{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,31]],"date-time":"2026-01-31T10:08:46Z","timestamp":1769854126025,"version":"3.49.0"},"reference-count":62,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T00:00:00Z","timestamp":1744675200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T00:00:00Z","timestamp":1744675200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"Chemical Industries Education & Training Authority","award":["Learning-Contract-1171"],"award-info":[{"award-number":["Learning-Contract-1171"]}]},{"name":"Faculty of National and Agricultural Sciences, University of Pretoria"},{"name":"Onderstepoort Biological Products"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2025,4,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>Control of bluetongue disease is predominantly through vaccination with licensed inactivated or live-attenuated vaccines (LAVs). Manufacturing of LAVs in endemic countries requires formulation with a high number of serotypes for effective protection. Herein, we evaluated a plasmid DNA-based reverse genetics platform for manufacturing a multivalent vaccine. The synthetic vaccine was characterized by a common BTV1 backbone, with exchange of outer capsid proteins. Recombinant South African vaccine serotypes 1, 5, and 14 were rescued by exchanging the VP2 protein on the backbone. BTV6 rescue was achieved following the exchange of VP2 and VP5 proteins. The particle sizes were comparable to commercial vaccines of respective serotypes. BTV1, BTV5, and BTV6 had distinct growth profiles compared to commercial vaccines, while BTV14 was indistinguishable. Stability and shelf-life determination under various storage conditions showed that commercial vaccines were more stable. Formulated antigens were evaluated for vaccine safety and immunogenicity in sheep. Serotyped BTV1 monovalent vaccine was safe, as no clinical signs were observed. Neutralizing antibodies (nAbs) were induced on day 14 and peaked at 32 on day 28. The multivalent synthetic vaccine containing four serotypes elicited BTV6 nAbs from day 21 with a titer of 52, which decreased to 33 by day 42. BTV1 elicited a weak immune response with a titer of 1 on day 42. No nAbs were detected against BTV5 and BTV14. This is a first report comparing reverse genetics-derived antigens with commercial vaccines. Data generated on production yields, stability, and immunogenicity demonstrated that some serotypes can be implemented as novel synthetic vaccines using this platform.<\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>Vaccination is the most effective control strategy for viral diseases that affect livestock. To date, only live-attenuated and inactivated vaccines have been licensed for control of bluetongue (BT). This study demonstrated the use of reverse genetics as a possible platform for BTV vaccine production. Data generated in the study contribute toward the advancement of an alternative manufacturing platform for licensing of BT vaccines. Information on production yields and stability of synthetic vaccines in comparison to the conventional products demonstrated that optimization is required for some serotypes to fully translate the reverse genetics platform for manufacturing the BTV vaccine. The study highlighted the safety and immunogenicity of vaccines manufactured using the plasmid DNA-based reverse-genetics platform.<\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/jvi.00139-25","type":"journal-article","created":{"date-parts":[[2025,3,6]],"date-time":"2025-03-06T14:00:13Z","timestamp":1741269613000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Plasmid DNA-based reverse genetics as a platform for manufacturing of bluetongue vaccine"],"prefix":"10.1128","volume":"99","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0273-8933","authenticated-orcid":true,"given":"Tendai A. 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