{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T05:36:04Z","timestamp":1767245764029,"version":"3.48.0"},"reference-count":42,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T00:00:00Z","timestamp":1767052800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100013409","name":"Sistema General de Regal\u00edas, Pontificia Universidad Javeriana","doi-asserted-by":"publisher","award":["2020000100360"],"award-info":[{"award-number":["2020000100360"]}],"id":[{"id":"10.13039\/501100013409","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020905","name":"Universidad Surcolombiana","doi-asserted-by":"crossref","award":["3660"],"award-info":[{"award-number":["3660"]}],"id":[{"id":"10.13039\/501100020905","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Severe dengue virus (DENV) infections are associated with circulating non-neutralizing antibodies generated during heterotypic infections. Although antibodies are key mediators of both protection and pathogenesis, the specific dynamics of B cells (Bc) and their antibody responses remain insufficiently characterized due to limited methods of identifying DENV-specific Bc (DENV-Bc) and the absence of animal models resembling the human disease. Here, we developed a spectral flow cytometry assay employing biotinylated virus-like particles (VLPs) to detect DENV-Bc in C57BL\/6 mice and children hospitalized with dengue. DENV-1 and DENV-2 VLPs were biotinylated, and the efficiency of biotin incorporation was assessed with an HABA-avidin assay and ELISA. Serotype specificity and optimal binding conditions were confirmed using hybridomas 4G2 (pan-flavivirus) and 3H5-1 (DENV-2 specific). Fluorescent agglutimers were subsequently generated by coupling biotinylated VLPs to streptavidin\u2013fluorochrome complexes. Splenocytes from intraperitoneally DENV-infected mice and peripheral blood mononuclear cells (PBMCs) from naturally infected pediatric patients were stained with these VLPs and Bc-lineage markers. Biotinylated VLPs bound specifically to hybridomas, and this binding was competitively inhibited by unlabeled VLPs. After secondary DENV challenge, VLPs identified DENV-specific class-switched plasmablasts in mice. Circulating DENV-specific plasmablasts were also detected in children, with agglutimers enabling the discrimination of serotype-specific and cross-reactive responses in primary and secondary infections. This VLP-based approach represents a scalable platform to investigate the protective and pathogenic roles of DENV-Bc in infection and vaccination.<\/jats:p>","DOI":"10.3390\/v18010058","type":"journal-article","created":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T16:34:23Z","timestamp":1767198863000},"page":"58","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Virus-like Particles and Spectral Flow Cytometry for Identification of Dengue Virus-Specific B Cells in Mice and Humans"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0947-5112","authenticated-orcid":false,"given":"Katherine","family":"Segura","sequence":"first","affiliation":[{"name":"Instituto de Gen\u00e9tica Humana, Facultad de Medicina, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"},{"name":"Divisi\u00f3n de Inmunolog\u00eda, Programa de Medicina, Facultad de Ciencias de la Salud, Universidad Surcolombiana, Neiva 410010, Huila, Colombia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabiola","family":"Martel","sequence":"additional","affiliation":[{"name":"Instituto de Gen\u00e9tica Humana, Facultad de Medicina, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manuel A.","family":"Franco","sequence":"additional","affiliation":[{"name":"Instituto de Gen\u00e9tica Humana, Facultad de Medicina, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0119-8306","authenticated-orcid":false,"given":"Federico","family":"Perdomo-Celis","sequence":"additional","affiliation":[{"name":"Instituto de Gen\u00e9tica Humana, Facultad de Medicina, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0129-5210","authenticated-orcid":false,"given":"Carlos F.","family":"Narv\u00e1ez","sequence":"additional","affiliation":[{"name":"Divisi\u00f3n de Inmunolog\u00eda, Programa de Medicina, Facultad de Ciencias de la Salud, Universidad Surcolombiana, Neiva 410010, Huila, Colombia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1016\/S0140-6736(23)02576-X","article-title":"Dengue","volume":"403","author":"Adams","year":"2024","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"102585","DOI":"10.1016\/j.jiph.2024.102585","article-title":"The Unprecedented Surge of Dengue in the Americas: Strategies for Effective Response","volume":"17","author":"Pareek","year":"2024","journal-title":"J. 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