{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T00:56:03Z","timestamp":1776128163923,"version":"3.50.1"},"reference-count":44,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T00:00:00Z","timestamp":1764201600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>ZIKA virus (ZIKV) infections in human neonates and adults are associated with deleterious effects on brain cognition and neurological disorders. The mechanism(s) of ZIKV infection in neurons and associated neuronal antiviral responses are not fully understood. In this study, we determined the effects of ZIKV infectivity in human neuronal (SH-SY5Y) cells and mouse N2a cells\/primary cultures of murine cortical neurons at early and late tested timepoints of infection. The human neuronal cells had higher ZIKV loads compared to the mouse N2a cells, but the viral loads in the murine cortical neurons were between the loads in these two in vitro cell lines. The murine cortical neurons were thought to be more permissive to ZIKV infection, but viral infection kinetics showed a declining trend like that observed in the mouse N2a cells. We noted that infectious extracellular vesicle (EV)-mediated ZIKV infection showed higher viral loads in the SH-SY5Y cells compared to direct infection with laboratory virus stocks. Similar results were obtained with ZIKV infectious EVs in the mouse N2a cells and cortical neurons. In addition, we noted that ZIKV infection significantly induced EV secretion from all three neuronal cells. Also, we found that ZIKV infection modulates the expression of type 1 interferons (IFNs) and entry receptors such as Tyro3, Axl, and MER-TK (TAM). Alongside the increased ZIKV loads in the SH-SY5Y cells, IFN-beta transcript levels and receptors Tyro3\/MER-TK were upregulated at early timepoints of infection. Overall, the reduced ZIKV loads and decreasing IFN expression in the mouse neuronal cells suggested a unique murine cellular ability to restrict and limit viral replication. This could be one of the reasons for the unavailability of wild-type mouse models for ZIKV infection. Our data further shows that ZIKV may preferentially infect human rather than murine neuronal cells, and this could be the potential reason for microcephaly in newborns.<\/jats:p>","DOI":"10.3390\/ijms262311500","type":"journal-article","created":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T16:31:52Z","timestamp":1764261112000},"page":"11500","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Zika Virus Infection Is More Highly Replicative and Transmissible by Extracellular Vesicles in Human than in Mouse Neuronal Cells"],"prefix":"10.3390","volume":"26","author":[{"given":"Kehinde Damilare","family":"Fasae","sequence":"first","affiliation":[{"name":"Department of Biomedical and Diagnostic Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8796-0319","authenticated-orcid":false,"given":"Md","family":"Bayzid","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Diagnostic Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Girish","family":"Neelakanta","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Diagnostic Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6005-3089","authenticated-orcid":false,"given":"Hameeda","family":"Sultana","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Diagnostic Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bhat, E.A., Ali, T., Sajjad, N., Kumar, R., and Bron, P. (2023). Insights into the structure, functional perspective, and pathogenesis of ZIKV: An updated review. Biomed. Pharmacother., 165.","DOI":"10.1016\/j.biopha.2023.115175"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"B\u00fcttner, C., Heer, M., Traichel, J., Schwemmle, M., and Heimrich, B. (2019). Zika Virus-Mediated Death of Hippocampal Neurons Is Independent from Maturation State. Front. Cell. Neurosci., 13.","DOI":"10.3389\/fncel.2019.00389"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Komarasamy, T.V., Adnan, N.A.A., James, W., and Balasubramaniam, V.R.M.T. (2022). Zika Virus Neuropathogenesis: The Different Brain Cells, Host Factors and Mechanisms Involved. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.773191"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1126","DOI":"10.1016\/j.amjmed.2016.04.034","article-title":"The Clinical Spectrum of Zika Virus in Returning Travelers","volume":"129","author":"Meltzer","year":"2016","journal-title":"Am. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1016\/j.stem.2016.11.014","article-title":"Advances in Zika Virus Research: Stem Cell Models, Challenges, and Opportunities","volume":"19","author":"Ming","year":"2016","journal-title":"Cell Stem Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1101\/gad.298216.117","article-title":"How does Zika virus cause microcephaly?","volume":"31","author":"Wen","year":"2017","journal-title":"Genes Dev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1038\/nn.4612","article-title":"Zika virus directly infects peripheral neurons and induces cell death","volume":"20","author":"Oh","year":"2017","journal-title":"Nat. Neurosci."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Bobermin, L.D., Quincozes-Santos, A., Santos, C.L., Varela, A.P.M., Teixeira, T.F., Wartchow, K.M., Lissner, L.J., da Silva, A., Thomaz, N.K., and Santi, L. (2020). Zika virus exposure affects neuron-glia communication in the hippocampal slices of adult rats. