Treg cell gene expression signature in B16 melanoma
ABSTRACT: Gene expression signature of Treg cells in B16 melanoma was measured and compared to B16-infiltrating CD4+ Tconv cells and CD8+ T cells as well as splenic Treg cells, CD4+ Tconv cells and CD8+ T cells. Overall design: We isolated regulatory and conventional CD4+ T cells as well as CD8+ T cells from B16 melanoma and spleen of mice. Cells were isolated, pooled and target cells were purified by FACS. RNA was extracted and gene expression measured.
Tryptophan metabolism is a key process that shapes the immunosuppressive tumor microenvironment. The two rate-limiting enzymes that mediate tryptophan depletion, indoleamine-2,3-dioxygenase (IDO) and tryptophan-2,3-dioxygenase (TDO), have moved into the focus of research and inhibitors targeting IDO and TDO have entered clinical trials. Local tryptophan depletion is generally viewed as the crucial immunosuppressive mechanism. In T cells, the kinase general control non-derepressible 2 (GCN2) has ...[more]
Project description:miRNA expression profiling in highly purified murine CD4+ Tconv and Treg cells. FoxP3-GFP-hCre1a(high) reporter mice were used to separate both populations based on surface markers and presence or absence of GFP. Two-condition experiment, Tconv vs. Treg. Biological replicates: 1 Tconv, 1 Treg, purified from the same pooled mice. One replicate on 1 array.
Project description:A comparative analysis of gene expression of CD4+ EGFP+ Nrp1+ (tTreg, thymus-derived Treg), CD4+ EGFP+ Nrp1- (pTreg, peripherally-derived Treg) and CD4+ EGFP- (Tconv, conventional T cell) in CD28-/- Foxp3EGFP and Foxp3EGFP mice. Overall design: Nrp1+ Treg (tTreg), Nrp1- Treg (pTreg) and Tconv were sorted from Foxp3EGFP and CD28-/-Foxp3EGFP mice. Total RNAs were extracted from whole samples and analyzed by RNA-seq.
Project description:Distinct populations of progenitor exhausted (Tcf1+Tim-3-) and terminally exhausted (Tcf1-Tim-3+) CD8+ T-cells occur in B16-OVA tumors Overall design: Profiling of CD8+ T-cells from day 10 and day 20 B16-OVA mouse melanoma tumors
Project description:Both Nr4a family nuclear orphan receptors and Foxp3 had been revealed to be crucial transcription factors in Treg cell development. In this study, to reveal their roles in a Treg cell developmental transcriptional programs, we compared transcriptomes among wild-type conventional CD4 T (Tconv) cells, wild-type Treg cells, Nr4a-triple-knockout (Nr4a-TKO) Treg precursor (preTreg) cells, and Foxp3-KO preTreg cells by microarray. Overall design: Total RNA was extracted from wild-type Tconv (CD4-single positive (CD4SP),CD25-low) thymocytes, wild-type thymic Treg (CD4SP, Foxp3+), Nr4a-TKO preTreg (CD4SP, CD25-high, GITR-high, Nr4a3-high), and Foxp3-KO preTreg (CD4SP, CD25high, Foxp3-reporter+) cells. Each cell population was analyzed with two biological replicates.
Project description:We reported transcriptional characterization of Treg and Tconv cells from thymic, splenic, and visceral adipose tissue (VAT) of vTreg53 TCR transgenic mice and control littermates. We examined the effect of Foxp3 on splenic and VAT CD4+ T cell transcriptome. We profiled gene expression in a novel PPARg+ splenic Treg population. We uncovered that the characteristic phenotype of VAT Treg cells was acquired in two stages. Overall design: Gene expression profiles of thymic, splenic, VAT Treg, Tconv, and Foxp3-transduced Tconv cells from vTreg53 TCR transgenic and PPARg reporter mice.
Project description:Microarray used to detail the global gene transcription underlying sorted IFNg+ and IFNg- Tregs (CD4+CD25+CD127lo) and Tconv (CD4+CD25-CD127+) for fresh (unexpanded) and 14 day expanded cells from human blood. Treg and Tconv were FACS isolated from five healthy subjects (median age of 26, range 22-30). Sorted cells were separated into two groups: the first group was stimulated for 4 hours with PMA/ionomycin and labeled with the IFNg cytokine capture kit (Miltenyi Biotech) followed by FACS isolation of IFNg- and IFNg+ populations. The second set was expanded to day 14 prior to reactivation and cytokine cell capture. For each IFNg sorted population, cells were pelleted and flash frozen before RNA isolation and processing.
Project description:FoxP3 is a central regulator of immunological tolerance, controlling the development and function of regulatory T (Treg) cells. To dissect the complex processes orchestrated by FoxP3, we investigated impacts of three autoimmune disease-associated missense FoxP3 mutations (i.e., I363V, A384T, R397W) through knock-in mutagenesis in mice. We investigated the impacts of these mutations on Treg cell transcriptome by microarray analysis. Overall design: CD4(+)hCD2(+) WT or mutant Treg cells were sorted from FoxP3WT or mutant:hCD2/WT:GFP heterozygous mice. CD4(+)GFP(+) WT Treg cells were also sorted from FoxP3WT or A384T:hCD2/WT:GFP mice. CD4(+)hCD2(-)GFP(-) Tconv cells were also obtained from FoxP3WT:hCD2/WT:GFP mice. All cell populations analyzed were generated in duplicates or triplicates.
Project description:This SuperSeries is composed of the following subset Series: GSE11580: Time course RA-treatment of B16 mouse melanoma cells GSE11584: Melan-a mouse melanocytes vs. B16 mouse melanoma cells Keywords: SuperSeries Refer to individual Series
Project description:Co-stimulatory molecules of the CD28 family on T lymphocytes integrate cues from innate immune system sensors, and modulate activation responses in conventional CD4+ T cells (Tconv) and their FoxP3+ regulatory counterparts (Treg). To better understand how costimulatory and co-inhibitory signals might be integrated, we profiled the changes in gene expression elicited in the hours and days after engagement of Treg and Tconv by anti-CD3 and either anti-CD28, -CTLA4, -ICOS, -PD1, -BTLA or -CD80. Total CD4+ T cells were stimulated by anti-CD3 and either anti-CD28, -CTLA4, -ICOS, -PD1, -BTLA or -CD80 antibodies for 1, 4, 20 and 48 hrs and Tconv and Treg were separated by flowcytometry. The 1 and 4 hr lysates were pooled [the 'early' samples] before RNA purification and profiling, as were the 20 and 48 hr samples [the 'late' samples] (note; for Treg cells, only the 20 hr sample was used). RNA was amplified, labeled and hybridized to Mouse Gene 1.0 ST arrays with the data generation and quality control pipeline of 19 the Immunological Genome Project (www.immgen.org), in biological triplicates (duplicates only for ICOS and CD80). Raw data were background-corrected and normalized using the RMA algorithm.
Project description:We used microarray analysis to compare miRNA expression profiles of novel type of Treg cell line, termed HOZOT, with those of activated T cells, so called conventional CD4+ T (Tconv) cells. Overall design: HOZOT-17, a representative HOZOT cell line, was used for profiling. Tconv cells were prepared from the same UCB source as HOZOT-17. CD4+ cells were activated with anti-CD3 antibody plus anti-CD28 antibody (CD3/CD28) and cultured for at least seven days in the presence of IL-2. The chip used for this analysis was a mirVanaTM miRNA Bioarray system.