المؤلفون: Marina A. Forrellad, Cristina L. Vázquez, Federico C. Blanco, Laura I. Klepp, Elizabeth A. García, Rosana V. Rocha, Villafañe Luciana, María M. Bigi, Maximiliano G. Gutierrez, Fabiana Bigi

المصدر: Virulence, Vol 10, Iss 1, Pp 1026-1033 (2019)

مصطلحات موضوعية: mycobacterium tuberculosis, oxidative stress, virulence factor, rv2617c, erp, Infectious and parasitic diseases, RC109-216

الوصف: In this study, we characterized the role of Rv2617c in the virulence of Mycobacterium tuberculosis. Rv2617c is a protein of unknown function unique to M. tuberculosis complex (MTC) and Mycobacterium leprae. In vitro, this protein interacts with the virulence factor P36 (also named Erp) and KdpF, a protein linked to nitrosative stress. Here, we showed that knockout of the Rv2617c gene in M. tuberculosis CDC1551 reduced the replication of the pathogen in a mouse model of infection and favored the trafficking of mycobacteria to phagolysosomes. We also demonstrated that Rv2617c and P36 are required for resistance to in vitro hydrogen peroxide treatment in M. tuberculosis and Mycobacterium bovis, respectively. These findings indicate Rv2617c and P36 act in concert to prevent bacterial damage upon oxidative stress.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2150-5594Test; https://doaj.org/toc/2150-5608Test

الوصول الحر: https://doaj.org/article/9240374c58df4e1ca84a4178b78e37bcTest

المؤلفون: Villafañe, Luciana, Rocha, Rosana Valeria, Bigi, María Mercedes, Klepp, Laura Inés, Taboga, Oscar Alberto, Forrellad, Marina Andrea, López, María Gabriela, Bigi, Fabiana

المصدر: Vet Immunol Immunopathol ; ISSN:1873-2534 ; Volume:273

مصطلحات موضوعية: Antigens, EsxG, IFN-γ assay, Mb0309, Mycobacterium bovis

الوصف: Bovine tuberculosis (bTB) represents a threat to livestock production. Mycobacterium bovis is the main causative agent of bTB and a pathogen capable of infecting wildlife and humans. Eradication programs based on surveillance in slaughterhouses with mandatory testing and culling of reactive cattle have failed to eradicate bTB in many regions worldwide. Therefore, developing effective tools to control this disease is crucial. Using a computational tool, we identified proteins in the M. bovis proteome that carry predictive binding peptides to BoLADRB3.2 and selected Mb0309, Mb1090, Mb1810 and Mb3810 from all the identified proteins. The expression of these proteins in a baculovirus-insect cell expression system was successful only for Mb0309 and Mb3810. In parallel, we expressed the ESAT-6 family proteins EsxG and EsxH in this system. Among the recombinant proteins, Mb0309 and EsxG exhibited moderate performance in distinguishing between cattle that test positive and negative to bTB using the official test, the intradermal tuberculin test (IDT), when used to stimulate interferon-gamma production in blood samples from cattle. However, when combined as a protein cocktail, Mb0309 and EsxG were reactive in 50 % of positive cattle. Further assessments in cattle that evade the IDT (false negative) and cattle infected with Mycobacterium avium paratuberculosis are necessary to determine the potential utility of this cocktail as an additional tool to assist the accurate diagnosis of bTB.

العلاقة: https://doi.org/10.1016/j.vetimm.2024.110788Test; https://pubmed.ncbi.nlm.nih.gov/38838485Test

الإتاحة: https://doi.org/10.1016/j.vetimm.2024.110788Test
https://pubmed.ncbi.nlm.nih.gov/38838485Test

المؤلفون: Villafañe, Luciana, Vaulet, Lucía Gallo, Viere, Florencia M., Klepp, Laura I., Forrellad, Marina A., Bigi, María M., Romano, María I., Magistrelli, Giovanni, Fermepin, Marcelo Rodríguez, Bigi, Fabiana

المساهمون: Agencia Nacional de Promoción Científica y Tecnológica, Instituto Nacional de Tecnología Agropecuaria

المصدر: Journal of Immunological Methods ; volume 500, page 113182 ; ISSN 0022-1759

الإتاحة: https://doi.org/10.1016/j.jim.2021.113182Test
https://api.elsevier.com/content/article/PII:S0022175921002271?httpAccept=text/xmlTest
https://api.elsevier.com/content/article/PII:S0022175921002271?httpAccept=text/plainTest

