{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:44:23Z","timestamp":1791333863581,"version":"4.3.1"},"reference-count":77,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,7,16]],"date-time":"2024-07-16T00:00:00Z","timestamp":1721088000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Variant pseudorabies virus (PRV) is a newly emerged zoonotic pathogen that can cause human blindness. PRV can take advantage of its large genome and multiple non-essential genes to construct recombinant attenuated vaccines carrying foreign genes. However, a major problem is that the foreign genes in recombinant PRV are only integrated into the genome for independent expression, rather than assembled on the surface of virion.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We reported a recombinant PRV with deleted gE\/TK genes and an inserted porcine circovirus virus 2 (PCV2) <jats:italic>Cap<\/jats:italic> gene into the extracellular domain of the PRV gE gene using the Cre-loxP recombinant system combined with the CRISPR-Cas9 gene editing system. This recombinant PRV (PRV-Cap), with the envelope-embedded Cap protein, exhibits a similar replication ability to its parental virus.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>An immunogenicity assay revealed that PRV-Cap immunized mice have 100% resistance to lethal PRV and PCV2 attacks. Neutralization antibody and ELISPOT detections indicated that PRV-Cap can enhance neutralizing antibodies to PRV and produce IFN-\u03b3 secreting T cells specific for both PRV and PCV2. Immunological mechanistic investigation revealed that initial immunization with PRV-Cap stimulates significantly early activation and expansion of CD69<jats:sup>+<\/jats:sup> T cells, promoting the activation of CD4 Tfh cell dependent germinal B cells and producing effectively specific effector memory T and B cells. Booster immunization with PRV-Cap recalled the activation of PRV-specific IFN-\u03b3<jats:sup>+<\/jats:sup>IL-2<jats:sup>+<\/jats:sup>CD4<jats:sup>+<\/jats:sup> T cells and IFN-\u03b3<jats:sup>+<\/jats:sup>TNF-\u03b1<jats:sup>+<\/jats:sup>CD8<jats:sup>+<\/jats:sup> T cells, as well as PCV2-specific IFN-\u03b3<jats:sup>+<\/jats:sup>TNF-\u03b1<jats:sup>+<\/jats:sup>CD8<jats:sup>+<\/jats:sup> T cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Collectively, our data suggested an immunological mechanism in that the recombinant PRV with envelope-assembled PCV2 Cap protein can serve as an excellent vaccine candidate for combined immunity against PRV and PCV2, and provided a cost-effective method for the production of PRV- PCV2 vaccine.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2024.1438371","type":"journal-article","created":{"date-parts":[[2024,7,16]],"date-time":"2024-07-16T04:49:37Z","timestamp":1721105377000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Immunological characteristics of a recombinant alphaherpesvirus with an envelope-embedded Cap protein of 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