Project description:We deciphered the transcriptional landscape of Peyer's patch CD11c+ phagocytes from their emergence in the tissue to their final maturation state at homeostasis and under stimulation with the TLR7 ligand R848 and the TLR9 ligand CpG.
Project description:The initiation of the mucosal immune response in Peyer’s patch (PP) relies on the sampling, processing and efficient presentation of foreign antigens by dendritic cells (DC). PP DC encompass five subsets, among which CD11b+ conventional DC (cDC) and LysoDC have distinct progenitors and functions but share many cell surface markers. This has previously led to confusion between these two subsets. In addition, another PP DC subset, termed double-negative (DN), remains poorly characterized. Here, we have studied the genetic relatedness of the different subsets of PP cDC at steady state and under TLR7 ligand stimulation. We also provide the transcriptional profiles of subepithelial TIM-4- and interfollicular TIM-4+ macrophages.
Project description:Gene expression data of Peyer's patch LysoDC differentiation states and macrophages at steady state and under TLR7 ligand stimulation
Project description:PeyerM-bM-^@M-^Ys patches (PP) are primary inductive sites of mucosal immunity. Defining PP mononuclear phagocyte system (MPS) is thus crucial to understand the initiation of mucosal immune response. We provide a comprehensive analysis of the phenotype, distribution, ontogeny, lifespan, function and transcriptional profile of PP MPS. We show that monocytes give rise to macrophages and to lysozyme-expressing DC (LysoDC) which are both involved in particulate antigen uptake, display strong innate antiviral and antibacterial gene signatures and, upon TLR7 stimulation, secrete IL-6 and TNF but no IL-10. However, unlike macrophages, LysoDC display a rapid renewal rate, strongly express genes of the MHCII presentation pathway and prime naM-CM-/ve helper T cells for IFNg production. Our results show that in PP, at steady state, monocytes generate both LysoDC and macrophages which display distinct features from their adjacent villus counterparts. 3 replicates of 3 different mononuclear phagocyte subsets have been extracted from Peyer's Patches of WT C57Bl/6 mice: CD11b+ DC, lysoDC and lysoMac. The total RNA of PP-sorted cells from the 3 independent experiments was extracted with a Qiagen micro RNAeasy PLUS kit. Quantity, quality and absence of genomic DNA contamination were assessed with a Bioanalyser (Agilent). Microarray experiments were performed by the Plateforme Biopuces of Strasbourg (France) using the GeneChipM-BM-. Mouse Gene 1.0 ST array.
Project description:To investigate how stimulation from indigenous bacteria for a day affects the local transcriptome of both Peyer's patch and surrounding ordinary mucosa in the rat ileum, we treated rats with a nonabsorbable antibiotics cocktail (vancomycin, polymyxin B and sucrose) for antibiotics-treatment group or sucrose solution for control group in drinking water 1 day before euthanasia. We then performed gene expression profiling analysis using data obtained from RNA-seq of ileal Peyer's patch and surrounding ordinary tissue from rats with or without antibiotics treatment.
Project description:Previous studies have shown that purified dendritic cells (DCs) have cell-intrinsic, age-dependent differences in their response to TLR stimulation. To delineate which aspects of the age-dependent difference in innate immunity are cell intrinsic vs extrinsic, we searched for global differences to TLR7/8 stimulation in purified adult vs neonatal DC populations. We hypothesize that very few selected cell intrinsic differences in gene expression of key immune genes between these 2 age groups exist, and the bulk would be cell extrinsic differences. The results show that there are age-dependent differences in expression of several key genes involved in the immune response at baseline already. Upon stimulation, we identified a substantially larger fraction of age-dependent differentially expressed genes in conventional than plasmacytoid DCs. Bioinformatics analyses indicate that important immune pathways were significantly differentially expressed in DC subsets between the 2 age groups. Total RNA was isolated from purified human conventional and plasmacytoid dendritic cells from 6 adult and 6 cord blood donors that were stimulated with 3M-003 at a final concentration of 5 uM for 1 and 6 hr.
Project description:The experiments were designed to gain insight into the gene expression of conventional and unconventional murine T cell subsets in the steady state and upon activation TCRαβ and TCRγδ intraepithelial lymphocytes (IEL) isolated from murine gut, naïve and virus-induced memory splenocytes, activated dendritic epidermal T cells (DETC)
Project description:Monocytes and macrophages are essential cells in the early response in their capacity as ubiquitous phagocytic cells. They phagocytose microorganisms or damaged cells and sense pathogen/damage-associated molecular patterns (PAMPs/DAMPs) through innate receptors such as Toll-like receptors (TLRs). We investigated a phenomenon where co-signaling from TLR2 and TLR8 in human primary monocytes provides a distinct immune activation profile compared to signaling from either TLR alone. Comparison of gene signatures induced by either stimulus alone or together, show that co-signaling result in downstream differences in regulation of signaling and gene transcription. To investigate the interaction of TLR2 and TLR8 stimulation primary human monocytes isolated from buffy coat were stimulated with TLR2/6 ligand FSL-1 and/or TLR7/8 ligand CL075 and the gene expression monitored at 1,2 and 3h post stimulation.