المؤلفون: Forrellad, Marina A., Vázquez, Cristina L., Blanco, Federico C., Klepp, Laura Inés, García, Elizabeth A., Rocha, Rosana V., Villafañe, Luciana, Bigi, María Mercedes

المصدر: Virulence ; Vol.10, no.1 ; 1026–1033 ; https://taylorandfrancis.comTest

مصطلحات موضوعية: MYCOBACTERIUM TUBERCULOSIS, OXIDATIVE STRESS, VIRULENCE FACTOR, RV2617C, ERP

الوصف: Fil: Forrellad, Marina A. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Hurlingham, Buenos Aires, Argentina. ; Fil: Vázquez, Cristina L. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Hurlingham, Buenos Aires, Argentina. ; Fil: Blanco, Federico C. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Hurlingham, Buenos Aires, Argentina. ; Fil: Klepp, Laura Inés. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Hurlingham, Buenos Aires, Argentina. ; Fil: García, Elizabeth A. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Hurlingham, Buenos Aires, Argentina. ; Fil: Rocha, Rosana V. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Hurlingham, Buenos Aires, Argentina. ; Fil: Villafañe, Luciana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Hurlingham, Buenos Aires, Argentina. ; Fil: Bigi, María Mercedes. Universidad de Buenos Aires. Facultad de Agronomía. Buenos Aires, Argentina. ; In this study, we characterized the role of Rv2617c in the virulence of Mycobacterium tuberculosis. Rv2617c is a protein of unknown function unique to M. tuberculosis complex (MTC) and Mycobacterium leprae. In vitro, this protein interacts with the virulence factor P36 (also named Erp) and KdpF, a protein linked to nitrosative stress. Here, we showed that knockout of the Rv2617c gene in M. tuberculosis CDC1551 reduced the replication of the pathogen in a mouse model of infection and favored the trafficking of mycobacteria to phagolysosomes. We also demonstrated that Rv2617c and P36 are required for resistance to in vitro hydrogen peroxide treatment in M. tuberculosis and Mycobacterium bovis, respectively. These findings indicate Rv2617c and P36 act in concert to prevent bacterial damage upon oxidative stress. ; grafs., fot.

وصف الملف: application/pdf

العلاقة: http://ri.agro.uba.ar/greenstone3/library/collection/arti/document/2019forrelladTest

الإتاحة: https://doi.org/10.1080/21505594.2019.1693714Test
http://ri.agro.uba.ar/greenstone3/library/collection/arti/document/2019forrelladTest

المؤلفون: Villafañe, Luciana, Forrellad, Marina Andrea, López, María Gabriela, Garbaccio, Sergio, Garro, Carlos, Rocha, Rosana Valeria, Eirin, María Emilia, Singh, Mahavir, Taboga, Oscar A., Bigi, Fabiana

المصدر: Microbial Physiology ; volume 29, issue 1-6, page 83-90 ; ISSN 2673-1665 2673-1673

مصطلحات موضوعية: Cell Biology, Applied Microbiology and Biotechnology, Physiology, Biochemistry, Microbiology, Biotechnology

الوصف: Bovine tuberculosis (bTB) is a disease produced by Mycobacterium bovis that affects livestock, wild animals, and humans. The classical diagnostic method to detect bTB is measuring the response induced with the intradermal injection of purified protein derivative of M. bovis (PPDb). Another ancillary bTB test detects IFN-γ produced in whole blood upon stimulation with PPDb, protein/peptide cocktails, or individual antigens. Among the most used M. bovis antigens in IFN-γ assays are the secreted proteins ESAT-6 and CFP-10, which together with antigen Rv3615c improve the sensitivity of the test in comparison to PPDb. Protein reagents for immune stimulation are generally obtained from Escherichia coli , because this bacterium produces a high level of recombinant proteins. However, E. coli recombinant antigens are in general contaminated with lipopolysaccharides and other components that produce non-specific IFN-γ secretion in in vitro assays. In this work, we produced the relevant ESAT-6, CFP-10, and Rv3615c M. bovis antigens as fusions to the polyhedrin protein from the baculovirus AcMNPV. We obtained chimeric proteins effectively incorporated to the occlusion bodies and easily purified the recombinant polyhedra with no reactive contaminants. In an IFN-γ assay, these fusion proteins showed equivalent sensibility but better specificity than the same M. bovis proteins produced in E. coli .

