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Accordingly, the development of effective vaccines against\n                      <jats:italic>T. gondii<\/jats:italic>\n                      has become an urgent priority. In this study, we constructed a multi-epitope chimeric antigen T-SGR targeting three key protective antigens of\n                      <jats:italic>T. gondii<\/jats:italic>\n                      (SAG1, GRA7, and ROP16), and developed both a messenger RNA (mRNA) lipid nanoparticle (LNP) vaccine and a recombinant protein vaccine based on T-SGR. The immunogenicity and protective effects were further evaluated in C57BL\/6 mice.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      The T-SGR mRNA\u2013LNP vaccine was prepared via in vitro transcription followed by LNP encapsulation, while the T-SGR protein vaccine was obtained via prokaryotic expression and purification. Mice were administered a two-dose immunization regimen. Serum levels of specific IgG, IgG1, and IgG2a antibodies and cytokine levels were measured by enzyme-linked immunosorbent assay (ELISA). T lymphocyte subsets and lymphocyte proliferation were assessed by flow cytometry and Cell Counting Kit-8 (CCK-8) assay. Protective efficacy was evaluated by monitoring survival rates after challenge with highly virulent\n                      <jats:italic>T. gondii<\/jats:italic>\n                      RH strain tachyzoites and moderately virulent ME49 strain tachyzoites.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Both T-SGR mRNA and protein vaccines induced robust humoral and cellular immune responses in mice. Notably, the IgG antibody titer induced by the mRNA\u2013LNP vaccine was significantly higher than that of the protein vaccine (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.05). Both vaccines drove a Th1-biased immune response, as evidenced by markedly higher IgG2a levels relative to IgG1. Compared with the phosphate-buffered saline (PBS) control group, both vaccine groups significantly promoted splenocyte proliferation (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.05). The mRNA vaccine induced significantly higher secretion of IFN-\u03b3, IL-10, IL-12, and IL-2 than the protein vaccine. Both vaccines conferred significant protection against\n                      <jats:italic>T. gondii<\/jats:italic>\n                      infection and prolonged mouse survival. Strikingly, the T-SGR mRNA\u2013LNP vaccine provided 100% protection against the\n                      <jats:italic>T. gondii<\/jats:italic>\n                      ME49 strain, outperforming the recombinant protein vaccine.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      We successfully developed a multi-epitope T-SGR mRNA\u2013LNP vaccine and a recombinant protein vaccine against\n                      <jats:italic>T. gondii<\/jats:italic>\n                      . The T-SGR mRNA\u2013LNP vaccine elicited stronger humoral and cellular immune responses and conferred superior protective efficacy, representing a promising candidate vaccine against toxoplasmosis.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-026-07447-5","type":"journal-article","created":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T20:03:41Z","timestamp":1780689821000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Multi-epitope mRNA vaccine and protein vaccine protect mice against Toxoplasma gondii"],"prefix":"10.1186","volume":"19","author":[{"given":"Yulin","family":"Cui","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongnan","family":"Qu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunxue","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyue","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huaiyu","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hua","family":"Cong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,5]]},"reference":[{"key":"7447_CR1","doi-asserted-by":"publisher","DOI":"10.1128\/cmr.05013-11","author":"F Robert-Gangneux","year":"2012","unstructured":"Robert-Gangneux F, Dard\u00e9 M. 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The maintenance and care of experimental animals were carried out in strict accordance with the regulations of the Animal Ethics Procedures and Guidelines of the People\u2019s Republic of China. All efforts have been made to alleviate suffering of animals used in the study.","order":1,"name":"Ethics","label":"Ethical approval and consent to participate","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","label":"Consent for publication","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","label":"Competing interest","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"329"}}