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Bacillus Calmette\u2013Gu\u00e9rin (BCG), the existing tuberculosis vaccine, is known to induce heterologous effects over other infections due to trained immunity and has been proposed to be a potential strategy against SARS-CoV-2 infection. In this report, we constructed a recombinant BCG (rBCG) expressing domains of the SARS-CoV-2 nucleocapsid and spike proteins (termed rBCG-ChD6), recognized as major candidates for vaccine development. We investigated whether rBCG-ChD6 immunization followed by a boost with the recombinant nucleocapsid and spike chimera (rChimera), together with alum, provided protection against SARS-CoV-2 infection in K18-hACE2 mice. A single dose of rBCG-ChD6 boosted with rChimera associated with alum elicited the highest anti-Chimera total IgG and IgG2c Ab titers with neutralizing activity against SARS-CoV-2 Wuhan strain when compared with control groups. Importantly, following SARS-CoV-2 challenge, this vaccination regimen induced IFN-\u03b3 and IL-6 production in spleen cells and reduced viral load in the lungs. In addition, no viable virus was detected in mice immunized with rBCG-ChD6 boosted with rChimera, which was associated with decreased lung pathology when compared with BCG WT-rChimera\/alum or rChimera\/alum control groups. Overall, our study demonstrates the potential of a prime-boost immunization system based on an rBCG expressing a chimeric protein derived from SARS-CoV-2 to protect mice against viral challenge.<\/jats:p>","DOI":"10.4049\/jimmunol.2200731","type":"journal-article","created":{"date-parts":[[2023,4,26]],"date-time":"2023-04-26T09:57:57Z","timestamp":1682503077000},"page":"1925-1937","source":"Crossref","is-referenced-by-count":20,"title":["Recombinant Bacillus Calmette\u2013Gu\u00e9rin Expressing SARS-CoV-2 Chimeric Protein Protects K18-hACE2 Mice against Viral Challenge"],"prefix":"10.1093","volume":"210","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8956-7219","authenticated-orcid":false,"given":"F\u00e1bio","family":"Mambelli","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6885-3438","authenticated-orcid":false,"given":"F\u00e1bio V","family":"Marinho","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-0100-0027","authenticated-orcid":false,"given":"Juvana M","family":"Andrade","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1045-2249","authenticated-orcid":false,"given":"Ana C V S C","family":"de Araujo","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]},{"name":"Department of Genetics, Ecology and Evolution, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo P F","family":"Abuna","sequence":"additional","affiliation":[{"name":"Platform of Bi-Institutional Research in Translational Medicine, Oswaldo Cruz Foundation-Fiocruz , Ribeir\u00e3o Preto, S\u00e3o Paulo,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7300-537X","authenticated-orcid":false,"given":"Victor M R","family":"Fabri","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1854-8907","authenticated-orcid":false,"given":"Bruno P O","family":"Santos","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00e3o S","family":"da Silva","sequence":"additional","affiliation":[{"name":"Platform of Bi-Institutional Research in Translational Medicine, Oswaldo Cruz Foundation-Fiocruz , Ribeir\u00e3o Preto, S\u00e3o Paulo,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4912-0233","authenticated-orcid":false,"given":"Mariana T Q","family":"de Magalh\u00e3es","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2685-8852","authenticated-orcid":false,"given":"E Jane","family":"Homan","sequence":"additional","affiliation":[{"name":"ioGenetics LLC , Madison, WI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciana C C","family":"Leite","sequence":"additional","affiliation":[{"name":"Vaccine Development Laboratory, Butantan Institute , S\u00e3o Paulo, SP,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Greicy B M","family":"Dias","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florian\u00f3polis , Santa Catarina,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicoli","family":"Heck","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florian\u00f3polis , Santa Catarina,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5655-0931","authenticated-orcid":false,"given":"Daniel A G B","family":"Mendes","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florian\u00f3polis , Santa Catarina,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel S","family":"Mansur","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florian\u00f3polis , Santa Catarina,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9","family":"B\u00e1fica","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florian\u00f3polis , Santa Catarina,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4062-5577","authenticated-orcid":false,"given":"Sergio C","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais , Belo Horizonte, Minas Gerais,","place":["Brazil"]},{"name":"Department of Immunology, Institute of Biomedical Science, University of S\u00e3o Paulo , S\u00e3o Paulo,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,4,26]]},"reference":[{"key":"2026041420283654100_r1","article-title":"Johns Hopkins University & Medicine Database","author":"Coronavirus Resource Center, Center for Systems Science and Engineering"},{"key":"2026041420283654100_r2","article-title":"WHO coronavirus (COVID-19) dashboard","author":"World Health Organization (WHO)"},{"key":"2026041420283654100_r3","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1056\/NEJMoa2001017","article-title":"A novel coronavirus from patients with pneumonia in China, 2019","volume":"382","author":"Zhu","year":"2020","journal-title":"N. 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Pharmacol."},{"key":"2026041420283654100_r9","doi-asserted-by":"crossref","first-page":"e200342","DOI":"10.1590\/0074-02760200342","article-title":"SARS-CoV-2 isolation from the first reported patients in Brazil and establishment of a coordinated task network","volume":"115","author":"Araujo","year":"2020","journal-title":"Mem. Inst. Oswaldo Cruz"},{"key":"2026041420283654100_r10","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.jhep.2020.09.031","article-title":"COVID-19: discovery, diagnostics and drug development","volume":"74","author":"Asselah","year":"2021","journal-title":"J. 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[Published erratum appears in 2020 Cell 183: 1735.]","volume":"181","author":"Walls","year":"2020","journal-title":"Cell"},{"key":"2026041420283654100_r16","doi-asserted-by":"crossref","first-page":"206","DOI":"10.3390\/cells10020206","article-title":"COVID-19: characteristics and therapeutics","volume":"10","author":"Chilamakuri","year":"2021","journal-title":"Cells"},{"key":"2026041420283654100_r17","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1007\/s10096-020-04138-6","article-title":"COVID-19: angiotensin-converting enzyme 2 (ACE2) expression and tissue susceptibility to SARS-CoV-2 infection","volume":"40","author":"Beyerstedt","year":"2021","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"2026041420283654100_r18","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1002\/jmv.26232","article-title":"The cytokine storm and COVID-19","volume":"93","author":"Hu","year":"2021","journal-title":"J. Med. 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