A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model

Autores
Blanco, Federico Carlos; Onnainty, Renée; Marini, María Rocío; Klepp, Laura Ines; Rocha, Rosana Valeria; Villafañe, Luciana María; Vazquez, Cristina Lourdes; Canal, Ana; Granero, Gladys; Bigi, Fabiana
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Bovine tuberculosis (bTB) is a pulmonary infectious disease caused by Mycobacterium bovis, affecting cattle and a wide range of mammals, including humans. Despite its significant impact on global livestock production, no commercial vaccine is currently available, partly due to potential interference with standard diagnostic tests. In this study, we evaluated the protective efficacy of a triple M. bovis mutant lacking the immunodominant antigens ESAT-6 and CFP-10, as well as the virulence factor Ag85A. This mutant is compatible with DIVA (Differentiation of Infected from Vaccinated Animals) diagnostics based on ESAT-6 and CFP-10 detection. The triple mutant was assayed both alone and in a heterologous prime-boost regimen using recombinant Ag85A conjugated to chitosan nanocapsules. Protection was assessed by quantifying M. bovis colony-forming units (CFUs) in the lungs and spleen following challenge. Organ homogenates were cultured on solid media, and CFUs were enumerated at five and ten weeks post-plating. At five weeks, all vaccinated groups demonstrated comparable protection in the lungs. In the spleen, both the triple mutant and BCG groups showed reduced CFU counts compared to the un vaccinated group. By ten weeks, lung protection was most pronounced in the prime-boost and BCG groups, whereas spleen protection was restricted to the prime-boost group. At this stage, persistence of the triple mutant was detected in both lungs and spleen, highlighting the need for further evaluation of its residual virulence. Post- challenge immune responses were assessed by measuring CD4 +KLRG1-CXCL3 + T cells, a subset previously associated with protective immunity against tuberculosis, among other T cell populations evaluated. Vaccinated mice exhibited a significant expansion of this population compared to unvaccinated controls. Notably, higher frequencies of these cells correlated with reduced pulmonary bacterial burden, reinforcing their potential as a biomarker of protective immunity.
Instituto de Biotecnología
Fil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Blanco, Federico Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Onnainty, Renée. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Ciencias Farmacéuticas. Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Onnainty, Renée. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Marini, María Rocío. Universidad Nacional del Litoral (UNL). Facultad de Ciencias Veterinarias. Laboratorio de Anatomía Patológica; Argentina
Fil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Klepp, Laura Ines. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Rocha, Rosana Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Rocha, Rosana Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Rocha, Rosana Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Villafañe, Luciana María. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO) ; Argentina
Fil: Vazquez, Cristina Lourdes. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Vazquez, Cristina Lourdes. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Vazquez, Cristina Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Canal, Ana. Universidad Nacional del Litoral (UNL). Facultad de Ciencias Veterinarias. Laboratorio de Anatomía Patológica; Argentina
Fil: Granero, Gladys. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Ciencias Farmacéuticas. Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Granero, Gladys. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Bigi, Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fuente
Veterinary Immunology and Immunopathology 285 : 111001 (Septiembre 2025)
Materia
Vacuna
Mycobacterium bovis
Tuberculosis Bovina
Vaccines
Bovine Tuberculosis
Nivel de accesibilidad
acceso restringido
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/24780
Acceder
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oai_identifier_str oai:localhost:20.500.12123/24780
network_acronym_str INTADig
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network_name_str INTA Digital (INTA)
spelling A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse modelBlanco, Federico CarlosOnnainty, RenéeMarini, María RocíoKlepp, Laura InesRocha, Rosana ValeriaVillafañe, Luciana MaríaVazquez, Cristina LourdesCanal, AnaGranero, GladysBigi, FabianaVacunaMycobacterium bovisTuberculosis BovinaVaccinesBovine TuberculosisBovine tuberculosis (bTB) is a pulmonary infectious disease caused by Mycobacterium bovis, affecting cattle and a wide range of mammals, including humans. Despite its significant impact on global livestock production, no commercial vaccine is currently available, partly due to potential interference with standard diagnostic tests. In this study, we evaluated the protective efficacy of a triple M. bovis mutant lacking the immunodominant antigens ESAT-6 and CFP-10, as well as the virulence factor Ag85A. This mutant is compatible with DIVA (Differentiation of Infected from Vaccinated Animals) diagnostics based on ESAT-6 and CFP-10 detection. The triple mutant was assayed both alone and in a heterologous prime-boost regimen using recombinant Ag85A conjugated to chitosan nanocapsules. Protection was assessed by quantifying M. bovis colony-forming units (CFUs) in the lungs and spleen following challenge. Organ homogenates were cultured on solid media, and CFUs were enumerated at five and ten weeks post-plating. At five weeks, all vaccinated groups demonstrated comparable protection in the lungs. In the spleen, both the triple mutant and BCG groups showed reduced CFU counts compared to the un vaccinated group. By ten weeks, lung protection was most pronounced in the prime-boost and BCG groups, whereas spleen protection was restricted to the prime-boost group. At this stage, persistence of the triple mutant was detected in both lungs and spleen, highlighting the need for further evaluation of its residual virulence. Post- challenge immune responses were assessed by measuring CD4 +KLRG1-CXCL3 + T cells, a subset previously associated with protective immunity against tuberculosis, among other T cell populations evaluated. Vaccinated mice exhibited a significant expansion of this population compared to unvaccinated controls. Notably, higher frequencies of these cells correlated with reduced pulmonary bacterial burden, reinforcing their potential as a biomarker of protective immunity.Instituto de BiotecnologíaFil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Blanco, Federico Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Onnainty, Renée. