{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,26]],"date-time":"2026-10-26T21:36:11Z","timestamp":1793050571637,"version":"4.5.0"},"reference-count":12,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T00:00:00Z","timestamp":1787097600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T00:00:00Z","timestamp":1787097600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100004763","name":"Natural Science Foundation of Inner Mongolia Autonomous Region","doi-asserted-by":"publisher","award":["2023ZD15"],"award-info":[{"award-number":["2023ZD15"]}],"id":[{"id":"10.13039\/501100004763","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Braz J Microbiol"],"published-print":{"date-parts":[[2026,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Brucellosis is one of the most severe Class B infectious diseases prevalent in the agricultural and pastoral areas of northern China. Current attenuated live vaccines (e.g., M5, S19) have defects such as residual virulence, causing abortion in pregnant animals, and the inability to differentiate between natural infection and vaccination (DIVA).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>In this study, published public datasets of C57BL\/6J mice chronic brucellosis infection were adopted for single-cell transcriptome sequencing (10x Genomics) analysis and re-analysed to map the heterogeneity of splenic immune cells. Whole-genome scanning and pangenomic analysis were performed on four strains with different virulence levels (M16, 544\u00a0A, M5, 104\u00a0M) using second-generation sequencing public data. Membrane\/secreted proteins unique and conserved in virulent strains were screened as candidate antigens, prepared via prokaryotic expression systems, and their humoral and cellular immune levels were detected by indirect ELISA and flow cytometry.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Published public chronic infection dataset showed stable chronic infection characteristics (bacterial load Log10 CFU\u2009&gt;\u20094.5). Single-cell sequencing yielded 45,231 cells, annotated into 12 immune cell subpopulations, revealing significant expansion of Effector CD4\u2009+\u2009T cells (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.01) and high expression of the\n                      <jats:italic>Ifng<\/jats:italic>\n                      gene. Pangenomic analysis identified three candidate antigens (BMEI0021, BMEI1943, BMEI0367) that are 100% conserved in virulent strains but absent in the vaccine strain M5. Immunogenicity assays showed that BMEI1943 induced high levels of IgG2a subtype antibodies (titer Log2 13.45\u2009\u00b1\u20090.52) and IFN-\u03b3\u2009+\u2009CD4\u2009+\u2009T cell responses (frequency 15.80% \u00b1 2.10%).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Through multi-omics integration analysis, this study successfully identified a novel candidate antigen, BMEI1943, with strong Th1-type immunogenicity, providing an experimental basis for the development of safe and efficient subunit vaccines against brucellosis.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s42770-026-02049-w","type":"journal-article","created":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T12:31:45Z","timestamp":1787142705000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated multi-omics strategy for screening novel protective antigens against Brucella melitensis and analysis of their immunogenicity"],"prefix":"10.1007","volume":"57","author":[{"given":"Zhiheng","family":"Dong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sha","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiarong","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-8716-6068","authenticated-orcid":false,"given":"Lidao","family":"Bao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,19]]},"reference":[{"issue":"1","key":"2049_CR1","doi-asserted-by":"publisher","DOI":"10.1186\/s12879-024-10165-x","volume":"24","author":"S Liu","year":"2024","unstructured":"Liu S, Soontornchai S, Bovornkitti S, Wang X (2024) Epidemiological characteristics and spatio-temporal clusters of human brucellosis in Inner Mongolia, 2010\u20132021. 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Transbound Emerg Dis 69(5):e587\u2013e601","journal-title":"Transbound Emerg Dis"},{"issue":"8","key":"2049_CR4","doi-asserted-by":"publisher","first-page":"ftaa002","DOI":"10.1093\/femspd\/ftaa002","volume":"77","author":"S Gupta","year":"2019","unstructured":"Gupta S, Singh D, Gupta M, Bhatnagar R (2019) A combined subunit vaccine comprising BP26, Omp25 and L7\/L12 against brucellosis. Pathogens Disease 77(8):ftaa002","journal-title":"Pathogens Disease"},{"issue":"1","key":"2049_CR5","doi-asserted-by":"publisher","first-page":"5037","DOI":"10.1038\/s41467-018-07492-4","volume":"9","author":"G Xin","year":"2018","unstructured":"Xin G, Zander R, Schauder DM et al (2018) Single-cell RNA sequencing unveils an IL-10-producing helper subset that sustains humoral immunity during persistent infection. 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May 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 August 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 August 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 October 2026","order":5,"name":"change_date","label":"Change Date","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Update","order":6,"name":"change_type","label":"Change Type","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The original online version of this article was revised:","order":7,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"In the Abstract, Introduction, Methods, and Results sections of this article, references to the chronic infection model involving live *Brucella melitensis* and the associated experimental data should be removed.","order":8,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"In this article, the description of single-cell transcriptome sequencing performed on samples derived from infected animals should be removed, including the corresponding methodological and results descriptions.","order":9,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"In this article, references to P3\/BSL-3 containment facilities and to the Inner Mongolia Center for Disease Control and Prevention should be removed.","order":10,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"In this article, the related biosafety and research-conduct statement, ethical statement, conflict-of-interest statement, and author-contribution statement should be removed or amended accordingly.","order":11,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"In this article, the keywords \u201cABSL-3\u201d and \u201cSingle-cell transcriptome\u201d should be removed.","order":12,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"In this article, the title should be amended to reflect the revised scope of the study following removal of the single-cell transcriptome component.","order":13,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The remaining study components, including comparative genomics and pangenomic analyses, recombinant protein expression and purification, immunization with recombinant proteins and heat-inactivated M5 lysate, ELISA measurements, and flow cytometric detection of intracellular IFN-\u03b3, remain unchanged.","order":14,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The original article has been corrected.","order":15,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 October 2026","order":16,"name":"change_date","label":"Change Date","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Correction","order":17,"name":"change_type","label":"Change Type","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"A Correction to this paper has been published:","order":18,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"https:\/\/doi.org\/10.1007\/s42770-026-02099-0","URL":"https:\/\/doi.org\/10.1007\/s42770-026-02099-0","order":19,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests regarding the publication of this manuscript.","order":1,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"244"}}