Project description:TDC are an hematopoietic cell subset identified and characterized by Mirela Kuka, Ivana Munitic and Jonathan D. Ashwell (Kuka et al., 2012, Nat Commun). TDC combine dendritic cell (DC) and conventional ab T cell markers and functional properties. Such duality might render TDC particularly responsive to infectious organisms, because they can bridge innate and adaptive traits. This novel cell subset substantially differs from other known innate T cells in many ways: they undergo an antigen-specific selection process indistinguishable from classical ab cells, they can proliferate in response to cognate antigens, and they lack markers of NKT or gd T cells. The expression data found in this dataset confirm that TDC have a genetic signature distinct from that of classical DC or of conventional T cells. This distinct genetic profile identifies them as a separate cell type and raises the possibility that TDC may have functions other than those of T and DC.
Project description:TDC are hematopoietic cells that combine dendritic cell (DC) and conventional T cell markers and functional properties. To investigate the gene expression profile of TDC, we performed a bulk RNA sequencing (RNAseq) on TDC from Zbtb46-GFP reporter mice.
Project description:Inflammatory cytokines promote the accumulation of activated CD8 T cells. Here, we transfer 600 OT-I CD8 T cells iv into naïve C57BL/6 hosts. One day later, 500,000 LPS-matured and OVA257 peptide-coated DC were injected iv into OT-I CD8 T cell seeded hosts with (DC+CpG) or without (DC). Other seeded mice were infected with 2x10^4 virulent Listeria monocytogenes (vLM-OVA) iv. OT-I CD8 T cells were harvested from the spleen, flow sort purified, then RNA was extracted using RNeasy (Qiagen) kit. Naive OT-I CD8 T cells (Naive) were purified from the spleens of OT-I transgenic mice. Each group had three independent biological replicates.Transcriptomes were compared using DAVID analysis (with genes scoring FDR<0.01) and GSEA analysis.
Project description:Inflammatory cytokines promote the accumulation of activated CD8 T cells. Here, we transfer 600 OT-I CD8 T cells iv into naïve C57BL/6 hosts. One day later, 500,000 LPS-matured and OVA257 peptide-coated DC were injected iv into OT-I CD8 T cell seeded hosts with (DC+CpG) or without (DC). Other seeded mice were infected with 2x10^4 virulent Listeria monocytogenes (vLM-OVA) iv. OT-I CD8 T cells were harvested from the spleen, flow sort purified, then RNA was extracted using RNeasy (Qiagen) kit. Naive OT-I CD8 T cells (Naive) were purified from the spleens of OT-I transgenic mice. Each group had three independent biological replicates.Transcriptomes were compared using DAVID analysis (with genes scoring FDR<0.01) and GSEA analysis. 3 biological replicates per group. Groups included Naïve OT-I CD8 T cells, DC+CpG OT-I CD8 T cells, DC OT-I CD8 T cells, and vLM-OVA OT-I CD8 T cells. Most comparisons used Naïve OT-I CD8 T cells as a baseline comparison
Project description:To evaluate gene expression profiles on different dendritic cell subsets isolated from spleens of mice We used whole genome microarrays to identify genes specific to pDCs and not other DC subsets in the absence of disease pDCs, CD11b+ cDCs, and CD8+ cDCs were FACS-purified from spleens of healthy C57bl/6 mice
Project description:The aim was to study the transcriptional profiling of the tdc cluster delection mutant E. faecalis V583 Î?tdc (non-tyramine producer) compared to the wild type strain E. faecalis V583 (tyramine producer). We compared the expression profile of the strains grown in M17 medium with glucose as carbon source and suplemented with tyrosine. E. faecalis V583 Î?tdc cells (test) compared with E. faecalis V583 cells (reference). Both strains grown in GM17 medium suplemented 15 mM tyrosine.
Project description:High-dimensional approaches revealed emerging heterogeneity within dendritic cells (DC), including a population of transitional DC (tDC) present in mouse and human. However, tDC origin and relationship to other DC subsets are not fully understood. Here, we show that tDC are distinct from other well-characterized DC and conventional DC precursors (pre-cDC). We demonstrate that tDC originate from bone marrow progenitors shared with plasmacytoid DC (pDC). In the periphery, tDC contribute to the pool of ESAM+ type 2 DC (DC2), and these DC2 harbor pDC-related developmental features. Different from pre-cDC, tDC have lower turnover, capture antigen, respond to stimuli, and activate antigen-specific naïve T cells, all characteristics of differentiated DC. Different from pDC, viral sensing by tDC results in IL-1b secretion and fatal immune pathology in a murine coronavirus model. Our findings suggest that tDC are a distinct pDC-related subset with a DC2 differentiation potential and unique pro-inflammatory function during viral infections.
Project description:Plasmacytoid dendritic cells (pDC) are the main source of type I interferon (IFN) during viral infections. Their other functions are debated, due to a lack of tools to identify and target them in vivo without affecting pDC-like cells and transitional DC (tDC), which harbor overlapping phenotypes and transcriptomes but a higher efficacy for T cell activation. To overcome this bottleneck, we designed, generated and validated a pDC-Tomato reporter mouse. We bred pDC-Tomato with Zbtb46 GFP mice to yield the he ZeST mouse strain that enabled transcriptomic profiling of all splenic DC types, by single cell RNA sequencing, both at steady state and during the course of the infection with mouse cytomegalovirus (MCMV). Analyses of the transcriptomic dataset unraveled diverging activation of pDC-like cells vs tDC during the infection. This dataset and the associated specific gene modules will be useful to delineate the physiological functions of pDC versus other DC types.
Project description:High-dimensional approaches have revealed emerging heterogeneity within dendritic cells (DC), including a population of transitional DC (tDC) present in mouse and human. However, tDC origin and relationship to other DC subsets are not fully understood because their phenotype partially overlaps with previous definitions of conventional DC precursors (pre-cDC). Here, we show that tDC are distinct from other well-characterized DC and pre-cDC. By combining single-cell transcriptomics, high-dimensional immunophenotyping, lineage tracing and adoptive transfer experiments, we demonstrate that murine tDC originate from bone marrow progenitors shared with plasmacytoid DC (pDC) that are distinct from CD115-expressing conventional DC progenitors. In the periphery, tDC contribute to the pool of Esam+ type 2 DC (DC2), and these tDC-derived DC2 harbor pDC-related developmental features. tDC have lower turnover than pre-cDC2, capture antigen, respond to stimuli, and potently antigen-specific naïve T cells, all characteristics of differentiated DC. Different from pDC, viral sensing by tDC results in IL-1β secretion and fatal immune pathology in a model of murine coronavirus. Our findings suggest that tDC are a distinct pDC-related lineage with a DC2 differentiation potential and unique pro-inflammatory function during viral infections. The objective of this experiment was to evaulate the transcriptional changes in various dendritic cell subpopulations
Project description:To evaluate gene expression profiles on different dendritic cell subsets isolated from spleens of mice We used whole genome microarrays to identify genes specific to pDCs and not other DC subsets in the absence of disease