Project description:A lot of attempts have been made to understand the immunopathological mechanisms of Brucella canis infection because of the importance of the disease in both public and clinical aspects. However, previous mechanisms are not still revealed. Therefore, in vitro models, which mimic to in vivo infection route using a canine epithelial cell, D17 cell, and a canine macrophage, DH82 cell, was used to solve the clues by analysis of transcriptomes in the cells. In this study, a co-culture model was constructed using the two cells, D17 and DH82 cell lines with trans-well plate. Also, a single cell culture system using DH82 was established. After stimulation of the cells in two different systems with B. canis, gene expressions in the macrophages of the two different system were analyzed by RNA-sequencing.
Project description:Leishmania (L.) infantum is the etiologic agent of visceral leishmaniasis (VL). In Brazil represents a serious public health problem. Studies have shown that regulation of immune response appears to depend on miRNAs. In canine VL due to cell immune suppression being determinant of disease progression, knowledge of miRNAs may be important for the pattern of change in immune response. Here, we suggest that post-transcriptional regulation, mediated by miRNAs, may play a role in immune response of dogs with VL.
Project description:Gram-negative bacteria release nanovesicles, called outer membrane vesicles (OMVs), from their outer membrane. Proteomics has been used to determine their composition. OMVs contain proteins able to elicit an immune response, so they have been proposed as a model to develop acellular vaccines. In this study, OMVs of Brucella suis, B. ovis, B. canis, and B. neotomae were purified and analyzed by SDS-PAGE, transmission electron microscopy and liquid chromatography coupled to mass spectrometry to determine the pan-proteome of these vesicles. In addition, antigenic proteins were detected by western blot with anti-Brucella sera. The in silico analysis of the pan-proteome revealed many homologous proteins, such as Omp16, Omp25, Omp31, SodC, Omp2a, and BhuA. Proteins contained in the vesicles from different Brucella species were detected by anti-Brucella sera. The occurrence of previously described immunogenic proteins derived from OMVs supports the use of these vesicles as candidates to be evaluated as an acellular brucellosis vaccine.
