Engineering a new vaccine platform for heterologous antigen delivery in live-attenuated Mycobacterium tuberculosis

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dc.contributor(LDV) Lab. Desenvolvimento de Vacinaspt_BR
dc.contributor.authorBroset, Estherpt_BR
dc.contributor.authorSeral, Juan Calvetpt_BR
dc.contributor.authorArnal, Carmenpt_BR
dc.contributor.authorUranga, Santiagopt_BR
dc.contributor.authorKanno, Alex Issamupt_BR
dc.contributor.authorLeite, Luciana Cezar de Cerqueirapt_BR
dc.contributor.authorMartín, Carlospt_BR
dc.contributor.authorGonzalo-Asensio, Jesúspt_BR
dc.identifier.citationBroset E, Seral JC, Arnal C, Uranga S, Kanno AI, Leite LCC, et al. Engineering a new vaccine platform for heterologous antigen delivery in live-attenuated Mycobacterium tuberculosis. Comput Struct Biotechnol J. 2021 July;19:4273-4283. doi:10.1016/j.csbj.2021.07.035.pt_BR
dc.description.abstractLive vaccines are attractive vehicles for antigen delivery as a strategy to immunize against heterologous pathogens. The live vaccine MTBVAC is based on rational attenuation of Mycobacterium tuberculosis with the objective of improving BCG protection against pulmonary tuberculosis. However, the development of recombinant mycobacteria as antigen-presenting microorganisms has been hindered due to their fastidious genetic manipulation. In this study, we used MTBVAC as a genetic platform to deliver diphtheria, tetanus, or pertussis toxoids, which are the immunogenic constituents of the DTP vaccine. When using nonoptimal genetic conditions, the expression of these immunogens was barely detectable. Accordingly, we pursued a rational, step-by-step optimization of the genetic components to achieve the expression and secretion of these toxoids. We explored variants of the L5 mycobacteriophage promoter to ensure balanced antigen expression and plasmid stability. Optimal signal sequences were identified by comparative proteomics of MTBVAC and its parental strain. It was determined that proteins secreted by the Twin Arginine Translocation pathway displayed higher secretion in MTBVAC, and the Ag85A secretion sequence was selected as the best candidate. Because the coding regions of diphtheria, tetanus, and pertussis toxoids significantly differ in G + C content relative to mycobacterial genes, their codon usage was optimized. We also placed a 3xFLAG epitope in frame with the C-terminus of these toxoids to facilitate protein detection. Altogether, these optimizations resulted in the secretion of DTP antigens by MTBVAC, as demonstrated by western blot and MRM-MS. Finally, we examined specific antibody responses in mice vaccinated with recombinant MTBVAC expressing DTP antigens.pt_BR
dc.description.sponsorship(MINECO) Spanish Ministry of Economy and Competitivenesspt_BR
dc.description.sponsorship(MCIN) Ministry for Science and Innovationpt_BR
dc.description.sponsorship(FEDER) El Fondo Europeo de Desarrollo Regionalpt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.relation.ispartofComputational and Structural Biotechnology Journalpt_BR
dc.rightsOpen accesspt_BR
dc.titleEngineering a new vaccine platform for heterologous antigen delivery in live-attenuated Mycobacterium tuberculosispt_BR
dc.rights.licenseCC BY-NC-NDpt_BR
dc.contributor.externalUniversidad de Zaragozapt_BR
dc.contributor.externalCarlos III Health Institutept_BR
dc.contributor.externalHospital Universitario Miguel Servetpt_BR
dc.contributor.external(BIFI) Instituto de Biocomputacion y Física de Sistemas Complejospt_BR
dc.subject.keywordSynthetic biologypt_BR
dc.subject.keywordFastidious bacteriapt_BR
dc.subject.keywordTuberculosis vaccinept_BR
dc.subject.keywordDTP antigenspt_BR
dc.subject.keywordTwin arginine translocation (TAT)pt_BR
dc.relation.ispartofabbreviatedComput Struct Biotechnol Jpt_BR
dc.identifier.citationabntv. 19, p. 4273-4283, jul. 2021pt_BR
dc.identifier.citationvancouver2021 July;19:4273-4283pt_BR
dc.contributor.butantanKanno, Alex Issamu|:Docente|:(LDV) Lab. Desenvolvimento de Vacinaspt_BR
dc.contributor.butantanLeite, Luciana Cezar de Cerqueira|:Pesquisador|:(LDV) Lab. Desenvolvimento de Vacinaspt_BR
dc.sponsorship.butantanSpanish Ministry of Economy and Competitiveness (MINECO)¦¦BES-2012-052937pt_BR
dc.sponsorship.butantanSpanish Ministry of Economy and Competitiveness (MINECO)¦¦BFU2015-72190-EXPpt_BR
dc.sponsorship.butantanMinistry for Science and Innovation (MCIN)¦¦RTI2018-097625-B-I00pt_BR
dc.sponsorship.butantanMinistry for Science and Innovation (MCIN)¦¦PID2019-104690RB-I00pt_BR
dc.sponsorship.butantanEl(FEDER) El Fondo Europeo de Desarrollo Regional¦¦2014-2020pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2017/24632-8pt_BR
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