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Ciencias Farmacéuticas. Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); ArgentinaFil: Onnainty, Renée. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); ArgentinaFil: Marini, María Rocío. Universidad Nacional del Litoral (UNL). Facultad de Ciencias Veterinarias. Laboratorio de Anatomía Patológica; ArgentinaFil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Klepp, Laura Ines. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Rocha, Rosana Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Rocha, Rosana Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Rocha, Rosana Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Villafañe, Luciana María. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO) ; ArgentinaFil: Vazquez, Cristina Lourdes. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Vazquez, Cristina Lourdes. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Vazquez, Cristina Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Canal, Ana. Universidad Nacional del Litoral (UNL). Facultad de Ciencias Veterinarias. Laboratorio de Anatomía Patológica; ArgentinaFil: Granero, Gladys. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Ciencias Farmacéuticas. Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); ArgentinaFil: Granero, Gladys. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); ArgentinaFil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Bigi, Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaElsevier2025-12-26T18:05:10Z2025-12-26T18:05:10Z2025-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/24780https://www.sciencedirect.com/science/article/abs/pii/S01652427250012171873-25340165-2427https://doi.org/10.1016/j.vetimm.2025.111001Veterinary Immunology and Immunopathology 285 : 111001 (Septiembre 2025)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/2023-PD-L06-I116, Implementación de tecnologías y nuevas estrategias preventivas y terapéuticas para el desarrollo sustentable y eficiente de la producción animal en el marco de Una Saludinfo:eu-repo/semantics/restrictedAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2026-01-08T10:41:01Zoai:localhost:20.500.12123/24780instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2026-01-08 10:41:01.567INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
title A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
spellingShingle A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
Blanco, Federico Carlos
Vacuna
Mycobacterium bovis
Tuberculosis Bovina
Vaccines
Bovine Tuberculosis
title_short A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
title_full A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
title_fullStr A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
title_full_unstemmed A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
title_sort A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
dc.creator.none.fl_str_mv Blanco, Federico Carlos
Onnainty, Renée
Marini, María Rocío
Klepp, Laura Ines
Rocha, Rosana Valeria
Villafañe, Luciana María
Vazquez, Cristina Lourdes
Canal, Ana
Granero, Gladys
Bigi, Fabiana
author Blanco, Federico Carlos
author_facet Blanco, Federico Carlos
Onnainty, Renée
Marini, María Rocío
Klepp, Laura Ines
Rocha, Rosana Valeria
Villafañe, Luciana María
Vazquez, Cristina Lourdes
Canal, Ana
Granero, Gladys
Bigi, Fabiana
author_role author
author2 Onnainty, Renée
Marini, María Rocío
Klepp, Laura Ines
Rocha, Rosana Valeria
Villafañe, Luciana María
Vazquez, Cristina Lourdes
Canal, Ana
Granero, Gladys
Bigi, Fabiana
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Vacuna
Mycobacterium bovis
Tuberculosis Bovina
Vaccines
Bovine Tuberculosis
topic Vacuna
Mycobacterium bovis
Tuberculosis Bovina
Vaccines
Bovine Tuberculosis
dc.description.none.fl_txt_mv Bovine tuberculosis (bTB) is a pulmonary infectious disease caused by Mycobacterium bovis, affecting cattle and a wide range of mammals, including humans. Despite its significant impact on global livestock production, no commercial vaccine is currently available, partly due to potential interference with standard diagnostic tests. In this study, we evaluated the protective efficacy of a triple M. bovis mutant lacking the immunodominant antigens ESAT-6 and CFP-10, as well as the virulence factor Ag85A. This mutant is compatible with DIVA (Differentiation of Infected from Vaccinated Animals) diagnostics based on ESAT-6 and CFP-10 detection. The triple mutant was assayed both alone and in a heterologous prime-boost regimen using recombinant Ag85A conjugated to chitosan nanocapsules. Protection was assessed by quantifying M. bovis colony-forming units (CFUs) in the lungs and spleen following challenge. Organ homogenates were cultured on solid media, and CFUs were enumerated at five and ten weeks post-plating. At five weeks, all vaccinated groups demonstrated comparable protection in the lungs. In the spleen, both the triple mutant and BCG groups showed reduced CFU counts compared to the un vaccinated group. By ten weeks, lung protection was most pronounced in the prime-boost and BCG groups, whereas spleen protection was restricted to the prime-boost group. At this stage, persistence of the triple mutant was detected in both lungs and spleen, highlighting the need for further evaluation of its residual virulence. Post- challenge immune responses were assessed by measuring CD4 +KLRG1-CXCL3 + T cells, a subset previously associated with protective immunity against tuberculosis, among other T cell populations evaluated. Vaccinated mice exhibited a significant expansion of this population compared to unvaccinated controls. Notably, higher frequencies of these cells correlated with reduced pulmonary bacterial burden, reinforcing their potential as a biomarker of protective immunity.
