{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T10:40:00Z","timestamp":1772793600077,"version":"3.50.1"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T00:00:00Z","timestamp":1704931200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The attenuation of BCG has led to the loss of not only immunogenic proteins but also lipid antigens.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Thus, we compared the macrophage and T-cell responses to nonpolar lipid extracts harvested from BCG and <jats:italic>Mycobacterium tuberculosis<\/jats:italic> (<jats:italic>Mtb<\/jats:italic>) to better understand the role of BCG lipids in the already known diminished responses of the vaccine strain.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Relative to Mtb, nonpolar lipid extract from BCG presented a reduced capacity to trigger the expression of the genes encoding TNF, IL-1b, IL-6 and IL-10 in RAW 264.7 macrophages. Immunophenotyping of PBMCs isolated from healthy individuals revealed that lipids from both BCG and <jats:italic>Mtb<\/jats:italic> were able to induce an increased frequency of CD4<jats:sup>+<\/jats:sup> and CD8<jats:sup>+<\/jats:sup> T cells, but only the lipid extract from <jats:italic>Mtb<\/jats:italic> enhanced the frequency of CD4<jats:sup>-<\/jats:sup>CD8<jats:sup>-<\/jats:sup>double-negative, \u03b3\u03c3<jats:sup>+<\/jats:sup>, CD4<jats:sup>+<\/jats:sup>HLA-DR<jats:sup>+<\/jats:sup>, and \u03b3\u03c3<jats:sup>+<\/jats:sup>HLA-DR<jats:sup>+<\/jats:sup> T cells relative to the nonstimulated control. Interestingly, only the <jats:italic>Mtb<\/jats:italic> lipid extract was able to increase the frequency of CD4<jats:sup>+<\/jats:sup> memory (CD45RO<jats:sup>+<\/jats:sup>) T cells, whereas the BCG lipid extract induced a diminished frequency of CD4<jats:sup>+<\/jats:sup> central memory (CD45RO<jats:sup>+<\/jats:sup>CCR7<jats:sup>-<\/jats:sup>) T cells after 48 h of culture compared to <jats:italic>Mtb<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>These findings show that the nonpolar lipids of the BCG bacilli presented diminished ability to trigger both proinflammatory and memory responses and suggest a potential use of <jats:italic>Mtb<\/jats:italic> lipids as adjuvants to increase the BCG vaccine efficacy.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2023.1263352","type":"journal-article","created":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T04:55:25Z","timestamp":1704948925000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Impaired macrophage and memory T-cell responses to Bacillus Calmette-Guerin nonpolar lipid extract"],"prefix":"10.3389","volume":"14","author":[{"given":"Alice","family":"Sarno","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Avelina","family":"Leite","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carlos","family":"Augusto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Igor","family":"Muller","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luanna","family":"de \u00c2ngelis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lilian","family":"Pimentel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adriano","family":"Queiroz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sergio","family":"Arruda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2024,1,11]]},"reference":[{"key":"B1","volume-title":"Global tuberculosis report 2022","year":"2022"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.3310\/hta17370","article-title":"Systematic review and meta-analysis of the current evidence on the duration of protection by bacillus Calmette\u2013Gu\u00e9rin vaccination against tuberculosis","volume":"17","author":"Abubakar","year":"2013","journal-title":"Health Technol Assess"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cit790","article-title":"Protection by BCG vaccine against tuberculosis: A systematic review of randomized controlled trials","volume":"58","author":"Mangtani","year":"2014","journal-title":"Clin Infect Dis"},{"key":"B4","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1136\/BMJ.G4643","article-title":"Effect of BCG vaccination against Mycobacterium tuberculosis infection in children: systematic review and meta-analysis","volume":"349","author":"Roy","year":"2014","journal-title":"BMJ"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-695X.2009.00559.x","article-title":"Comparable studies of immunostimulating activities in vitro among Mycobacterium bovis bacillus Calmette-Gu\u00e9rin (BCG) substrains","volume":"56","author":"Hayashi","year":"2009","journal-title":"FEMS