الإتاحة: https://doi.org/10.1159/000506687Test
https://www.karger.com/Article/Pdf/506687Test

المؤلفون: Villafañe, Luciana, Forrellad, Marina Andrea, López, María Gabriela, Garbaccio, Sergio, Garro, Carlos, Rocha, Rosana Valeria, Eirin, María Emilia, Singh, Mahavir, Taboga, Oscar A., Bigi, Fabiana

المصدر: Journal of Molecular Microbiology and Biotechnology; July 2020, Vol. 29 Issue: 1-6 p83-90, 8p

مستخلص: Bovine tuberculosis (bTB) is a disease produced by Mycobacterium bovisthat affects livestock, wild animals, and humans. The classical diagnostic method to detect bTB is measuring the response induced with the intradermal injection of purified protein derivative of M. bovis(PPDb). Another ancillary bTB test detects IFN-γ produced in whole blood upon stimulation with PPDb, protein/peptide cocktails, or individual antigens. Among the most used M. bovisantigens in IFN-γ assays are the secreted proteins ESAT-6 and CFP-10, which together with antigen Rv3615c improve the sensitivity of the test in comparison to PPDb. Protein reagents for immune stimulation are generally obtained from Escherichia coli, because this bacterium produces a high level of recombinant proteins. However, E. colirecombinant antigens are in general contaminated with lipopolysaccharides and other components that produce non-specific IFN-γ secretion in in vitro assays. In this work, we produced the relevant ESAT-6, CFP-10, and Rv3615c M. bovisantigens as fusions to the polyhedrin protein from the baculovirus AcMNPV. We obtained chimeric proteins effectively incorporated to the occlusion bodies and easily purified the recombinant polyhedra with no reactive contaminants. In an IFN-γ assay, these fusion proteins showed equivalent sensibility but better specificity than the same M. bovisproteins produced in E. coli.

المؤلفون: Villafañe, Luciana María, Vaulet, Lucía Gallo, Viere, Florencia M., Klepp, Laura Ines, Forrellad, Marina Andrea, Pujadas Bigi, María Montserrat, Romano, Maria Isabel, Magistrelli, Giovanni, Rodríguez Fermepin, Marcelo, Bigi, Fabiana

مصطلحات موضوعية: COVID-19, ELISA, RBD, SARS-COV-2, SEROLOGICAL TEST, https://purl.org/becyt/ford/1.6Test, https://purl.org/becyt/ford/1Test

الوصف: Serology tests for SARS-CoV-2 have proven to be important tools to fight against the COVID-19 pandemic. These serological tests can be used in low-income and remote areas for patient contact tracing, epidemiologic studies and vaccine efficacy evaluations. In this study, we used a semi-stable mammalian episomal expression system to produce high quantities of the receptor-binding domain-RBD of SARS-CoV-2 in a simple and very economical way. The recombinant antigen was tested in an in-house IgG ELISA for COVID-19 with a panel of human sera. A performance comparison of this serology test with a commercial test based on the full-length spike protein showed 100% of concordance between tests. Thus, this serological test can be an attractive and inexpensive option in scenarios of limited resources to face the COVID-19 pandemic. ; Fil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina ; Fil: Vaulet, Lucía Gallo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Fisiopatología y Bioquímica Clínica; Argentina ; Fil: Viere, Florencia M. Universidad de Buenos Aires. Facultad de Medicina; Argentina ; Fil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina ; Fil: Forrellad, Marina Andrea. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de ...

وصف الملف: application/pdf

العلاقة: info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0022175921002271Test; http://hdl.handle.net/11336/163019Test; Villafañe, Luciana María; Vaulet, Lucía Gallo; Viere, Florencia M.; Klepp, Laura Ines; Forrellad, Marina Andrea; et al.; Development and evaluation of a low cost IgG ELISA test based in RBD protein for COVID-19; Elsevier Science; Journal Of Immunological Methods; 500; 11-2021; 1-5; CONICET Digital; CONICET

الإتاحة: https://doi.org/10.1016/j.jim.2021.113182Test
http://hdl.handle.net/11336/163019Test

المؤلفون: Villafañe, Luciana María, Forrellad, Marina Andrea, López, María Gabriela, Garbaccio, Sergio Gabriel, Garro, Carlos, Rocha, Rosana Valeria, Eirin, Maria Emilia, Singh, Mahavir, Taboga, Oscar Alberto, Bigi, Fabiana

مصطلحات موضوعية: ESAT-6, CFP10, RV3615c, MYCOBACTERIUM BOVIS, OCCLUSION BODIES, BACULOVIRUS, IFN-γ ASSAY, https://purl.org/becyt/ford/1.6Test, https://purl.org/becyt/ford/1Test

