Project description:The BCG vaccine (Bacille Calmette-Guerin), only prophylactic measure against tuberculosis (TB), was obtained in the early twentieth century by Calmette and Guérin after 231 passages of a M. bovis clinical isolate in medium containing glycerin and bovine bile. Its protective efficacy against pulmonary TB in adults varies from 0-80% and the genetic differences among vaccines strains used worldwide contribute to this variation. The Brazilian vaccine strain, BCG Moreau, is considered a primitive strain and more immunogenic, closer to the original BCG when compared to newer strains, such as BCG Pasteur (reference strain). The characterization of BCG sub-strains can contribute not only to a better understanding of the vaccine and its protective effect, but also, in elucidating how different BCG culture conditions may contribute to the impact on the host's immune response. Thus, we aimed to characterize the differences in gene expression through the intracellular proteomic profile of BCG Moreau and Pasteur strains, cultivated in Sauton or 7H9 medium, using the methodology of two-dimensional electrophoresis (2DE) and mass spectrometry.
Project description:Tuberculosis (TB) is one of major causes of death worldwide. Bacillus Calmette-Guerin (BCG) is the only licensed TB vaccine and its inability to protect against adult pulmonary TB can be due to genetic differences among strains described since the 1940s. In this work, we compared the proteomic profile of the surface-associated proteins from M. bovis BCG Moreau, the Brazilian vaccine strain, and the BCG Pasteur reference strain. The methodology used was 2D-gel electrophoresis combined with mass spectrometry techniques (MALDI-TOF/TOF). We identified 115 proteins. Of these, 24 proteins showed differential expression between the two BCG strains. Furthermore, 27 proteins previously described as displaying moonlighting function were identified, 8 of these proteins showed variation in abundance comparing BCG Moreau to Pasteur and 2 of them presented two different domain hits.