Project description:Bone marrow-derived dendritic cells were infected with live or heat-killed Bordetella and cells were analyzed on day 4 post-infection cells were >90% CD11c-positive
Project description:Dendritic cells (DCs) are critical mediators of host defense against bacteria. The goal of this microarray study was to understand the global transcriptional response of bone marrow-derived dendritic cells (BMDCs) upon exposure to live bacteria, to better understand how DCs orchestrate a host-protective immune response. We found that BMDCs upregulate a number of critical immune-related genes upon exposure to live E. coli. Most notably, the gene encoding hepcidin, a critical regulator of mammalian iron homeostasis, was significantly upregulated in BMDCs upon exposure to live bacteria.
Project description:Mouse bone marrow derived dendritic cells were generated by culturing bone marrow cells at a density of 0.5x10E6 cells/ml in RPMI-1640 supplemented with 5% FCS, 1% Pen/Strep, 5microM 2-mercaptoethanol, 20ng/ml GM-CSF. At day 7 dendritic cells were stimulated or not with 500 ng/ml LPS, and collected at day 10.
2x10E8 cells were used to prepare whole cell extracts and to perform PU.1 immunoprecipitaion with PU.1 antibody (T-21 Santa Cruz). IgG was used as control.
Project description:Francisella tularensis is a highly infective Gram-negative bacterium, also known as the causative agent of tularemia. The disease is characterized by the delayed onset of the adaptive immune response which provides time for the bacterial replication. As an intracellular pathogen, Francisella proliferates mainly in the cytosol of host phagocytes, which are linkers between the innate and the adaptive immunity. In particular, dendritic cells (DCs) are known to be the most effective antigen-presenting cells and thus they are crucial for the induction of the adaptive response. However, Francisella is able to invade and to proliferate inside DCs while avoiding their effective activation and maturation. The goal of this study was to map phosphoproteome changes in DCs infected by Francisella in order to reveal host signaling pathways involved in Francisella-DC interaction. The focus was laid on early intervals (<1 h p.i.) because of the ability of Francisella to escape in this time period from the phagosome to the cytosol where it establishes its replicative niche. For comparison, attenuated and in vivo-protective Francisella strain was used in parallel in experiments. DC phosphoproteome was analyzed by the means of shotgun LC-MS-based proteomics and relative changes were quantified by the stable isotope labeling method specially developed for primary bone marrow-derived DCs.
Project description:To investigate dendritic cells-Leishmania interaction, the transcriptional profile of bone marrow-derived dendritic cells (BMDCs) infected with Leishmania infantum or of cells exposed to chemically inactivated parasites was assessed
Project description:The mechanisms by which dendritic cells (DCs) induce differentiation of naïve CD4+ T cells along the Th2 lineage is not well understood given that DCs themselves do not produce IL-4. In the present study, we undertook a microarray approach to identify genes involved in the induction of Th2 differentiation by DCs treated with a Th2-skewing adjuvant cholera toxin (CT). In the microarray analysis, murine bone marrow derived immature DCs were treated with CT. Of particular interest was tthe significant upregulation of the expression of c-kit. The upregulation of c-kit on DCs is critical for the induction of a Th2 response. Keywords: cholera toxin, bone marrow derived dendritic cells, gene expression array-based (RNA / spotted DNA/cDNA)