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-78735-y"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Ledur, P.F., Karmirian, K., Pedrosa, C.d.S.G., Souza, L.R.Q., Assis-De-Lemos, G., Martins, T.M., Ferreira, J.d.C.C.G., Reis, G.F.d.A., Silva, E.S., and Silva, D. (2020). Zika virus infection leads to mitochondrial failure, oxidative stress and DNA damage in human iPSC-derived astrocytes. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-57914-x"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1080\/22221751.2019.1637283","article-title":"Zika virus differentially infects human neural progenitor cells according to their state of differentiation and dysregulates neurogenesis through the Notch pathway","volume":"8","author":"Ferraris","year":"2019","journal-title":"Emerg. Microbes Infect."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Mart\u00edn, C.S.-S., Li, T., Bouquet, J., Streithorst, J., Yu, G., Paranjpe, A., and Chiu, C.Y. (2018). Differentiation enhances Zika virus infection of neuronal brain cells. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-32400-7"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.meegid.2019.01.018","article-title":"ZIKA virus entry mechanisms in human cells","volume":"69","author":"Agrelli","year":"2019","journal-title":"Infect. Genet. Evol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s12987-018-0100-y","article-title":"Zika virus crosses an in vitro human blood brain barrier model","volume":"15","author":"Alimonti","year":"2018","journal-title":"Fluids Barriers CNS"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Balint, E., Montemarano, A., Feng, E., and Ashkar, A.A. (2021). From Mosquito Bites to Sexual Transmission: Evaluating Mouse Models of Zika Virus Infection. Viruses, 13.","DOI":"10.3390\/v13112244"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Nazerai, L., Sch\u00f8ller, A.S., Rasmussen, P.O.S., Buus, S., Stryhn, A., Christensen, J.P., and Thomsen, A.R. (2018). A New In Vivo Model to Study Protective Immunity to Zika Virus Infection in Mice with Intact Type I Interferon Signaling. Front. Immunol., 9.","DOI":"10.3389\/fimmu.2018.00593"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.1099\/jgv.0.001153","article-title":"In vitro and in vivo models for studying Zika virus biology","volume":"99","author":"Pena","year":"2018","journal-title":"J. Gen. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1016\/j.stem.2016.06.009","article-title":"Modeling Zika Virus Infection in Mice","volume":"19","author":"Rossi","year":"2016","journal-title":"Cell Stem Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/j.stem.2016.08.005","article-title":"Zika Virus Infects Neural Progenitors in the Adult Mouse Brain and Alters Proliferation","volume":"19","author":"Li","year":"2016","journal-title":"Cell Stem Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"247","DOI":"10.20517\/evcna.2022.30","article-title":"Alterations in arthropod and neuronal exosomes reduce virus transmission and replication in recipient cells","volume":"3","author":"Fasae","year":"2022","journal-title":"Extracell. Vesicles Circ. Nucleic Acids"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Regmi, P., Khanal, S., Neelakanta, G., and Sultana, H. (2020). Tick-Borne Flavivirus Inhibits Sphingomyelinase (IsSMase), a Venomous Spider Ortholog to Increase Sphingomyelin Lipid Levels for Its Survival in Ixodes scapularis Ticks. Front. Cell. Infect. Microbiol., 10.","DOI":"10.3389\/fcimb.2020.00244"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"E6604","DOI":"10.1073\/pnas.1720125115","article-title":"Arthropod EVs mediate dengue virus transmission through interaction with a tetraspanin domain containing glycoprotein Tsp29Fb","volume":"115","author":"Vora","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhou, W., Tahir, F., Wang, J.C.-Y., Woodson, M., Sherman, M.B., Karim, S., Neelakanta, G., and Sultana, H. (2020). Discovery of Exosomes from Tick Saliva and Salivary Glands Reveals Therapeutic Roles for CXCL12 and IL-8 in Wound Healing at the Tick\u2013Human Skin Interface. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.00554"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zhou, W., Woodson, M., Neupane, B., Bai, F., Sherman, M.B., Choi, K.H., Neelakanta, G., and Sultana, H. (2018). Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells. PLoS Pathog., 14.","DOI":"10.1371\/journal.ppat.1006764"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1080\/22221751.2019.1578188","article-title":"Exosomes mediate Zika virus transmission through SMPD3 neutral Sphingomyelinase in cortical neurons","volume":"8","author":"Zhou","year":"2019","journal-title":"Emerg. Microbes Infect."