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Dectin1-Stimulated mTOR-C/EBPβ Signaling Induces IL-10-Producing Regulatory Tr2 Cells

ABSTRACT: In the present study we describe a new subset of murine T cells producing high levels of IL-10 that, in this case, are induced by C-type lectin receptor-stimulated (i.e., Dectin-1-stimulated) dendritic cells. This T cell produces IL-10 as a result of a transcriptional mechanism that is distinct from that inducing IL-10 in Th2 cells, Tr1 cells or other T cell subsets. In particular, the mechanism involves mTOR signaling and induction of the LIP isoform of C/EBPb, a factor that interacts with phospho-CREB and GATA3 to induce transactivation of the IL-10 gene. These T cells have strong regulatory properties in relation to fungal infection and hence have been termed Tr2 regulatory T cells to distinguish them from other regulatory T cell subsets.

ORGANISM(S): Mus musculus

PROVIDER: GSE87673 | GEO | 2017/10/14

SECONDARY ACCESSION(S): PRJNA345513

REPOSITORIES: GEO

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Mus musculus

Project description:Dectin1-Stimulated mTOR-C/EBPβ Signaling Induces IL-10-Producing Regulatory Tr2 Cells

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Project description:C/EBPb is an auto-repressed protein that becomes posttranslationally activated by Ras-MEK-ERK signalling. C/EBPb is required for oncogene-induced senescence (OIS) of primary fibroblasts, but also displays pro-oncogenic functions in many tumour cells. Here, we show that C/EBPb activation by H-RasV12 is suppressed in immortalized/transformed cells, but not in primary cells, by its 30 untranslated region (30UTR). 30UTR sequences inhibited Ras-induced cytostatic activity of C/EBPb, DNA binding, transactivation, phosphorylation, and homodimerization, without significantly affecting protein expression. The 30UTR suppressed induction of senescence-associated C/EBPb target genes, while promoting expression of genes linked to cancers and TGFb signalling. An AU-rich element (ARE) and its cognate RNA-binding protein, HuR, were required for 30UTR inhibition. These components also excluded the Cebpb mRNA from a perinuclear cytoplasmic region that contains activated ERK1/2, indicating that the site of C/EBPb translation controls de-repression by Ras signalling. Notably, 30UTR inhibition and Cebpb mRNA compartmentalization were absent in primary fibroblasts, allowing Ras-induced C/EBPb activation and OIS to proceed. Our findings reveal a novel mechanism whereby non-coding mRNA sequences selectively regulate C/EBPb activity and suppress its anti-oncogenic functions. NIH-3T3 cells were retrovirally infected and selected with appropriate antiobiotics. Equal number of cells were plated and collected 48-72 hours later.

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Influence of C/EBPβ 3'UTR on RasV12 induced gene expression

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ATAC-seq examining chromatin accessibility of mouse splenic IL-10+ and IL-10-CD19+CD21hiCD24hi B cells

Project description:Chromatin profiling of mouse splenic ex-vivo IL-10+, IL-10-CD19+CD21hiCD23hiCD24hi B cells and IL-10- follicular B cells, after antigen induced arthritis. IL-10eGFP reporter (Vert-X) mice were used to sort IL-10+ and IL-10- subsets. ATAC-seq was used in conjunction with microarray data to understand the chromatin profiles and transcriptional landscape of mouse regulatory B cells, compared to other B cell subsets.

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Project description:Type 1 regulatory T (Tr1) cells are one of the regulatory T cell subsets that are characterized by the production of high amount of IL-10 and lack of FOXP3 expression. Lymphocyte-activation gene 3 (LAG3) is a CD4 homologue molecule and we have previously reported that LAG3 is expressed on IL-10 producing regulatory T cells. However, naturally occurring Tr1 cells in human secondary lymphoid tissue have not been detected. We identified CD4+CD25-LAG3+ T cells in human tonsil. We compared mRNA expression of five CD4+ T cell subsets in tonsil using microarray analysis (CD4+CD25-LAG3+ T cells, CD4+CD25-CXCR5+PD-1+ follicular helper T cells (TFH), CD4+CD25+ T cells, CD4+CD25-LAG3-CD45RO+ cells and CD4+CD25-LAG3-CD45RO- cells). A human tonsil was obtained from a patient undergoing routine tonsillectomy, and five tonsillar CD4+ T cell subsets were sorted (each 1 x 10^5 cells). There is no biological replication.

2016-04-01 | E-GEOD-73927 | biostudies-arrayexpress

Proteomic profiling of subsets of human primary NK cell-derived extracellular vesicles by LC-MS/MS

Project description:In this study we have compared the proteomic profile of subsets of extracellular vesicles (EVs) prepared from primary human NK cells cultured for 48hrs in serum-free conditions supplemented with 10 ng/ml human recombinant IL-12, IL-15, and IL-18. The aim was to isolate and define an EV subset with cytolytic activity against tumor cells. For this purpose, bulk EVs were separated according to density (density gradient ultracentrifugation; DG-UC) to yield 3 distinct subsets.

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