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:Brucella abortus (B. abortus), an intracellular bacterium, is the causative agent of Brucellosis. This organism invades into macrophages and then survives through its abilities to modulate host cells functions. The biggest problem caused by B. abortus is that it prevents macrophage elimination and makes it difficult to remove B. abortus from the host body. Therefore, it is essential to identify the bacterial genes involved in virulence factor as a first step to understanding the bacterial pathogenicity and controlling Brucellosis. To identify these genes, B. abortus mutant strains were generated using transposon mutagenesis and transcriptomic profile during macrophage infection were analyzed. The gene expression level was analyzed using total RNA obtained from THP-1 cells infected with B. abortus wild type and mutant strains and cellular immunity during the infections were compared to wild type infected cell to identify the role of genes in B. abortus pathogenicity. Transcriptomic profiling showed that two mutant strains having disrupted genes related to 4-hydrobenzoate 3-monooxygenase (PHBH) of C1 strain and heme exporter protein cytochrome C (CcmC) of C10 strain, induced suppression of cytokine expression during infection in human macrophages. Conversely, two other mutant strains of exopolyphosphatase (PPX)of C27 and Peptidase M24 of C32 induced activation of cytokine expression in the THP-1 macrophage cells.