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2007","DOI":"10.1038\/s41467-023-37698-0","article-title":"Type I interferon shapes brain distribution and tropism of tick-borne flavivirus","volume":"14","author":"Chotiwan","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Lee, I., Bos, S., Li, G., Wang, S., Gadea, G., Despr\u00e8s, P., and Zhao, R.Y. (2018). Probing Molecular Insights into Zika Virus\u2013Host Interactions. Viruses, 10.","DOI":"10.20944\/preprints201804.0063.v1"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/j.virol.2018.03.009","article-title":"Infection by Zika viruses requires the transmembrane protein AXL, endocytosis and low pH","volume":"518","author":"Persaud","year":"2018","journal-title":"Virology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/j.isci.2019.03.003","article-title":"Loss of the TAM Receptor Axl Ameliorates Severe Zika Virus Pathogenesis and Reduces Apoptosis in Microglia","volume":"13","author":"Hastings","year":"2019","journal-title":"iScience"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1016\/j.celrep.2017.03.058","article-title":"TAM Receptors Are Not Required for Zika Virus Infection in Mice","volume":"19","author":"Hastings","year":"2017","journal-title":"Cell Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1093\/infdis\/jix511","article-title":"Neuroviruses Emerging in the Americas Study Neurological Implications of Zika Virus Infection in Adults","volume":"216","author":"Parra","year":"2017","journal-title":"J. Infect. Dis."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"114469","DOI":"10.1016\/j.expneurol.2023.114469","article-title":"Neurotoxic properties of the Zika virus envelope protein","volume":"367","author":"Steiner","year":"2023","journal-title":"Exp. Neurol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Dahiya, N., Yadav, M., Singh, H., Jakhar, R., and Sehrawat, N. (2023). ZIKV: Epidemiology, infection mechanism and current therapeutics. Front. Trop. Dis., 3.","DOI":"10.3389\/fitd.2022.1059283"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1016\/j.cell.2016.09.020","article-title":"Zika Virus: Immunity and Vaccine Development","volume":"167","author":"Pierson","year":"2016","journal-title":"Cell"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.antiviral.2017.07.010","article-title":"Replication of the Zika virus in different iPSC-derived neuronal cells and implications to assess efficacy of antivirals","volume":"145","author":"Lanko","year":"2017","journal-title":"Antivir. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3890","DOI":"10.1038\/s41467-019-11866-7","article-title":"Zika virus replicates in adult human brain tissue and impairs synapses and memory in mice","volume":"10","author":"Figueiredo","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1186\/s13287-020-02033-7","article-title":"Detection of viral RNA fragments in human iPSC cardiomyocytes following treatment with extracellular vesicles from SARS-CoV-2 coding sequence overexpressing lung epithelial cells","volume":"11","author":"Kwon","year":"2020","journal-title":"Stem Cell Res. Ther."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Rey-Cadilhac, F., Rachenne, F., Miss\u00e9, D., and Pompon, J. (2023). Viral Components Trafficking with(in) Extracellular Vesicles. Viruses, 15.","DOI":"10.3390\/v15122333"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1120","DOI":"10.1002\/eji.201847483","article-title":"Infection with a Brazilian isolate of Zika virus generates RIG-I stimulatory RNA and the viral NS5 protein blocks type I IFN induction and signaling","volume":"48","author":"Hertzog","year":"2018","journal-title":"Eur. J. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.virol.2018.11.009","article-title":"Zika virus NS5 protein antagonizes type I interferon production via blocking TBK1 activation","volume":"527","author":"Lin","year":"2019","journal-title":"Virology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"17006","DOI":"10.1038\/celldisc.2017.6","article-title":"Zika virus evades interferon-mediated antiviral response through the co-operation of multiple nonstructural proteins in vitro","volume":"3","author":"Wu","year":"2017","journal-title":"Cell Discov."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1016\/j.chom.2016.03.010","article-title":"A Mouse Model of Zika Virus Pathogenesis","volume":"19","author":"Lazear","year":"2016","journal-title":"Cell Host Microbe"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3150","DOI":"10.4049\/jimmunol.1201140","article-title":"Semaphorin 7A contributes to West Nile virus pathogenesis through TGF-\u03b21\/Smad6 signaling","volume":"189","author":"Sultana","year":"2012","journal-title":"J. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Bhowmick, B., Ahmed, W., Neelakanta, G., and Sultana, H. (2025). Pharmacological Agent GW4869 Inhibits Tick-Borne Langat Virus Replication to Affect Extracellular Vesicles Secretion. Viruses, 17.","DOI":"10.3390\/v17070969"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"110391","DOI":"10.1016\/j.isci.2024.110391","article-title":"GW4869 inhibitor affects vector competence and tick-borne flavivirus acquisition and transmission by blocking exosome secretion","volume":"27","author":"Sultana","year":"2024","journal-title":"iScience"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/23\/11500\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T16:32:21Z","timestamp":1764261141000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/23\/11500"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,27]]},"references-count":44,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["ijms262311500"],"URL":"https:\/\/doi.org\/10.3390\/ijms262311500","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,27]]}}}