{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T02:50:48Z","timestamp":1760151048289,"version":"build-2065373602"},"reference-count":50,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,16]],"date-time":"2022-02-16T00:00:00Z","timestamp":1644969600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003593","name":"National Council for Scientific and Technological Development","doi-asserted-by":"publisher","award":["303671\/2019-0","314366\/2020-2","421867\/2018-3","406868\/2016-6"],"award-info":[{"award-number":["303671\/2019-0","314366\/2020-2","421867\/2018-3","406868\/2016-6"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfeicoamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["001"],"award-info":[{"award-number":["001"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>The significant number of people with latent and active tuberculosis infection requires further efforts to develop new vaccines or improve the Bacillus Calmette-Gu\u00e9rin (BCG), which is the only approved vaccine against this disease. In this study, we developed a recombinant fusion protein (PEPf) containing high-density immunodominant epitope sequences from Rv0125, Rv2467, and Rv2672 Mycobacterium tuberculosis (Mtb) proteases that proved immunogenic and used it to develop a recombinant BCG vaccine expressing the fusion protein. After challenging using Mtb, a specific immune response was recalled, resulting in a reduced lung bacterial load with similar protective capabilities to BCG. Thus BCG PEPf failed to increase the protection conferred by BCG. The PEPf was combined with Advax4 adjuvant and tested as a subunit vaccine using a prime-boost strategy. PEPf + Advax4 significantly improved protection after Mtb challenge, with a reduction in bacterial load in the lungs. Our results confirm that Mtb proteases can be used to develop vaccines against tuberculosis and that the use of the recombinant PEPf subunit protein following a prime-boost regimen is a promising strategy to improve BCG immunity.<\/jats:p>","DOI":"10.3390\/vaccines10020306","type":"journal-article","created":{"date-parts":[[2022,2,16]],"date-time":"2022-02-16T20:28:59Z","timestamp":1645043339000},"page":"306","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Protease-Based Subunit Vaccine in Mice Boosts BCG Protection against Mycobacterium tuberculosis"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4200-6617","authenticated-orcid":false,"given":"Ana Paula","family":"Junqueira-Kipnis","sequence":"first","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carine","family":"de Castro Souza","sequence":"additional","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Carolina","family":"de Oliveira Carvalho","sequence":"additional","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4470-8444","authenticated-orcid":false,"given":"Fabio Muniz","family":"de Oliveira","sequence":"additional","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3635-7787","authenticated-orcid":false,"given":"Vinnycius Pereira","family":"Almeida","sequence":"additional","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alisson Rodrigues","family":"de Paula","sequence":"additional","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4733-007X","authenticated-orcid":false,"given":"Mara Rubia","family":"Celes","sequence":"additional","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5950-106X","authenticated-orcid":false,"given":"Andr\u00e9","family":"Kipnis","sequence":"additional","affiliation":[{"name":"Department of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goi\u00e1s, Goi\u00e2nia 74605-050, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1093\/trstmh\/trw016","article-title":"TB Vaccine Development and the End TB Strategy: Importance and Current Status","volume":"110","author":"Fletcher","year":"2016","journal-title":"Trans. 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