Immunol Med Microbiol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2010.01947.x","article-title":"Biochemical characteristics among Mycobacterium bovis BCG substrains","volume":"306","author":"Hayashi","year":"2010","journal-title":"FEMS Microbiol Lett"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1315883111","article-title":"Molecular profiling of Mycobacterium tuberculosis identifies tuberculosinyl nucleoside products of the virulence-associated enzyme Rv3378c","volume":"111","author":"Layre","year":"2014","journal-title":"Proc Natl Acad Sci"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2016.18","article-title":"The BCG strain pool: diversity matters","volume":"24","author":"Bottai","year":"2016","journal-title":"Mol Ther"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0700869104","article-title":"Genome plasticity of BCG and impact on vaccine efficacy","volume":"104","author":"Brosch","year":"2007","journal-title":"Proc Natl Acad Sci"},{"key":"B10","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1590\/1678-4685-GMB-2021-0024","article-title":"In silico comparisons of lipid-related genes between Mycobacterium tuberculosis and BCG vaccine strains","volume":"44","author":"Sarno","year":"2021","journal-title":"Genet Mol Biol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1590\/0037-8682-0230-2016","article-title":"Bacterial immunostat: Mycobacterium tuberculosis lipids and their role in the host immune response","volume":"50","author":"Queiroz","year":"2017","journal-title":"Rev Soc Bras Med Trop"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.01848","article-title":"Differential host pro-inflammatory response to mycobacterial cell wall lipids regulated by the mce1 operon","volume":"11","author":"Petrilli","year":"2020","journal-title":"Front Immunol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2007.09.041","article-title":"Differential productions of lipid virulence factors among BCG vaccine strains and implications on BCG safety","volume":"25","author":"Chen","year":"2007","journal-title":"Vaccine"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/srep29076","article-title":"Loss of lipid virulence factors reduces the efficacy of the BCG vaccine","volume":"6","author":"Tran","year":"2016","journal-title":"Sci Rep"},{"key":"B15","doi-asserted-by":"publisher","first-page":"398","DOI":"10.1038\/MT.2015.216","article-title":"Variable virulence and efficacy of BCG vaccine strains in mice and correlation with genome polymorphisms","volume":"24","author":"Zhang","year":"2016","journal-title":"Mol Ther"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(13)60177-4","article-title":"Safety and efficacy of MVA85A, a new tuberculosis vaccine, in infants previously vaccinated with BCG: A randomised, placebo-controlled phase 2b trial","volume":"381","author":"Tameris","year":"2013","journal-title":"Lancet"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.tube.2013.01.003","article-title":"Lessons learnt from the first efficacy trial of a new infant tuberculosis vaccine since BCG","volume":"93","author":"Tameris","year":"2013","journal-title":"Tuberculosis"},{"key":"B18","doi-asserted-by":"publisher","first-page":"138","DOI":"10.1056\/NEJMOA1714021","article-title":"Prevention of infection with mycobacterium tuberculosis by H4:IC31\u00ae Vaccination or BCG revaccination in adolescents","volume":"379","author":"Nemes","year":"2018","journal-title":"N Engl J Med"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMOA1803484\/SUPPL_FILE\/NEJMOA1803484_DISCLOSURES.PDF","article-title":"Phase 2b controlled trial of M72\/AS01 E vaccine to prevent tuberculosis","volume":"379","author":"van der Meeren","year":"2018","journal-title":"New Engl J Med"},{"key":"B20","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1038\/s41586-019-1817-8","article-title":"Prevention of tuberculosis in nonhuman primates following intravenous BCG immunization","volume":"577","author":"Darrah","year":"2019","journal-title":"Nat Press"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-018-0319-9","article-title":"Prevention of tuberculosis infection and disease by local BCG in repeatedly exposed rhesus macaques","volume":"25","author":"Dijkman","year":"2019","journal-title":"Nat Med"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1038\/nature02837","article-title":"A glycolipid of hypervirulent tuberculosis strains that inhibits the innate immune response","volume":"431","author":"Reed","year":"2004","journal-title":"Nature"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2011.10.013","article-title":"A comparative lipidomics platform for Mycobacterium tuberculosis provides chemotaxonomic analysis for biomarker discovery","volume":"18","author":"Layre","year":"2011","journal-title":"Chem Biol"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1186\/1297-9716-43-54","article-title":"Differential effects of Mycobacterium bovis - Derived polar and apolar lipid fractions on bovine innate immune cells","volume":"43","author":"Pirson","year":"2012","journal-title":"Vet Res"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1006\/METH.