TGF-β promotes the conversion of human CD5low cDC2s to tolerogenic CD5high cDC2s
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ABSTRACT: 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: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:We analyzed the transcriptome differences of wild-type and IRF4-deficient (CD11c-cre x Irf4fl/fl) type-2 conventional dendritic cells (cDC2s) in spleen. Wild-type or IRF4-deficient (CD11c-cre x Irf4fl/fl) bone marrow cells were transferred into lethal irradiated CD45.1 mice. 8 weeks later, splenic cDC2s were purified and prepared for RNA-seq analysis. Three technical repeats of ~10^5 cells per sample from each of one mouse were included. CD97 was downregulated in IRF4-deficient cDC2s. GSEA showed that upregulated and downregulated genes in CD97-deficient cells were strongly enriched in IRF4-regulated genes. These data are in accord with CD97 acting within the IRF4 gene-expression network.
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 and ArhGEF1-deficient (Arhgef1-/-) type-2 conventional dendritic cells (cDC2s) in spleen. Wild-type or ArhGEF1-deficient (Arhgef1-/-) bone marrow cells were transferred into lethal irradiated CD45.1 mice. 8 weeks later, splenic cDC2s were purified and prepared for RNA-seq analysis. The gene expression profiles of CD97-, Gα13- and ArhGEF1-deficient cDC2s were highly similar, providing evidence that these molecules function in the same pathway.
Project description:There are three major dendritic cell (DC) subsets in both human and mouse, plasmacytoid DCs (pDCs) and two types of conventional DCs (cDCs), cDC1s and cDC2s. cDC2s are important for polarizing CD4+ naive T cells into different subsets including Th1, Th2, Th17, Th22 and regulatory T cells (Tregs). In mice, cDC2s can be further divided into phenotypically and functionally distinct subgroups. However, subsets of human cDC2s have not been reported. In the present study, we showed that human blood CD1c+ conventional DCs (cDC2s) can be further separated into two subpopulations according to their CD5 expression status. Comparative transcriptome analyses showed that the CD5high DCs expressed higher levels of cDC2-specific genes, including IRF4, which is essential for the cDC2 development and its migration to lymph nodes. In contrast, CD5low DCs preferentially expressed monocyte-related genes, including the lineage-specific transcription factor MAFB. Furthermore, compared with CD5low subpopulation, CD5high subpopulation showed stronger migration toward CCL21 and overrepresentation among migratory DCs in lymph nodes. Additionally, the CD5high DCs induced naïve T-cell proliferation more potently than the CD5low DCs. Moreover, CD5high DCs induced higher levels of IL-10-, IL-22- and IL-4-producing T-cell formation, whereas CD5low DCs induced higher levels of IFN-γ-producing T-cell formation. Thus, we show that human blood CD1c+ cDC2s encompass two subsets that differ significantly in phenotype, gene expression, and functions. We propose that these two subsets of human cDC2s could potentially play contrasting roles in immunity or tolerance.
Project description:DNA from mature sperm was obtained from a mouse heterozygous for a mutation in Setdb1 (MommeD13) and a wildtype control mouse. The DNA was bisulfite converted and sequenced on a HiSeq 2000 to ~30x coverage. After mapping the resulting reads to the mouse genome DNA methyalation values at CpG dinucleotides were obtained as the ratio of reads with unconverted Cs relative to the sum of converted and unconverted reads. The resulting values were used to identify sites with a shift in the percentage of methylated CpGs between wildtype and mutant sperm.
Project description:Conventionaldendritic cells (cDCs) in the lung that drive effector T cell differentiation, and initiate allergic responses upon inhalation of allergens. CD11b+type 2 cDCs (cDC2s) are heterogeneous, but the relationship among subpopulations has been elusive. To analyze their developmental pathways of cDC2 subpopulations, we performed single cell RNA-sequencing (scRNAS-Seq) of total lung cDC2s at various times following inhalation of house dust extracts, and detected multiple clusters based on their differential transcriptomes. The data of cDC2 transcriptome at steady state, early and later stages after activation will be applicable for the maturation pathway analysis of cDC2s.