{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T23:03:36Z","timestamp":1789945416410,"version":"4.0.1"},"reference-count":22,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T00:00:00Z","timestamp":1782345600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"vor","delay-in-days":88,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Injectable inactivated whole-cell cholera vaccines provide limited protection due to poor mucosal immunity, leading to a shift toward oral cholera vaccines (OCVs). Chimeric recombinant vaccines can deliver multiple antigens simultaneously, enhancing immune specificity and simplifying production. This study aimed to develop a PLGA-encapsulated OCV using a chimeric protein for enhanced mucosal delivery and immune response. A synthetic gene encoding r63 was cloned into pET28a and expressed in\n                    <jats:italic>E. coli<\/jats:italic>\n                    BL21(DE3). The ~\u200963\u00a0kDa His-tagged chimeric protein (r63) was purified via Ni-NTA affinity chromatography and entrapped in PLGA nanoparticles using a double emulsion-solvent evaporation technique. Particle size, zeta potential, SEM morphology, protein release kinetics, and encapsulation efficiency were evaluated. BALB\/c mice were orally immunized with r63-PLGA, and IgG\/IgA responses were measured by ELISA. Functional assays, including GM1-ELISA, Y-1 cell rounding, cAMP inhibition and rabbit ileal loop were performed to assess cholera toxin neutralization. In vitro adherence inhibition of\n                    <jats:italic>V. cholera<\/jats:italic>\n                    was investigated. The r63 was successfully expressed in\n                    <jats:italic>E. coli<\/jats:italic>\n                    and confirmed by SDS-PAGE and Western blotting. PLGA- r63 nanoparticles showed high encapsulation efficiency (90.16\u2009\u00b1\u20093.4%), a size of 221\u2009\u00b1\u200910\u00a0nm, and a zeta potential of \u2212\u200911.8\u2009\u00b1\u20092 mV. Release studies indicated sustained protein release fitting a zero-order kinetic model. Oral immunization elicited robust serum IgG and fecal IgA responses against all antigens. Immunized sera prevented CTB binding to GM1 receptor, neutralized the enterotoxicity of cholera toxin in the Y1 cell line, and inhibited cAMP production. The immunized serum also suppressed cholera toxin activity in the ileal loop test by up to 79% and inhibited 66% of the bacterial adhesion to Caco2 cells. The orally delivered PLGA-r63 candidate vaccine effectively induced systemic and mucosal immunity against adherence and toxicity\n                    <jats:italic>Vibrio cholerae<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-52925-6","type":"journal-article","created":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T09:15:01Z","timestamp":1782378901000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["PLGA-entrapped recombinant r63 chimeric protein induces mucosal immunity against adherence and toxicity of Vibrio cholerae"],"prefix":"10.1038","volume":"16","author":[{"given":"Hossein","family":"Samiei-Abianeh","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shahram","family":"Nazarian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad","family":"Behrozie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emad","family":"Kordbacheh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,25]]},"reference":[{"key":"52925_CR1","doi-asserted-by":"publisher","unstructured":"Montero, D. A. et al. Vibrio cholerae, classification, pathogenesis, immune response, and trends in vaccine development. Front. Med. 10. https:\/\/doi.org\/10.3389\/fmed.2023.1155751 (2023).","DOI":"10.3389\/fmed.2023.1155751"},{"key":"52925_CR2","unstructured":"(WHO). Epidemic and Pandemic-Prone Diseases. https:\/\/www.emro.who.int\/fr\/pandemic-epidemic-diseases\/cholera\/acute-watery-diarrhoeacholera-updates-27-january-2024.html (2024)."},{"key":"52925_CR3","unstructured":"(EDCD), E. C. f. D. P. a. C. Cholera Worldwide Overview. https:\/\/www.ecdc.europa.eu\/en\/all-topics-z\/cholera\/surveillance-and-disease-data\/cholera-monthly (2025)."},{"key":"52925_CR4","doi-asserted-by":"publisher","first-page":"974","DOI":"10.3390\/toxins7030974","volume":"7","author":"KJ Baldauf","year":"2015","unstructured":"Baldauf, K. J., Royal, J. M., Hamorsky, K. T. & Matoba, N. Cholera toxin B: One subunit with many pharmaceutical applications. Toxins 7, 974\u2013996. https:\/\/doi.org\/10.3390\/toxins7030974 (2015).","journal-title":"Toxins"},{"key":"52925_CR5","first-page":"195","volume":"133","author":"B Das","year":"2011","unstructured":"Das, B., Bischerour, J. & Barre, F. X. Molecular mechanism of acquisition of the cholera toxin genes. Indian J. Med. Res. 133, 195\u2013200 (2011).","journal-title":"Indian J. Med. Res."},{"key":"52925_CR6","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1007\/s00294-015-0532-8","volume":"62","author":"SA Jung","year":"2016","unstructured":"Jung, S. A., Hawver, L. A. & Ng, W. L. Parallel quorum sensing signaling pathways in Vibrio cholerae. Curr. Genet. 62, 255\u2013260. https:\/\/doi.org\/10.1007\/s00294-015-0532-8 (2016).","journal-title":"Curr. Genet."},{"key":"52925_CR7","doi-asserted-by":"publisher","first-page":"322","DOI":"10.1016\/j.vaccine.2017.09.004","volume":"36","author":"R Adriani","year":"2018","unstructured":"Adriani, R., Mousavi Gargari, S. L., Nazarian, S., Sarvary, S. & Noroozi, N. Immunogenicity of Vibrio cholerae outer membrane vesicles secreted at various environmental conditions. Vaccine 36, 322\u2013330. https:\/\/doi.org\/10.1016\/j.vaccine.2017.09.004 (2018).","journal-title":"Vaccine"},{"key":"52925_CR8","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1016\/j.jconrel.2022.01.050","volume":"343","author":"X He","year":"2022","unstructured":"He, X. et al. A potential delivery system based on cholera toxin: A macromolecule carrier with multiple activities. J. Control. Release 343, 551\u2013563. https:\/\/doi.org\/10.1016\/j.jconrel.2022.01.050 (2022).","journal-title":"J. Control. Release"},{"key":"52925_CR9","doi-asserted-by":"publisher","first-page":"1274","DOI":"10.4161\/hv.28371","volume":"10","author":"J Hou","year":"2014","unstructured":"Hou, J. et al. Cholera toxin B subunit acts as a potent systemic adjuvant for HIV-1 DNA vaccination intramuscularly in mice. Hum. Vac. 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