Project description:Expression profiling of HCT116 colon cancer cells with different treatments. Illumina HumanHT-12 V4.0 expression beadchip was used to obtain expression profiles across more than 31,000 annotated genes. Total RNA obtained from HCT116 cells with different treatments, namely, cells with TNF-α (50ng/ml)/IFN-ɣ (50U/ml) treatment, cells with TNF-α (50 ng/ml)/IFN-ɣ (50 U/ml) and silibinin (100 uM) treatment and cells without any treatment. Each sample has 2 biological replicates.
Project description:G-protein coupled receptors (GPCRs) have diverse roles in physiological processes, including immunity. Gs-coupled GPCRs increase while Gi-coupled ones decrease intracellular cAMP. Previous studies suggest that, in epithelial cells, Gs-coupled GPCRs enhance whereas Gi-coupled GPCRs suppress pro-inflammatory immune responses. In order to examine the issue, we chose beta2 adrenergic receptor and GPR40 as representatives of Gs- and Gi- coupled GPCRs, respectively, and examined their effects on TNF-alpha and IFN-gamma-(TNF-alpha + IFN-gamma) induced gene expression by HaCaT. We used microarrays to detail the global changes of gene expression induced by a beta2 adrenergic receptor agonist terbutaline or GPR40 agonist GW9508 pre-treatment in TNF-alpha + IFN-gamma - stimulated HaCaT cells. HaCaT cells were pre-treated with terbutaline or GW9508, TNF-alpha + IFN-gamma were then added, and cultured for another 24 h. Cells were then used for RNA extraction and hybridization on Affymetrix microarrays. We sought to clarify changes in gene expression after 1) TNF-alpha + IFN-gamma, 2) TNF-alpha + IFN-gamma + terbutaline, and 3) TNF-alpha + IFN-gamma + GW9508 treatment. To this end, we set 4 groups of samples; 1) unstimulated group, 2) TNF-alpha + IFN-gamma-stimulated group, 3) TNF-alpha + IFN-gamma + terbutaline-stimulated group, and 4) TNF-alpha + IFN-gamma + GW9508-stimulated group. In each group, HaCaT cells were stimulated in triplicate wells (n=3).
Project description:We have previously reported that the dengue virus (DENV) type 3 P12/08 strain caused a lethal systemic infection, severe vascular leakage at terminal stage in IFN-α/β and γ receptors knockout mice (IFN-α/β/γRKO mice), and blockade of TNF-α signaling drastically protected mice. However, the detailed pathological mechanism remains unknown. Therefore, we performed transcriptome analysis of liver and intestinal specimens, which showed most clearly exhibited vascular leakage, chronologically collected from infected- IFN-α/β/γRKO mice with/without anti-TNF-α Ab treatment.
Project description:Genome wide DNA methylation profiling of CD11b+CD33+ cells after treatment with different cytokines. The Illumina Infinium MethylationEPIC Beadchip was used to obtain DNA methylation profiles across approximately 850,000 CpGs in CD11b+CD33+ cells isolated from PBMCs treated in vitro for 4 days with TNF-alpha, IFN-alpha, IFN-gamma or control.
Project description:Analysis of MIN6 murine beta cell line transfected with ARH3 RNAi and treated with pro-inflammatory cytokines TNF-alpha, IL-1beta and IFN-gamma.
Project description:Analysis of MIN6 murine beta cell line transfected with Pla2g6 RNAi and treated with pro-inflammatory cytokines TNF-alpha, IL-1beta and IFN-gamma.
Project description:Exosomes are extracellular vesicles that function in intercellular communication. We have previously reported that exosomes play a role in the transmission of antiviral molecules during interferon-α (IFN-α)-mediated immune responses. In this study, the protein contents of THP-1-derived macrophages with or without interferon-α treatment and of the exosomes secreted from these cells were analyzed by the label-free LC-MS/MS quantitation technologies. A total number of 1845 and 1550 protein groups were identified in the THP-1 macrophages and the corresponding exosomes, respectively. Treating the cells with IFN-α resulted in the differential abundance of 110 proteins in cells and 260 proteins in exosomes (greater than 2.0-fold), among which 35 proteins were both up-regulated in the IFN-α treated cells and corresponding exosomes while 139 proteins were specifically up-regulated in exosomes but not in the donor cells. GO and KEGG analysis of the protein function categories suggested that IFN-α promoted the abundance of proteins involved in “defense response to virus” in both exosomes and cells, and proteins related to “RNA processing” only in exosomes. Functional analysis further indicated that exosomes from IFN-α-treated cells exhibited potent antiviral activity that restored the impaired antiviral response of IFN-α in hepatitis B virus-replicating hepatocytes. These results have deepened the understanding of the exosome-mediated transfer of IFN-α-induced antiviral molecules and may provide new basis for therapeutic strategies to control viral infection.
Project description:To investigate effects of 4 mixture of cytokines including TNF-alpha, IL-1beta IFN-gamma, and TGF-bata1 on rhematoid arthritis synovial fibroblasts (RASFs), we treated RASFs (n = 3) with TNF-alpha (1 ng/mL), IL-1beta (0.1 ng/mL), IFN-gamma (10 ng /mL), TGF-beta1 (1 ng/mL) (4mix) or vehicle control (Veh) for 24 hours and conducted RNA-seq analysis.
Project description:To investigate effects of TEAD4 knockdown on rhematoid arthritis synovial fibroblasts (RASFs) stimulated with 4 mixture of cytokines including TNF-alpha, IL-1beta IFN-gamma, and TGF-bata1 , we pre-treated RASFs (n = 3) with siControl (siCtrl) or siTEAD4 for 72 hours, and then treated them with TNF-alpha (1 ng/mL), IL-1beta (0.1 ng/mL), IFN-gamma (10 ng /mL), TGF-beta1 (1 ng/mL) (4mix) for 24 hours and conducted RNA-seq analysis.
