Mouse lung CD103+ and CD11b-high dendritic cell (DC) subsets
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ABSTRACT: Mouse lung CD11c+ dendritic cells are composed of 2 major DC subsets, the CD103+CD11b-low/intermediate DC (CD103+ DC) and the CD11b-highCD103- DC (CD11b-high DC). These 2 subsets are functionally distinct. Comparison of their functions showed CD103+ DC Microarray analysis was performed to compare the gene expression profiles of the 2 lung DC subsets in naïve mice.
Project description:Mouse lung CD11c+ dendritic cells are composed of 2 major DC subsets, the CD103+CD11b-low/intermediate DC (CD103+ DC) and the CD11b-highCD103- DC (CD11b-high DC). These 2 subsets are functionally distinct. Comparison of their functions showed CD103+ DC Microarray analysis was performed to compare the gene expression profiles of the 2 lung DC subsets in naïve mice. Perfused lungs from 6-8-week naïve BALB/cByJ mice were pooled and digested with collagenase D. CD11c+ cells were selected by anti-CD11c magnetic microbeads (Miltenyi) and stained by fluorochrome-conjugated mAb against I-A, CD103, CD11b, and CD11c plus 7-
Project description:Transcriptomic profiling of sheep skin lymph DC subsets after 16h in vitro culture in medium or stimulated by non replicative Canine Adenovirus serotype 2 Reference design: total enriched lymph dendritic cells- Biological replicates: 4 sheep (70958, 50274, 30066, 11261)- Samples: FACS sorted CD103 -type and CD11b type cultured in medium or with CAV2
Project description:Defense against attaching and effacing (A/E) bacteria requires the sequential generation of IL-23 and IL-22 to induce protective mucosal responses. While the critical source of IL-22 has been identified as CD4+ and Nkp46+ innate lymphoid cells (ILCs), the precise source of IL-23 is unclear. Here, we use genetic techniques to deplete specific classical dendritic cell (cDC) subsets and analyze immunity to the A/E pathogen Citrobacter rodentium. We find that Zbtb46+ cDCs, and specifically Notch2-dependent intestinal CD11b+ cDCs, but not Batf3-dependent CD103+ cDCs, are required for IL-23 production and immunity against C. rodentium. Notch2 controls cDC differentiation at a terminal step mediated by lymphotoxin signaling. Importantly, these results provide the first demonstration of a non-redundant function of CD11b+ cDCs in vivo. Analysis of Notch2-dependent genes in CD11b+ and DEC205+ splenic classical DC subsets. Splenocytes were harvested from littermate WT Notch2 f/f C57Bl/6 or Notch2 CD11c-cre C57Bl/6 mice and DC subsets sorted to >95% purity on the FACSAriaII.
Project description:We hypothesize that under homeostatic as well as inflammatory conditions circulating monocytes and/or their bone marrow-derived progenitors might contribute to the replenishment of CD103+ and CD103- DC in lymphoid and non-lymphoid compartments. To that end, bone marrow cells from CX3CR1+/gfp C57BL/6 mice were sorted as follows: lineage negative (CD3, CD19, NK1.1, Ter119, Ly6G and CD11c) CX3CR1+c-kit+. Sorted cells were further cultured in vitro under the continuous presence of GM-CSF. lin-CX3CR1+c-kit+ bone marrow cells gave rise to CD103+ and CD103- DC in vitro. To test whether CD103 might be a suitable marker that allows the differentiation of functionally distinct DC subsets generated in vitro, CD11c+CD103+ and CD11c+CD103- DC were sorted from day 5 cultures of lin-CX3CR1+c-kit+ cells and analyzed by whole mouse genome microarrays from Agilent technologies. Compared to CD103+ DC, CD103- DC displayed a strong up-regulation of transcripts for genes involved in innate immunity, whereas those involved in costimulation were down-modulated. Our data suggest distinct functional activity of these two DC subsets. Keywords: Transcriptional profiling-Cell type comparison We compared the transcriptional profile of CD11c+CD103+ and CD11c+CD103- dendritic cells by use of Agilent Whole Mouse Genome Microarrays. Two sets of samples were generated independently as real biological replicates. The microarray analysis was performed as a dual-color experiment, including a dye-swap.