الوصف: Bovine tuberculosis (bTB) is a disease produced by Mycobacterium bovis that affects livestock, wild animals, and humans. The classical diagnostic method to detect bTB is measuring the response induced with the intradermal injection of purified protein derivative of M. bovis (PPDb). Another ancillary bTB test detects IFN-γproduced in whole blood upon stimulation with PPDb, protein/peptide cocktails, or individual antigens. Among the most used M. bovis antigens in IFN-γassays are the secreted proteins ESAT-6 and CFP-10, which together with antigen Rv3615c improve the sensitivity of the test in comparison to PPDb. Protein reagents for immune stimulation are generally obtained from Escherichia coli, because this bacterium produces a high level of recombinant proteins. However, E. coli recombinant antigens are in general contaminated with lipopolysaccharides and other components that produce non-specific IFN-γsecretion in in vitro assays. In this work, we produced the relevant ESAT-6, CFP-10, and Rv3615c M. bovis antigens as fusions to the polyhedrin protein from the baculovirus AcMNPV. We obtained chimeric proteins effectively incorporated to the occlusion bodies and easily purified the recombinant polyhedra with no reactive contaminants. In an IFN-γassay, these fusion proteins showed equivalent sensibility but better specificity than the same M. bovis proteins produced in E. coli. ; Fil: Villafañe, Luciana María. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina ; Fil: Forrellad, Marina Andrea. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de ...

وصف الملف: application/pdf

العلاقة: info:eu-repo/semantics/altIdentifier/url/https://www.karger.com/Article/FullText/506687Test; http://hdl.handle.net/11336/142408Test; Villafañe, Luciana María; Forrellad, Marina Andrea; López, María Gabriela; Garbaccio, Sergio Gabriel; Garro, Carlos; et al.; Production of Mycobacterium bovis Antigens Included in Recombinant Occlusion Bodies of Baculovirus; Karger; Journal of Molecular Microbiology and Biotechnology; 29; 1-6; 7-2020; 83-90; CONICET Digital; CONICET

الإتاحة: https://doi.org/10.1159/000506687Test
http://hdl.handle.net/11336/142408Test

المؤلفون: Forrellad, Marina A., Vázquez, Cristina L., Blanco, Federico C., Klepp, Laura I., García, Elizabeth A., Rocha, Rosana V., Villafañe Luciana, Bigi, María M., Gutierrez, Maximiliano G., Bigi, Fabiana

مصطلحات موضوعية: 3. Good health

الوصف: In this study, we characterized the role of Rv2617c in the virulence of Mycobacterium tuberculosis. Rv2617c is a protein of unknown function unique to M. tuberculosis complex (MTC) and Mycobacterium leprae. In vitro, this protein interacts with the virulence factor P36 (also named Erp) and KdpF, a protein linked to nitrosative stress. Here, we showed that knockout of the Rv2617c gene in M. tuberculosis CDC1551 reduced the replication of the pathogen in a mouse model of infection and favored the trafficking of mycobacteria to phagolysosomes. We also demonstrated that Rv2617c and P36 are required for resistance to in vitro hydrogen peroxide treatment in M. tuberculosis and Mycobacterium bovis, respectively. These findings indicate Rv2617c and P36 act in concert to prevent bacterial damage upon oxidative stress.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::6729b67b9c1d78ad9a2f15cd7046f2acTest

المؤلفون: Forrellad, Marina A, Vázquez, Cristina L, Blanco, Federico C, Klepp, Laura I, García, Elizabeth A, Rocha, Rosana V, Villafañe Luciana, Bigi, María M, Gutierrez, Maximiliano G, Bigi, Fabiana

مصطلحات موضوعية: Model organisms, Infectious Disease, Cell Biology, 3. Good health

الوصف: In this study, we characterized the role of Rv2617c in the virulence of Mycobacterium tuberculosis. Rv2617c is a protein of unknown function unique to M. tuberculosis complex (MTC) and Mycobacterium leprae. In vitro, this protein interacts with the virulence factor P36 (also named Erp) and KdpF, a protein linked to nitrosative stress. Here, we showed that knockout of the Rv2617c gene in M. tuberculosis CDC1551 reduced the replication of the pathogen in a mouse model of infection and favored the trafficking of mycobacteria to phagolysosomes. We also demonstrated that Rv2617c and P36 are required for resistance to in vitro hydrogen peroxide treatment in M. tuberculosis and Mycobacterium bovis, respectively. These findings indicate Rv2617c and P36 act in concert to prevent bacterial damage upon oxidative stress.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::5bc50beb023585be69be5a710563aeeaTest