{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T04:58:26Z","timestamp":1785473906421,"version":"3.56.0"},"reference-count":17,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T00:00:00Z","timestamp":1785110400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005739","name":"Universidad Nacional Aut\u00f3noma de M\u00e9xico","doi-asserted-by":"crossref","award":["IN215224"],"award-info":[{"award-number":["IN215224"]}],"id":[{"id":"10.13039\/501100005739","id-type":"DOI","asserted-by":"crossref"}]},{"award":["IN215224"],"award-info":[{"award-number":["IN215224"]}],"id":[{"id":"https:\/\/ror.org\/01tmp8f25","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003141","name":"Consejo Nacional de Ciencia y Tecnolog\u00eda","doi-asserted-by":"crossref","award":["CF-2023-I-478"],"award-info":[{"award-number":["CF-2023-I-478"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"crossref"}]},{"award":["CF-2023-I-478"],"award-info":[{"award-number":["CF-2023-I-478"]}],"id":[{"id":"https:\/\/ror.org\/00aejfc65","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020417","name":"Consejo Mexiquense de Ciencia y Tecnolog\u00eda","doi-asserted-by":"crossref","award":["FICDTEM-2023-21"],"award-info":[{"award-number":["FICDTEM-2023-21"]}],"id":[{"id":"10.13039\/501100020417","id-type":"DOI","asserted-by":"crossref"}]},{"award":["FICDTEM-2023-21"],"award-info":[{"award-number":["FICDTEM-2023-21"]}],"id":[{"id":"https:\/\/ror.org\/05k0wa595","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Orthoflaviviruses comprise genetically diverse mosquito-borne viruses responsible for a broad spectrum of human diseases. Although naturally circulating clinical isolates exhibit biological variability, the extent to which they generate distinct early epithelial innate immune responses remains incompletely understood. Here, we characterized five clinical orthoflavivirus isolates obtained in Oaxaca, Mexico, using human HaCaT keratinocytes as an in vitro model of early infection. Productive infection was assessed by immunofluorescence microscopy, immunostained focus appearance under isolate-optimized assay conditions, and infectious virus production, whereas host responses were evaluated by transcriptional profiling and quantitative whole-slide single-cell immunofluorescence. All isolates established productive infection and exhibited different viral replication profiles. Temporal transcriptional analyses revealed variable expression of antiviral (IFN\u03b2, Mx1, OAS1, PKR, IFITM3, Viperin, and RANTES) and inflammatory (TNF-\u03b1, IL-8, and MCP-1) genes. Quantitative whole-slide analysis provided complementary evidence of variable STAT1 and NF-\u03baB signaling activation across the analyzed isolates. Within this limited panel, viral replication was not consistently aligned with the selected transcriptional and signaling readouts, although the exploratory nature of these comparisons precludes establishing independence between these variables. Together, the virological, transcriptional, and imaging analyses revealed distinct multidimensional functional profiles across the isolate panel. Overall, these findings demonstrate functional heterogeneity among the analyzed clinical orthoflavivirus isolates and highlight integrated functional phenotyping as a useful framework for examining virus\u2013host interactions beyond viral replication alone.<\/jats:p>","DOI":"10.3390\/v18080826","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T16:11:56Z","timestamp":1785168716000},"page":"826","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Functional Characterization of a Panel of Clinical Orthoflavivirus Isolates Reveals Distinct Replication and Innate Immune Response Profiles in Human Keratinocytes"],"prefix":"10.3390","volume":"18","author":[{"given":"Tannya Karen","family":"Castro Jim\u00e9nez","sequence":"first","affiliation":[{"name":"Laboratorio de Biolog\u00eda Molecular e Inmunolog\u00eda de arbovirus, Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Tlalnepantla 54090, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-5475-1960","authenticated-orcid":false,"given":"Edwin Antonio","family":"Lopez Kelly","sequence":"additional","affiliation":[{"name":"Laboratorio de Biolog\u00eda Molecular e Inmunolog\u00eda de arbovirus, Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Tlalnepantla 54090, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2642-3872","authenticated-orcid":false,"given":"Leticia","family":"Cedillo-Barr\u00f3n","sequence":"additional","affiliation":[{"name":"Departamento de Biomedicina Molecular, Centro de Investigaci\u00f3n y de