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2013\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1038\/nm859","article-title":"Recombinant BCG exporting ESAT-6 confers enhanced protection against tuberculosis","volume":"9","author":"Pym","year":"2003","journal-title":"Nat Med"},{"key":"B27","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1186\/s12967-017-1301-x","article-title":"A lipidated peptide of Mycobacterium tuberculosis resuscitates the protective efficacy of BCG vaccine by evoking memory T cell immunity","volume":"15","author":"Rai","year":"2017","journal-title":"J Transl Med"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0045888","article-title":"Oral vaccination with lipid-formulated BCG induces a long-lived, multifunctional CD4+ T cell memory immune response","volume":"7","author":"Ancelet","year":"2012","journal-title":"PloS One"},{"key":"B29","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ebiom.2022.104023","article-title":"A CD4+ TNF+ monofunctional memory T-cell response to BCG vaccination is associated with Mycobacterium tuberculosis infection in infants exposed to HIV","volume":"80","author":"Warr","year":"2022","journal-title":"EBioMedicine"},{"key":"B30","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1371\/JOURNAL.PONE.0258743","article-title":"BCG vaccination induces enhanced frequencies of memory T cells and altered plasma levels of common \u03b3c cytokines in elderly individuals","volume":"16","author":"Kumar","year":"2021","journal-title":"PloS One"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/J.IMBIO.2010.09.003","article-title":"The immune responses of central and effector memory BCG-specific CD4+ T cells in BCG-vaccinated PPD+ donors were modulated by Treg cells","volume":"216","author":"Li","year":"2011","journal-title":"Immunobiology"},{"key":"B32","doi-asserted-by":"publisher","first-page":"618","DOI":"10.1128\/CVI.00368-09","article-title":"Phenotypic definition of effector and memory T-lymphocyte subsets in mice chronically infected with mycobacterium tuberculosis","volume":"17","author":"Henao-Tamayo","year":"2010","journal-title":"Clin Vaccine Immunol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.3389\/FIMMU.2020.566319\/BIBTEX","article-title":"Mycobacteria-specific T cells are generated in the lung during mucosal BCG immunization or infection with mycobacterium tuberculosis","volume":"11","author":"Basile","year":"2020","journal-title":"Front Immunol"},{"key":"B34","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/NCOMMS9533","article-title":"Mucosal vaccination with attenuated Mycobacterium tuberculosis induces strong central memory responses and protects against tuberculosis","volume":"6","author":"Kaushal","year":"2015","journal-title":"Nat Commun"},{"key":"B35","doi-asserted-by":"publisher","first-page":"1928","DOI":"10.1093\/INFDIS\/JIU347","article-title":"Central memory CD4+ T cells are responsible for the recombinant bacillus calmette-gu\u00e9rin \u0394ureC::hly vaccine\u2019s superior protection against tuberculosis","volume":"210","author":"Vogelzang","year":"2014","journal-title":"J Infect Dis"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1002\/EJI.201141903","article-title":"Protective immunity afforded by attenuated, PhoP-deficient Mycobacterium tuberculosis is associated with sustained generation of CD4+ T-cell memory","volume":"42","author":"Nambiar","year":"2012","journal-title":"Eur J Immunol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1263352\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T04:55:31Z","timestamp":1704948931000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1263352\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,11]]},"references-count":36,"alternative-id":["10.3389\/fimmu.2023.1263352"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2023.1263352","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,11]]},"article-number":"1263352"}}