Autophagic Response in Mammalian Cells. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0002509"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/j.stem.2016.07.019","article-title":"Zika Virus NS4A and NS4B Proteins Deregulate Akt-mTOR Signaling in Human Fetal Neural Stem Cells to Inhibit Neurogenesis and Induce Autophagy","volume":"19","author":"Liang","year":"2016","journal-title":"Cell Stem Cell"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.virusres.2017.09.006","article-title":"Role of autophagy in Zika virus infection and pathogenesis","volume":"254","author":"Chiramel","year":"2018","journal-title":"Virus Res."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"2303","DOI":"10.1084\/jem.20170957","article-title":"Inhibition of autophagy limits vertical transmission of Zika virus in pregnant mice","volume":"214","author":"Cao","year":"2017","journal-title":"J. Exp. Med."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/6\/560\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:59:00Z","timestamp":1760187540000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/6\/560"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,6,17]]},"references-count":78,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2019,6]]}},"alternative-id":["v11060560"],"URL":"https:\/\/doi.org\/10.3390\/v11060560","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,6,17]]}}}