Project description:We evaluated the changes in the transcriptome of spleen and bone marrow isolated myeloid cells in healthy and melanoma bearing animals and melanoma cell lines, B16F10 and YUMM3.3 using RNA-sequencing technologies.
Project description:Transcriptional profiling of splenic lymphocytes derived from vaccinated mice, and ex-vivo exposed to M.tb.-infected macrophages in culture. Splenocytes from mice innoculated with Mycobacterium bovis BCG Pasteur (PAS), or M. bovis BCG Copenhagen (SSI), or heat killed BCG SSI (HK), or uninfected control, were ex vivo co-cultured with mouse bone marrow macrophages previously infected with M. tb.
Project description:The effect of TIM-3 stimulation was studied on B16F10 mouse melanoma cells, in vitro. Experiment Overall Design: Cy3: B16F10 mouse melanoma cells, treated by unspecific goat IgG (control) for 2 hours. Experiment Overall Design: Cy5: B16F10 mouse melanoma cells, in vitro, treated by an agonist goat polyclonal anti-mouse TIM3 antibody (RnD Systems) for 2 hours. Experiment Overall Design: Four biological replicates were used (in this study, biological replicate means different passage numbers of the same cell line). In each case, the treated sample (Cy5) was compared to its control one (Cy3) in Agilent dual-color microarray experiments.
Project description:Tumor-associated macrophages (TAMs) are key regulators in tumor progression. Although a role of bone marrow-derived monocytes (Mons) as TAM precursors has been revealed, the dynamic phenotypes of both TAMs and Mons regulated by the tumor microenvironment remain unclear. Here, we constructed an optimized micro-proteomics workflow, which was applicable to the mouse myeloid cells of low cell number. We sorted both TAMs and the corresponding Mons (1×105 per sample) from individual melanoma mouse model in the early stage and the late stage to establish the protein expression profiles by mass spectrum.
Project description:Bone marrow derived macrophages of Lysz-Cre;Catnbtm2Kem(fl/fl) mouse were compared with bone marrow derived macrophage of Catnbtm2Kem(fl/fl) control mouse Total RNA extracted from bone marrow derived macrophage
Project description:In vivo pathways of natural retinoid metabolism and elimination have not been well characterized in primary myeloid cells, even though retinoids and retinoid receptors have been strongly implicated in regulating myeloid maturation. Using a UAS-GFP reporter transgene, and retrovirally expressed Gal4-RARA in primary mouse bone marrow cells, we identified two distinct enzymatic pathways utilized by mouse myeloid cells ex vivo to synthesize RARA ligands from free vitamin A metabolites (retinyl acetate, retinol, and retinal). Bulk Kit+ bone marrow progenitor cells utilize diethylaminobenzaldehyde (DEAB)-sensitive enzymes, whereas bone marrow-derived macrophages use DEAB-insensitive enzymes to synthesize natural RARA-activating retinoids (all-trans retinoic acid). Bone marrow-derived macrophages do not express the DEAB-sensitive enzymes Aldh1a1, Aldh1a2, or Aldh1a3, but instead express Aldh3b1, which we found is capable of DEAB-insensitive synthesis of all trans-retinoic acid. However, under steady-state and stimulated conditions in vivo, diverse bone marrow cells and peritoneal macrophages showed no evidence of intracellular RARA-activating retinoids despite expression of these enzymes and a vitamin A sufficient diet, suggesting that the enzymatic conversion of retinal is not the rate limiting step in the synthesis of intracellular RARA-activating retinoids in myeloid bone marrow cells and that that RARA remains in an un-liganded configuration during adult hematopoiesis. In order to identify additional enzymes that might contribute to DEAB-insensitive retinoid synthesis in bone marrow-derived macrophages, we compared gene expression in Kit+ progenitor cells vs bone marrow macrophages by Affymetrix array profiling. We analyzed 3 biological samples each of mouse bone marrow-derived macrophages and Kit+ progenitor cells
