Project description:Gene expression studies comparing IFNg+ Tregs versus IFNg- Tregs from human peripheral blood Ex vivo sorted Tregs (CD25highCD127neg) were stimulated for 4 hours and IFNg-secreting cells were detected by a IFNg-capture kit. The samples were resorted based on IFNg expression.
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.
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:Human Tregs isolated from PBMCs were sorted into 4 different subpopulations based on IFNg and IL-10 production and were performed mRNA-seq.
Project description:Studying the effect of Th1 cytokine IFNg on gene expression of human neutrophils by comparing gene expression of neutrophiles isolated from the blood of 3 healthy donors and stimulated with IFNg (10ng/ml) or left unstimulated.
Project description:Human peripheral blood monocytes were treated with control or with 25 ng/ml IFNg for 24 hours. RNA was collected, processed and hybridized to Affymetrix HGU133Plus2 chips.
Project description:Tregs in tumors are functionally heterogeneous, and Th1-Tregs represent a highly suppressive subset. This study identifies Tregs themselves as a source of IFN-γ, which promotes Th1-Treg differentiation. Treg-derived IFN-γ acts in an autocrine manner to sustain T-bet expression and the Th1-Treg phenotype, while PF4 from Arg1(+) TAMs further amplifies this pathway by inducing Ifng expression in Tregs. Conditional deletion of Ifng in Foxp3(+) cells impaired Th1-Treg differentiation in both tumors and spleen, and a similar mechanism was observed in EAE. Overall, Treg-derived IFN-γ forms a positive feedback loop with other IFN-γ sources and TAM-derived PF4, driving the maintenance and accumulation of Th1-Tregs and reinforcing immunosuppression.
Project description:Sarcoidosis is characterized by exaggerated immune response to unknown agent and can affect different organs. One of the main players in the pathology of the disease are regulatory T cells (Tregs), however up to date there are no insides on the possible molecular alterations of this particular cell subset. Therefore, in the current study we looked for the global changes on miRNA and mRNA level of Tregs of patients with the most predominant form of the disease - pulmonary sarcoidosis (PS). For this purpose we sorted Treg cells from peripheral blood (PB) and bronchoalveolar lavage (BAL) and analyzed the global molecular changes. MiRNA analysis showed major changes in Tregs isolated from PB to the ones from BAL, pointing at the stronger impact of the microenvironment than the disease state. Additionally, several disease-related miRNAs of PB Tregs were identified like miR-155 and miR-223.
Project description:biomarker discovery pipeline for active and subclinical JIA through 48 gene Treg signature+ nanoString normalised counts from peripheral blood and synovial fluid mononuclear cells and Tregs