Project description:We report the use of bulk RNAseq of AML and endothelial cell lines in four conditions (1. Untreated; 2. IFNg - 10 ng/ml; 3. TNFa - 10 ng/ml; 4. IFNg and TNFa - 10 ng/ml) in order to identify genes that are differentially expressed between the two cell types, both at baseline and in an inflammatory microenvironment.
Project description:The psoKC (psoriatic keratinocyte) model is represnting the behavour of keratinocytes in the later or chronic stage of psoriasis in response to the main cytokines that constitute the characteristic cytokine milieu, namely IFNg and TNFa (mainly derived by Th1 cells), and IL-17 and IL-22 (mainly derived by Th17 cells).
Additionally, the model explores the role of exogenous PGE2 through the activation of EP4 receptor signaling. The response to the aforementioned stimuli was not only limited to the cell fate decisions of keratinocytes (proliferation, apoptosis or differentiation) but also include their effect on the psoriatic environment with respect to the secretion of ligands and intercellular-acting stimuli.
Project description:The study sought to determine the global miRNA profiles of mouse pancreatic cell lines aTC1-6 and bTC1 at steady state and after treatment with a cocktail of pro-inflammatory cytokines (IL1b; IFNg and TNFa) for a time course of 24 and 48 hours. Inflammatory cocktail contains IL1b 50 IU/ml; IFNg 1000 IU/ml; TNFa 1000 IU/ml. MicroRNA expression profiles in both cell lines during the different experimental conditions were determined using TaqMan® Rodent miRNA A+B Cards Set v2.0 (TLDA, Life Technologies). MicroRNA profiles were measured in 3 independent biological replicates.
Project description:Beta cells intrinsically contribute to the pathogenesis of type 1 diabetes (T1D), but the genes and molecular processes that mediate beta cell survival in T1D remain largely unknown. We combined high throughput functional genomics and human genetics to identify T1D risk loci regulating genes affecting beta cell survival in response to the proinflammatory cytokines IL-1b, IFNg, and TNFa. We mapped cytokine-responsive candidate cis-regulatory elements (cCREs) active in beta cells using ATAC-seq and single nuclear ATAC-seq (snATAC-seq), and linked cytokine-responsive beta cell cCREs to putative target genes using single cell co-accessibility and HiChIP. We performed a genome-wide pooled CRISPR loss-of-function screen in EndoC-βH1 cells, which identified hundreds of genes affecting cytokine-induced beta cell loss. We identified thousands of variants in cytokine-responsive beta cell cCREs altering transcription factor (TF) binding using high-throughput SNP-SELEX. Together our findings reveal processes and genes acting in beta cells during cytokine exposure that intrinsically modulate risk of T1D.
Project description:To investigate Tr1 cells and TNFa single expressing cells we stimulated PBMCs from primigravid women with VAR2CSA expressing P. falciparum and sorted CD4+ T cells expressing both IL10 and IFNg or only TNF
Project description:Combinatorial cytokine signaling is implicated in the pathogenesis of diseases of the cardiovascular system ranging from acute combinatorial cytokine signaling observed in complications from SARS-COVID19 infection to chronic combinatorial cytokine signaling observed in atherosclerosis. There remains a large gap of our understanding how combinatorial cytokine signaling interface to drive proinflammatory programs. We hypothesize that stimulation of HAECs with IFNg and TNFa promotes synergistic gene transcription through the coordinated interplay between p65, STAT1, p300/CBP, and BRD4. Our work demonstrates through the generation of novel statistical framework that IFNg and TNFa dual stimulation for one hour results in the synergistic induction of a small subset of proinflammatory genes. Prolonged dual cytokine stimulation results in synergistic levels of protein expression and cell death. Treatment with A-485 and/or JQ1 reduces levels of synergistic gene and protein expression along with improvements in cell viability. Defining synergistic responses and identifying their unique transcriptional dependencies opens the door to understanding how disparate cytokine signaling drives proinflammatory responses and how these responses can be targeted.
Project description:Stromal fibroblasts reside in inflammatory tissues that are characterized by either immune suppression or activation. Whether and how fibroblasts adapt to these contrasting microenvironments remains unknown. Cancer-associated fibroblasts (CAFs) mediate immune quiescence by producing the chemokine CXCL12, which coats cancer cells to suppress T cell infiltration. We examined whether CAFs can also adopt an immune-promoting chemokine profile. Single-cell RNA-sequencing of CAFs from mouse pancreatic adenocarcinomas identified a sub-population of CAFs with decreased expression of CXCL12 and increased expression of the T cell-attracting chemokine, CXCL9 in association with T cell infiltration. TNFa and IFNg containing conditioned media from activated CD8+ T cells converted stromal fibroblasts from a CXCL12+/CXCL9- immune suppressive phenotype into a CXCL12-/CXCL9+ immune-activating phenotype. Recombinant IFNg and TNFa acted synergistically to induce CXCL9 expression, whereas TNFa alone suppressed CXCL12 expression. This coordinated chemokine switch leads to increased T cell infiltration in an in vitro chemotaxis assay. Our study demonstrates that CAFs have a phenotypic plasticity that allows their adaptation to contrasting immune tissue microenvironments.
Project description:Stromal fibroblasts reside in inflammatory tissues that are characterized by either immune suppression or activation. Whether and how fibroblasts adapt to these contrasting microenvironments remains unknown. Cancer-associated fibroblasts (CAFs) mediate immune quiescence by producing the chemokine CXCL12, which coats cancer cells to suppress T cell infiltration. We examined whether CAFs can also adopt an immune-promoting chemokine profile. Single-cell RNA-sequencing of CAFs from mouse pancreatic adenocarcinomas identified a sub-population of CAFs with decreased expression of CXCL12 and increased expression of the T cell-attracting chemokine, CXCL9 in association with T cell infiltration. TNFa and IFNg containing conditioned media from activated CD8+ T cells converted stromal fibroblasts from a CXCL12+/CXCL9- immune suppressive phenotype into a CXCL12-/CXCL9+ immune-activating phenotype. Recombinant IFNg and TNFa acted synergistically to induce CXCL9 expression, whereas TNFa alone suppressed CXCL12 expression. This coordinated chemokine switch leads to increased T cell infiltration in an in vitro chemotaxis assay. Our study demonstrates that CAFs have a phenotypic plasticity that allows their adaptation to contrasting immune tissue microenvironments.