Project description:Human blood CD1c+ cDC2s can be divided into two functionally distinct subpopulations according to their CD5 expression. CD5high and CD5low cDC2s differ significantly in their gene expression, cytokine production, antigen presentation, and T cell polarization. However, the plasticity of CD5high and CD5low cDC2s was unknown. We found that about 40% CD5low cDC2s upregulated their CD5 expression when cultured in the presence of TGF-β. Transcriptome analysis revealed that the converted CD5high cDC2s are closer to cultured CD5high cDC2s, separating form unconverted CD5low cDC2s. Converted CD5high cDC2s with higher IRF4 upregulated some steady-state DC mature molecules, such CCR7, CD86, PD-L1 and CCL22. Unconverted CD5low cDC2s with higher MAFB upregulated some monocyte signature genes, such CCR2, CSF1R and CCL2. Moreover, Converted CD5high and unconverted CD5low cDC2s were significantly different in inducing CD4+ T cell polarization. Specifically, converted CD5high cDC2s recruited and induced more Treg cells than unconverted CD5low cDC2s. We also found that the ratio of CD5high cDC2s are significantly higher in colorectal tumor tissue comparing with paired paratumor tissue. In summary, TGF-β can effectively promote the conversion from CD5low cDC2s to CD5high cDC2s accompanied by phenotype and function change. This may be another mechanism which contributes to TGF-β induced immune suppression.
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:The human dendritic cell (DC) family has recently been expanded by CD1c+CD14+CD163+ DCs, introduced as DC3. DC3 are found in tumors and peripheral blood of cancer patients but which cells can serve as their pre-cursors remain unknown. CD1c+CD14+ cells share similarities with both CD1c+ DCs (cDC2s) and CD14+ monocytes on transcriptomic and phenotypic level. To investigate whether CD14+ cDC2s are closer related to CD14- cDC2s or monocytes on transcriptomic level, we analyzed their RNA.
Project description:We analyzed the transcriptome differences of wild-type, CD97- and Gα13-deficient (Adgre5-/- and CD11c-cre x Gna13fl/fl) type-2 conventional dendritic cells (cDC2s) in spleen. Three technical repeats of ~10^5 cells per sample from each of one mouse were included. Compared to wild-type cDC2s, CD97- and Gα13-deficient cDC2s differentially expressed many genes, but CD97- and Gα13-deficient cDC2s were almost the same. GSEA showed that Mrtf-a dependent genes were upregulated in CD97- and Gα13-deficient cDC2s, which code for cytoskeleton proteins. These data support that CD97-Gα13 signaling regulates splenic cDC2 motility by the actin cytoskeleton.
Project description:The goal of this study was to determine whether there are any gene expression changes in cDC1s and cDC2s from WT, Flt3 KO, or Flt3L KO mice. Specifically whether developing in the absence of Flt3 signaling had any effects on the gene expression of the cDCs
Project description:To investigate the role of Notch2 in the inflammatory response in chondrocytes, we cultured primary chondrocytes derived from wild type and Notch2 mutant mice and treated TNFalpha or vehicle. We then performed gene expression profiling analysis using data obtained from RNA-seq of wild type and Notch2 mutant chondrocytes.
Project description:Primary Sjögren’s syndrome (pSS) is a systemic autoimmune disease characterized by lymphocytic infiltration of the exocrine glands. Conventional dendritic cells (cDCs)-2 play a critical role in T and B cell activation, leading to germinal centre formation and autoantibody production. To understand the mechanisms underlying cDC2 dysregulation in pSS we performed RNA-sequencing analysis and functional validation on circulating cDC2s from pSS, non-Sjögren’s sicca (nSS) patients and healthy controls (HC). Two independent cohorts were established to identify reproducible dysregulated signatures, which included the interferon (IFN), toll-like receptors (TLR) signaling and antigen processing and presentation pathways. We confirm by flow cytometry that pSS-cDC2s were less efficient to degrade BSA but more efficient to uptake BSA both linked with the presence of anti-SSA antibodies. As the majority of the SSA+ pSS patients exhibit an IFN-signature, we tested whether IFNα priming would influence cDC2s antigen-uptake and processing. IFNα priming increased cDC2s uptake capacity, but not antigen processing. Additionally, pSS-cDC2s showed an increased uptake capacity of apoptotic salivary gland epithelial cells. Finally, pSS-cDC2s increased the proliferation of HC CD4+ T cells and the expression of CXCR3 and CXCR5 on the proliferating HC CD4+ T cells, contributing to T cell migration into the inflamed salivary glands. Here we provide the first in-depth molecular characterization of pSS-cDC2s and show that the transcriptomic and functional alterations observed in pSS-cDC2s are linked to IFN-signature. In view of its role in pSS immunopathology, delineating the molecular networks that drive cDC2s holds promise for targeting these cells in pSS.