Project description:Until today, natural Echinacea purpurea extracts are widely used to prevent and treat respiratory trac infections. Although its immunomodulatory effects have already been demonstrated by several studies, the molecular mechanisms involved are not yet completely understood. In this study, we applied a systems biology approach by integrating transcriptome, kinome and methylome profiles of THP-1 monocyte cells treated with Echinaforce®, a commercial standardized ethanolic tincture of Echinacea purpurea. The latter induced the expression of various genes linked to the anti-viral interferon pathway, NF-κB signaling and chemotaxis. In addition, kinase activity profiling showed that Echinaforce® increased JAK1 kinase activity, which is crucial in the activation of the JAK-STAT pathway to induce interferon-stimulated genes and MAPK kinase activity. Furthermore, gene expression changes following Echinaforce® treatment occurred largely independent from DNA methylation changes. Of special note, Echinaforce® induced global DNA hypermethylation in gene bodies, intergenic, and CpG-poor repeat regions, which could be part of an evolutionary conserved anti-viral response to suppress viral integration and/or replication. Altogether, we demonstrate that Echinaforce® treatment promotes an anti-viral response in THP-1 cells through activation of interferon, MAPK and NF-κB signaling pathways and DNA hypermethylation of repeat elements.
Project description:Until today, natural Echinacea purpurea extracts are widely used to prevent and treat respiratory trac infections. Although its immunomodulatory effects have already been demonstrated by several studies, the molecular mechanisms involved are not yet completely understood. In this study, we applied a systems biology approach by integrating transcriptome, kinome and methylome profiles of THP-1 monocyte cells treated with Echinaforce®, a commercial standardized ethanolic tincture of Echinacea purpurea. The latter induced the expression of various genes linked to the anti-viral interferon pathway, NF-κB signaling and chemotaxis. In addition, kinase activity profiling showed that Echinaforce® increased JAK1 kinase activity, which is crucial in the activation of the JAK-STAT pathway to induce interferon-stimulated genes and MAPK kinase activity. Furthermore, gene expression changes following Echinaforce® treatment occurred largely independent from DNA methylation changes. Of special note, Echinaforce® induced global DNA hypermethylation in gene bodies, intergenic, and CpG-poor repeat regions, which could be part of an evolutionary conserved anti-viral response to suppress viral integration and/or replication. Altogether, we demonstrate that Echinaforce® treatment promotes an anti-viral response in THP-1 cells through activation of interferon, MAPK and NF-κB signaling pathways and DNA hypermethylation of repeat elements.
Project description:SP110b is an interferon (IFN)-induced nuclear protein and may function as a transcriptional co-activator/repressor. IFNγ activates monocytes/macrophages thereby mediating inflammation. However, uncontrolled activation induces monocyte/macrophage cell death, which may cause immunopathology. We have demonstrated that SP110b expression prevented IFNγ-mediated monocyte/macrophage cell death. To explore the molecular mechanisms by which SP110b suppresses IFNγ-induced cell death, we performed a genome-wide microarray analysis to identify genetic determinants associated with IFNγ-induced cell death and regulated by SP110b. We sought to identify genetic determinants associated with IFNγ-induced cell death and regulated by SP110b. To that end, THP1 human monocyte-like cells that could be induced by doxycycline (Dox) to over-express SP110b (THP1-SP110b) were generated and 5 experimental groups of THP1-SP110b cells were harvested for RNA extraction and hybridization on Affymetrix microarrays. The 5 groups are as follows: untreated THP1-SP110b cells as control (CON), cells treated with IFNγ for 2 days (IFN_2D), cells treated with Dox plus IFNγ for 2 days (DoxIFN_2D), cells treated with IFNγ for 4 days (IFN_4D), and cells treated with Dox plus IFNγ for 4 days (DoxIFN_4D).
Project description:Macrophages are sentinels of the immune system and THP1 monocytic cells are one of the widely used models to study immune responses in macrophages. Several monocyte-to-macrophage differentiation protocols exist with phorbol 12-myristate-13-acetate (PMA) being widely used and accepted. However, the concentrations and durations of PMA treatment to induce differentiation varies widely in published literature. In this study, we determined the proteome expression dynamics of THP1 macrophages differentiated from monocytes by three commonly used PMA-based differentiation protocols using dimethyl labeling-based quantitative proteomics analysis. Our analysis shows that variations in PMA concentration and varying periods of rest post-stimulation result in downstream differences in proteome expression and cellular processes. We demonstrate that these differences result in altered expression of cytokines upon stimulation with different TLR ligands. Together, these findings provide a valuable resource that significantly expands the knowledge of protein expression dynamics in in vitro models of macrophages which in turn has profound impact on the immune responses being studied.
Project description:We used an inducible shRNA system and RNA-Seq to examine gene expression changes in acute myeloid leukemia THP1 cells following silencing of RUVBL2. RUVBL2 is a AAA+ ATPase that functions in a number of cellular processes, including chromatin remodeling and trnascriptional control, and is critical for survival of acute myeloid leukemia cells and in vivo disease progression.
Project description:In order to further explore the expression of lncrna in asthma, we used lipopolysaccharide (LPS) to activate human monocyte macrophage THP-1, and then used deep sequencing method to detect the expression of lncrna in activated and inactive THP1 cells. The results showed that there were different expression of lncrna in LPS activated and inactive THP1 cells