Project description:We hypothesize that under homeostatic as well as inflammatory conditions circulating monocytes and/or their bone marrow-derived progenitors might contribute to the replenishment of CD103+ and CD103- DC in lymphoid and non-lymphoid compartments. To that end, bone marrow cells from CX3CR1+/gfp C57BL/6 mice were sorted as follows: lineage negative (CD3, CD19, NK1.1, Ter119, Ly6G and CD11c) CX3CR1+c-kit+. Sorted cells were further cultured in vitro under the continuous presence of GM-CSF. lin-CX3CR1+c-kit+ bone marrow cells gave rise to CD103+ and CD103- DC in vitro. To test whether CD103 might be a suitable marker that allows the differentiation of functionally distinct DC subsets generated in vitro, CD11c+CD103+ and CD11c+CD103- DC were sorted from day 5 cultures of lin-CX3CR1+c-kit+ cells and analyzed by whole mouse genome microarrays from Agilent technologies. Compared to CD103+ DC, CD103- DC displayed a strong up-regulation of transcripts for genes involved in innate immunity, whereas those involved in costimulation were down-modulated. Our data suggest distinct functional activity of these two DC subsets. Keywords: Transcriptional profiling-Cell type comparison
Project description:RNA-seq analyses of two intestinal dendritic cell subsets, CD103+CD11b- and CD103-CD11b+, from wild type and CbfbF/F:CD11c-Cre mice.
Project description:We report phenotypes in gut dendritic cell development by loss of Runx/Cbfb transcription factor complexes. To further examine function of residual CD103+CD11b- and CD103-CD11b+ DCs, we performed RNA-seq analyses and compared gene expression signatures between control and Cbfb-deficient cells.
Project description:Background: Dendritic cells (DCs) are critical for regulating CD4 and CD8 T cell immunity, controlling Th1, Th2, and Th17 bias, generating inducible Tregs, and inducing tolerance. Multiple DC subsets have been identified in the mouse that are thought to have evolved to control these different immune outcomes. However, how these subsets differentially respond to inflammatory and/or tolerogenic signals in order to accomplish their divergent functionality remains unclear. Results: We analysed the responses of murine, splenic CD8 and CD11b DC subsets to in-vivo stimulation with lipopolysaccharide using RNA-Seq and systems biology approaches and observed responses are highly subset-specific. We reanalysed multiple datasets from the literature and show that these subset responses are obscured when analysing signaling at the population level. We show that the subset-specificity is due to the unique regulation of distinct TLR4 pathway modulators that ‘fine-tune’ a common TLR4 cascade rather and not due to major differences in signaling pathways or transcription factors. Conclusions: We propose the Pathway Modulation Model wherein common signaling pathways are regulated by unique sets of modulators allowing for distinct immune responses in closely related DC subsets. We extend these observations using analagous datasets from the literature and show that our model provides a global mechanism for generating cell subset-specific signaling in multiple subpopulations in mouse and man. Splenic CD8 and CD11b DC subsets from LPS stimulated (10 pooled animals) and Control (5 pooled animals) mice were analysed by RNA-Seq.
Project description:To investigate whether differentially expressed genes induced by SIIN-Q11 were shared between lung CD11b+ and CD103+ DCs, we performed gene expression profiling analysis of lung CD11b+ and CD103+ DCs after intranasal SIIN-Q11.
Project description:Transcriptome analysis of dentritic cells from the spleen (CD11c+) or MLN (CD11c+CD103+) of mice with DC-specific deletion of TGFBR2 gene, or their control littermates. TRGFR2 gene was deleted in dendritic cells using Cre/lox approach. Mice with this deletion develop spontaneous multi-organ autoimmune inflammation and die by 15 weeks of age. Splenic CD11c+ Dc were isolated by magetic cell sorting. MLN CD11c+CD103+ DC were flow sorted. RNA was isolated using RNAqueous-Micro kit (Ambion) and analyzed using Affymetrix Mouse Exon 1.0 ST Array