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The incorporation of mutations that stabilize analogous proteins from other viruses in their prefusion conformation (e.g., HIV Env, SARS-CoV-2 S, or RSV F glycoproteins) has improved their capability to induce neutralizing protective immune responses. Therefore, we have stabilized the FeLV Env protein following a strategy based on the incorporation of a disulfide bond and an Ile\/Pro mutation (SOSIP) previously used to generate soluble HIV Env trimers. We have characterized this SOSIP-FeLV Env in its soluble form and as a transmembrane protein present at high density on the surface of FeLV Gag-based VLPs. Furthermore, we have tested its immunogenicity in DNA-immunization assays in C57BL\/6 mice. Low anti-FeLV Env responses were detected in SOSIP-FeLV soluble protein-immunized animals; however, unexpectedly no responses were detected in the animals immunized with SOSIP-FeLV Gag-based VLPs. In contrast, high humoral response against FeLV Gag was observed in the animals immunized with control Gag VLPs lacking SOSIP-FeLV Env, while this response was significantly impaired when the VLPs incorporated SOSIP-FeLV Env. Our data suggest that FeLV Env can be stabilized as a soluble protein and can be expressed in high-density VLPs. However, when formulated as a DNA vaccine, SOSIP-FeLV Env remains poorly immunogenic, a limitation that must be overcome to develop an effective FeLV vaccine.<\/jats:p>","DOI":"10.3390\/v16060987","type":"journal-article","created":{"date-parts":[[2024,6,21]],"date-time":"2024-06-21T11:10:28Z","timestamp":1718968228000},"page":"987","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Production and Immunogenicity of FeLV Gag-Based VLPs Exposing a Stabilized FeLV Envelope Glycoprotein"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6919-2026","authenticated-orcid":false,"given":"Raquel","family":"Ortiz","sequence":"first","affiliation":[{"name":"IrsiCaixa, 08916 Badalona, Spain"},{"name":"Doctorate School, Microbiology Department, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, 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Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bonaventura","family":"Clotet","sequence":"additional","affiliation":[{"name":"IrsiCaixa, 08916 Badalona, Spain"},{"name":"Centre for Health and Social Care Research (CESS), Faculty of Medicine, University of Vic-Central University of Catalonia (UVic-UCC), 08500 Vic, Spain"},{"name":"Infectious Diseases Department, Germans Trias I Pujol Hospital, 08916 Badalona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jorge","family":"Carrillo","sequence":"additional","affiliation":[{"name":"IrsiCaixa, 08916 Badalona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5701-7650","authenticated-orcid":false,"given":"Carmen","family":"Aguilar-Gurrieri","sequence":"additional","affiliation":[{"name":"IrsiCaixa, 08916 Badalona, 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