Proteomic profile of culture filtrate from the Brazilian vaccine strain Mycobacterium bovis BCG Moreau compared to M. bovis BCG Pasteur

<p>Abstract</p> <p>Background</p> <p>Bacille Calmette-Guerin (BCG) is currently the only available vaccine against tuberculosis (TB) and comprises a heterogeneous family of sub-strains with genotypic and phenotypic differences. The World Health Organization (WHO) affirms that the characterization of BCG sub-strains, both on genomic and proteomic levels, is crucial for a better comprehension of the vaccine. In addition, these studies can contribute in the development of a more efficient vaccine against TB. Here, we combine two-dimensional electrophoresis (2DE) and mass spectrometry to analyse the proteomic profile of culture filtrate proteins (CFPs) from <it>M. bovis </it>BCG Moreau, the Brazilian vaccine strain, comparing it to that of BCG Pasteur. CFPs are considered of great importance given their dominant immunogenicity and role in pathogenesis, being available for interaction with host cells since early infection.</p> <p>Results</p> <p>The 2DE proteomic map of <it>M. bovis </it>BCG Moreau CFPs in the pH range 3 - 8 allowed the identification of 158 spots corresponding to 101 different proteins, identified by MS/MS. Comparison to BCG Pasteur highlights the great similarity between these BCG strains. However, quantitative analysis shows a higher expression of immunogenic proteins such as Rv1860 (BCG1896, Apa), Rv1926c (BCG1965c, Mpb63) and Rv1886c (BCG1923c, Ag85B) in BCG Moreau when compared to BCG Pasteur, while some heat shock proteins, such as Rv0440 (BCG0479, GroEL2) and Rv0350 (BCG0389, DnaK), show the opposite pattern.</p> <p>Conclusions</p> <p>Here we report the detailed 2DE profile of CFPs from <it>M. bovis </it>BCG Moreau and its comparison to BCG Pasteur, identifying differences that may provide relevant information on vaccine efficacy. These findings contribute to the detailed characterization of the Brazilian vaccine strain against TB, revealing aspects that may lead to a better understanding of the factors leading to BCG's variable protective efficacy against TB.</p

Exploring the "Latin American Mediterranean" family and the RDRio lineage in Mycobacterium tuberculosis isolates from Paraguay, Argentina and Venezuela

Submitted by Sandra Infurna ([email protected]) on 2020-03-28T17:15:58Z No. of bitstreams: 1 HarrrisonGomes_SidraVasconcelos_etal_IOC_2019.pd.pdf: 1746618 bytes, checksum: 5564ed65983e850073abda10c0053dea (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2020-03-28T17:43:14Z (GMT) No. of bitstreams: 1 HarrrisonGomes_SidraVasconcelos_etal_IOC_2019.pd.pdf: 1746618 bytes, checksum: 5564ed65983e850073abda10c0053dea (MD5)Made available in DSpace on 2020-03-28T17:43:14Z (GMT). No. of bitstreams: 1 HarrrisonGomes_SidraVasconcelos_etal_IOC_2019.pd.pdf: 1746618 bytes, checksum: 5564ed65983e850073abda10c0053dea (MD5) Previous issue date: 2019Universidad Nacional de Asunción. Instituto de Investigaciones en Ciencias de la Salud. Departamento de Biología Molecular y Biotecnología. Asunción, Paraguay / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular aplicada às Micobactérias. Rio de Janeiro, RJ, Brasil.Universidad Nacional de Asunción. Instituto de Investigaciones en Ciencias de la Salud. Departamento de Biología Molecular y Biotecnología. Asunción, Paraguay.Universidad Nacional de Asunción. Instituto de Investigaciones en Ciencias de la Salud. Departamento de Biología Molecular y Biotecnología. Asunción, Paraguay.Instituto Nacional de Enfermedades Infecciosas, ANLIS “Carlos G. Malbran”. Servicio de Micobacterias. Buenos Aires. Argentina.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular aplicada às Micobactérias. Rio de Janeiro, RJ, Brasil..Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Microbiologia Celular, Rio de Janeiro, RJ, Brasil.Laboratorio Central de Salud Pública, MSP y BS. Asunción, Paraguay.Instituto Nacional de Enfermedades Respiratorias Emilio Coni. Buenos Aires, Argentina.Instituto de Biomedicina. Laboratorio de Tuberculosis. Caracas, Venezuela / Universidad de Las Américas Facultad de Ciencias de la Salud. One Health Research Group. Quito, Ecuador.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular aplicada às Micobactérias. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular aplicada às Micobactérias. Rio de Janeiro, RJ, Brasil.The Latin American & Mediterranean (LAM) spoligotype family is one of the most successful genotype of Mycobacterium tuberculosis worldwide and particularly prevalent in South-America. Within this family, a sublineage named Region of Difference Rio (RDRio) was reported initially in Brazil and is characterized by a genomic deletion of about 26.3 kb. This lineage seems to show a specific adaptation to the Euro-Latin American population. In this context, we sought to evaluate the LAM family and the presence of the RDRio genotype in samples from three Latin American countries including Paraguay, Venezuela and Argentina. To detect LAM strains reliably we applied a typing scheme using spoligotyping, 12 loci MIRU-VNTR, the Ag85C103 SNP and the regions of difference RDRio and RD174. IS6110-RFLP results were also used when available

Kinetoplastid membrane protein-11 exacerbates infection with Leishmania amazonensis in murine macrophages

In Leishmania amazonensis, kinetoplastid membrane protein-11 (KMP-11) expression increases during metacyclogenesis and is higher in amastigotes than in promastigotes, suggesting a role for this protein in the infection of the mammalian host. We show that the addition of KMP-11 exacerbates L. amazonensis infection in peritoneal macrophages from BALB/c mice by increasing interleukin (IL)-10 secretion and arginase activity while reducing nitric oxide (NO) production. The doses of KMP-11, the IL-10 levels and the intracellular amastigote loads were strongly, positively and significantly correlated. The increase in parasite load induced by KMP-11 was inhibited by anti-KMP-11 or anti-IL-10 neutralising antibodies, but not by isotype controls. The neutralising antibodies, but not the isotype controls, were also able to significantly decrease the parasite load in macrophages cultured without the addition of KMP-11, demonstrating that KMP-11-induced exacerbation of the infection is not dependent on the addition of exogenous KMP-11 and that the protein naturally expressed by the parasite is able to promote it. In this study, the exacerbating effect of KMP-11 on macrophage infection with Leishmania is for the first time demonstrated, implicating it as a virulence factor in L. amazonensis. The stimulation of IL-10 production and arginase activity and the inhibition of NO synthesis are likely involved in this effect