Instituto de Biotecnología
Fil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Blanco, Federico Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Onnainty, Renée. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Ciencias Farmacéuticas. Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Onnainty, Renée. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Marini, María Rocío. Universidad Nacional del Litoral (UNL). Facultad de Ciencias Veterinarias. Laboratorio de Anatomía Patológica; Argentina
Fil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Klepp, Laura Ines. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Rocha, Rosana Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Rocha, Rosana Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Rocha, Rosana Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Villafañe, Luciana María. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO) ; Argentina
Fil: Vazquez, Cristina Lourdes. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Vazquez, Cristina Lourdes. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Vazquez, Cristina Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Canal, Ana. Universidad Nacional del Litoral (UNL). Facultad de Ciencias Veterinarias. Laboratorio de Anatomía Patológica; Argentina
Fil: Granero, Gladys. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Ciencias Farmacéuticas. Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Granero, Gladys. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Unidad de Investigaciones y Desarrollo en Tecnología Farmacéutica (UNITEFA); Argentina
Fil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Bigi, Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
description Bovine tuberculosis (bTB) is a pulmonary infectious disease caused by Mycobacterium bovis, affecting cattle and a wide range of mammals, including humans. Despite its significant impact on global livestock production, no commercial vaccine is currently available, partly due to potential interference with standard diagnostic tests. In this study, we evaluated the protective efficacy of a triple M. bovis mutant lacking the immunodominant antigens ESAT-6 and CFP-10, as well as the virulence factor Ag85A. This mutant is compatible with DIVA (Differentiation of Infected from Vaccinated Animals) diagnostics based on ESAT-6 and CFP-10 detection. The triple mutant was assayed both alone and in a heterologous prime-boost regimen using recombinant Ag85A conjugated to chitosan nanocapsules. Protection was assessed by quantifying M. bovis colony-forming units (CFUs) in the lungs and spleen following challenge. Organ homogenates were cultured on solid media, and CFUs were enumerated at five and ten weeks post-plating. At five weeks, all vaccinated groups demonstrated comparable protection in the lungs. In the spleen, both the triple mutant and BCG groups showed reduced CFU counts compared to the un vaccinated group. By ten weeks, lung protection was most pronounced in the prime-boost and BCG groups, whereas spleen protection was restricted to the prime-boost group. At this stage, persistence of the triple mutant was detected in both lungs and spleen, highlighting the need for further evaluation of its residual virulence. Post- challenge immune responses were assessed by measuring CD4 +KLRG1-CXCL3 + T cells, a subset previously associated with protective immunity against tuberculosis, among other T cell populations evaluated. Vaccinated mice exhibited a significant expansion of this population compared to unvaccinated controls. Notably, higher frequencies of these cells correlated with reduced pulmonary bacterial burden, reinforcing their potential as a biomarker of protective immunity.
publishDate 2025
dc.date.none.fl_str_mv 2025-12-26T18:05:10Z
2025-12-26T18:05:10Z
2025-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/24780
https://www.sciencedirect.com/science/article/abs/pii/S0165242725001217
1873-2534
0165-2427
https://doi.org/10.1016/j.vetimm.2025.111001
url http://hdl.handle.net/20.500.12123/24780
https://www.sciencedirect.com/science/article/abs/pii/S0165242725001217
https://doi.org/10.1016/j.vetimm.2025.111001
identifier_str_mv 1873-2534
0165-2427
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/2023-PD-L06-I116, Implementación de tecnologías y nuevas estrategias preventivas y terapéuticas para el desarrollo sustentable y eficiente de la producción animal en el marco de Una Salud
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv restrictedAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Veterinary Immunology and Immunopathology 285 : 111001 (Septiembre 2025)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
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instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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