RNA-Seq of Group 2 innate lymphoid cells (ILC2) from lung and lymph node
Ontology highlight
ABSTRACT: Group 2 innate lymphoid cells (ILC2) promote the production of a type-2 immunological environment in the uterus and have tissue-specific gene signatures that change with pregnancy.
Project description:Group 2 innate lymphoid cells (ILC2) promote the production of a type-2 immunological environment in the uterus and have tissue-specific gene signatures that change with pregnancy.
Project description:Oral tolerance prevents pathological inflammatory responses towards innocuous foreign antigens via peripheral regulatory T cells (pTreg cells). However, whether a particular subset of antigen-presenting cells (APCs) is required during dietary antigen exposure to instruct naïve CD4+ T cells to differentiate into pTreg cells has not been defined. Using myeloid lineage-specific APC depletion in mice, we found that monocyte-derived APCs are dispensable, while classical dendritic cells (cDCs) are critical for pTreg cell induction and oral tolerance. CD11b¬â?? cDCs from the gut-draining lymph nodes efficiently induced pTreg cells, and conversely, loss of IRF8-dependent CD11bâ?? cDCs impaired their polarization, although oral tolerance remained intact. These data reveal the hierarchy of cDC subsets in pTreg cell induction and their redundancy during oral tolerance development. Sorted naïve CD45.1 OT-II CD4 T cells were co-cultured with four dendritic cell subpopulations sorted from mouse mesenteric lymphnodes. 24h later OT-II cells were sorted again and compared in their gene expression profile.
Project description:IL-2 signals into CD8 T cells have a programming and regulatory role in driving cells to full effector and memory differentiation. This study was designed to look for IL-2 target genes that affect CD8 T cell responses. Experiment Overall Design: Treatment of mice with IL-2/anti-IL-2 immune complexes (IL-2 complex) stimulates all fraction of CD8 T cells in vivo. Because STAT5 phosphorylation peaked 1 h after injection of the IL-2 complex, gene expression in CD8 T cells was compared at 0 h (no treatment), 1 h and 3 h post treatment. Two mice were used for each time point.
Project description:Innate lymphoid cells (ILCs) play critical roles during innate immune responses to pathogens and lymphoid organ development. IL-7Ra+ ILC subsets, similar to T helper (Th) cell subsets, produce distinctive effector cytokines. The molecular control of IL-7Ra+ ILC development and maintenance has yet to be dissected. Here we report that GATA3 is indispensable for the development of all IL-7Ra+ ILC subsets and T cells. Gata3 conditional deficient mice have no lymph nodes and are susceptible to Citrobactor rodentium infection. Genome-wide gene analyses indicate that GATA3 regulates similar set of cytokines and receptors in ILC2s and Th2 cells and is critical for the maintenance of ILC2s. Thus, GATA3 plays parallel roles in establishing and regulating both adaptive and innate lymphocytes. To identify GATA3 regulated genes in type 2 innate lymphoid cells by tamoxifen-mediated acute deletion of Gata3 gene.
Project description:Gene-expression microarray datasets generated as part of the Immunological Genome Project (ImmGen). Primary cells from multiple immune lineages are isolated ex-vivo, primarily from young adult B6 male mice, and double-sorted to >99% purity. RNA is extracted from cells in a centralized manner, amplified and hybridized to Affymetrix 1.0 ST MuGene arrays. Protocols are rigorously standardized for all sorting and RNA preparation. Data is released monthly in batches of cell populations. This Series record provides access to Immunological Genome Project data submitted to GEO.
Project description:To identify possible transcriptional changes induced by NK cell education in this reductionistic model, we performed single cell RNA sequencing (scRNA-seq) on single positive Ly49A (NKG2A-Ly49G2-Ly49I-Ly49C-, denoted as sp-Ly49A, NK cells from Dd-/-, Dd+/- and Dd+/+ mice. To ensure minimum batch-effects, sorted sp-Ly49A cells from Dd-/-, Dd+/-, and Dd+/+ mice were hash-tagged with oligo-labelled CD45 antibodies, pooled and sequenced together.
Project description:MPP2, MPP3 and CLP populations were isolated by fluorescence-activated cell sorting (FACS) from the BM of B6 mice based on the surface expression of VCAM1, FLT3 and IL7Ra within LSK population. On average 100K cells were analyzed for each population in 3 independent biological experiments. We generated sequence libraries from these low RNA inputs using the SMARTer Stranded RNA-Seq and sequenced at high depth of coverage to generate ~2x120 M reads per biological replicate.
Project description:During embryogenesis, development of hematopoietic stem cells (HSC) occurs in the fetal liver and involves coordinate programs of transcription. Taspase1, a highly conserved threonine protease, directly cleaves and regulates the TFIIA families of transcription factors. We discovered that loss of Taspase1 (Tasp1-/-) or non-cleavage of TFIIAa-b (TFIIAa-b nc/nc) leads to a severe fetal liver developmental retardation that is associated with impaired HSC self-renewal and loss of HSC quiescence. We used microarray to elucidate the mechanism(s) by which TFIIA regulates fetal liver hematopoiesis, and expression of targets of HoxA9 was found to be altered by gene set enrichment analyses. Embryonic day 14.5 fetal liver HSCs (defined as Lineage-Sca-1+c-Kit+CD150+cells) of wild-type (n = 4) and TFIIAa-b nc/nc (n = 3) were analyzed.