A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model
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 interfer...
| Main Authors: | , , , , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
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Elsevier
2025
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| Online Access: | 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 |
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| author | 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_browse | Bigi, Fabiana Blanco, Federico Carlos Canal, Ana Granero, Gladys Klepp, Laura Ines Marini, María Rocío Onnainty, Renée Rocha, Rosana Valeria Vazquez, Cristina Lourdes Villafañe, Luciana María |
| 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_sort | Blanco, Federico Carlos |
| collection | INTA Digital |
| 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. |
| format | Artículo |
| id | INTA24780 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA247802025-12-29T10:03:10Z A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model 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 Vacuna Mycobacterium bovis Tuberculosis Bovina Vaccines Bovine Tuberculosis 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 2025-12-26T18:05:10Z 2025-12-26T18:05:10Z 2025-09 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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 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) application/pdf Elsevier Veterinary Immunology and Immunopathology 285 : 111001 (Septiembre 2025) |
| spellingShingle | Vacuna Mycobacterium bovis Tuberculosis Bovina Vaccines Bovine Tuberculosis 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 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 |
| 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_short | A DIVA-compatible Mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model |
| title_sort | diva compatible mycobacterium bovis triple mutant vaccine confers protection against bovine tuberculosis in mouse model |
| topic | Vacuna Mycobacterium bovis Tuberculosis Bovina Vaccines Bovine Tuberculosis |
| 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 |
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