Project description:The goal of this study was to compare gene expression in mouse bone marrow-derived macrophages stimulated with high or low doses of the TLR4 ligand Kdo2-Lipid A.
Project description:Gene expression for mouse C57BL/6 bone marrow derived macrophages stimulated with a titration matrix of TLR4 and TLR2 ligand, KDO2-Lipid A and Pam3CSK4, respectively
Project description:The model was constructed to describe TLR4 induced NF-κB activation in native bone marrow-derived macrophages. It included processes of ligand (lipopolysaccharide) recognition, formation of dimer receptor complex and further signal transduction through TRAF6/TAK1 complex that leads to the activation of IKKα/β kinase, which in turn enables the NF-κB transcription factor phosphorylation and translocation in the cell nucleus, and induction of IkB and WIP1 (as an example of induced protein that promotes NF-κB dephosphorylation 2) gene transcription. Models were based on the current knowledge of TLR signaling framework, protein interactions within the TLR4 pathway, and up-to-date mathematical models describing Toll receptor activation.
The major important additions were made to TLR4 signaling description:
1) Receptor dimerization process
2) The existence of a basal nuclear NF-κB level (translocation)
3) NF-κB phosphorylation by IKK complex
Project description:WT control or IFNAR deficient bone marrow derived macrophages were stimulated with IFNb, TLR3, IFNg, TLR4, and TLR4+IFNg ligands for 48 h.
Project description:Transcriptomic analysis of the temporal changes induced in mouse bone marrow derived macrophages (BMDMs) by the cytokine Interferon-beta over a timecourse of 0 to 24 hours of treatment. We set out to study the transcriptional events in mouse macrophages over time following stimulation with Interferon-beta. Mouse bone marrow derived macrophages were stimulated for 1, 2, 4, 8 and 24 hours with 10U/mL mouse interferon-beta or left untreated.
Project description:Chronic, low-grade inflammation has a widespread and significant impact on health, especially in Western-society. While inflammation is beneficial for the removal of microbes, low-grade inflammation never resolves and can cause or worsen other diseases. However, the process by which low-grade inflammation occurs is poorly understood. As lipopolysaccharide (LPS) is associated with chronic inflammatory diseases, we exposed murine bone-marrow derived monocytes to chronic LPS-stimulation at Low-dose (100pg/mL) or High-dose (1 µg/mL), as well as a PBS control. The cells were profiled for H3K27ac expression and gene expression. The gene expression of TRAM-deficient and IRAK-M-deficient mice with LPS exposure was also analyzed for mechanistic insight. We determined that many of the differences between the Low-dose and High-dose conditions are related to the TRIF-dependent pathway of TLR4 signaling. Furthermore, these changes are also seen in the epigenome, suggesting the epigenome may be what leads to the differential response. These findings further characterize the different means utilized in low-grade conditions, and how it might lead to a damaging, non-resolving state. Moreover, our data provide potential targets for future mechanistic or therapeutic studies.
Project description:WT control or MyD88 deficient bone marrow derived macrophages were stimulated with TLR2, TLR3, TLR4, TLR7, and TLR9 ligands for 48 h.