Estudios Avanzados del Instituto Polit\u00e9cnico Nacional, Zacatenco 07360, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3369-8591","authenticated-orcid":false,"given":"Julio","family":"Garc\u00eda-Cordero","sequence":"additional","affiliation":[{"name":"Departamento de Biomedicina Molecular, Centro de Investigaci\u00f3n y de Estudios Avanzados del Instituto Polit\u00e9cnico Nacional, Zacatenco 07360, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4535-5262","authenticated-orcid":false,"given":"Diego Sait","family":"Cruz-Hern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Departamento de Ciencias Biom\u00e9dicas, Universidad de la Sierra Sur, Guillermo Rojas Mijangos s\/n, Esq. AV. Universidad, Col. Ciudad Universitaria, Miahuatl\u00e1n de Porfirio D\u00edaz, Oaxaca 70800, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nallely","family":"Diaz Lima","sequence":"additional","affiliation":[{"name":"OaxacaLab Laboratorio de An\u00e1lisis Cl\u00ednicos, Oaxaca 68000, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Alberto","family":"San Juan Luis","sequence":"additional","affiliation":[{"name":"OaxacaLab Laboratorio de An\u00e1lisis Cl\u00ednicos, Oaxaca 68000, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-9779-1111","authenticated-orcid":false,"given":"Cruz Carlos","family":"Castillo Camacho","sequence":"additional","affiliation":[{"name":"Laboratorio Ecolog\u00eda Microbiana, UBIPRO, Facultad de Estudios Superiores Iztacala, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Tlalnepantla 54090, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2876-9463","authenticated-orcid":false,"given":"Eloy Andr\u00e9s","family":"P\u00e9rez-Y\u00e9pez","sequence":"additional","affiliation":[{"name":"Laboratorio de Se\u00f1alizaci\u00f3n Celular e Inmunolog\u00eda del C\u00e1ncer (SECIC), Instituto Nacional de Cancerolog\u00eda, San Fernando 22, Col. Secci\u00f3n XVI, Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cynthia Daniela","family":"Ibarra-Moreno","sequence":"additional","affiliation":[{"name":"Laboratorio de Se\u00f1alizaci\u00f3n Celular e Inmunolog\u00eda del C\u00e1ncer (SECIC), Instituto Nacional de Cancerolog\u00eda, San Fernando 22, Col. Secci\u00f3n XVI, Tlalpan, Ciudad de M\u00e9xico 14080, Mexico"},{"name":"Escuela de Medicina y Ciencias de la Salud, Tecnol\u00f3gico de Monterrey, Ciudad de M\u00e9xico 14380, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8157-5233","authenticated-orcid":false,"given":"Luis Angel","family":"Flores-Mej\u00eda","sequence":"additional","affiliation":[{"name":"Laboratorio de Falla Medular y Carcinog\u00e9nesis, Departamento de Medicina Gen\u00f3mica y Toxicolog\u00eda Ambiental, Sede Perif\u00e9rica del Instituto de Investigaciones Biom\u00e9dicas, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Coyoac\u00e1n, Ciudad de M\u00e9xico 04530, Mexico"},{"name":"Torre de Investigaci\u00f3n del Instituto Nacional de Pediatr\u00eda, Av. del Im\u00e1n No1, Insurgentes Cuicuilco, Coyoac\u00e1n, Ciudad de M\u00e9xico 04530, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2412-0360","authenticated-orcid":false,"given":"Sergio Roberto","family":"Aguilar-Ru\u00edz","sequence":"additional","affiliation":[{"name":"Departamento de Biomedicina Experimental, Facultad de Medicina y Cirug\u00eda de la Universidad Aut\u00f3noma \u2018Benito Ju\u00e1rez\u2019 de Oaxaca, Oaxaca 68120, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3056-9997","authenticated-orcid":false,"given":"M\u00f3nica G.","family":"Mendoza-Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Avenida de los Barrios 1, Los Reyes Iztacala, Tlalnepantla 54090, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4649-4117","authenticated-orcid":false,"given":"Luis I.","family":"Terrazas","sequence":"additional","affiliation":[{"name":"Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Avenida de los Barrios 1, Los Reyes Iztacala, Tlalnepantla 54090, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7368-6432","authenticated-orcid":false,"given":"Jos\u00e9","family":"Bustos-Arriaga","sequence":"additional","affiliation":[{"name":"Laboratorio de Biolog\u00eda Molecular e Inmunolog\u00eda de arbovirus, Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Tlalnepantla 54090, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1007\/s00705-023-05835-1","article-title":"Renaming of the Genus Flavivirus to Orthoflavivirus and Extension of Binomial Species Names within the Family Flaviviridae","volume":"168","author":"Postler","year":"2023","journal-title":"Arch. 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