Project description:To evaluate gene expression in human peripheral blood derived monocytes over the course of an LPS stimulation time-series. Keywords: time course
Project description:To evaluate gene expression in human peripheral blood derived monocytes over the course of an LPS stimulation time-series. Experiment Overall Design: Blood samples from three individuals, each sample split into separate culture for each timepoint (0, 2, 4, 8, and 24 hours). RNA prepared from all, and analyzed with Bioanalyzer. Two best RNA samples from each timepoint analyzed by hybridization to a chip.
Project description:Inflammatory stimulation with lipopolysachharide (LPS) iduces a strong re-organization in the transcriptional program of human monocytes. We isolated peripheral blood monocytes from neurologically unaffected controls (n=8) and stimulated the monocytes in culture with LPS (1 ng/mL for 4 h) to investigate changes in gene expression upon LPS stimulation.
Project description:This series compares gene expression in chicken peripheral blood lymphocyte (PBL) derived macrophages following stimulation with either XL10 E. coli or S. typh LPS over an 8 hour time course (0,1,2,4,and 8 hours). For the late time points, the stimuli were removed after 2 hours and fresh media was added for the remaining time. Keywords: time-course
Project description:Exposure of human monocytes to lipopolysaccharide (LPS) or other pathogen-associated molecular pattern (PAMPs) induces a temporary insensitivity to subsequent LPS challenges, a cellular state called endotoxin tolerance (ET). The tolerant state of monocytes is accompanied by cell surface and other glycoprotein expression changes induced by the activation of Toll-like receptors (TLRs). In this study, we aimed to characterize the cellular state of human monocytes stimulated with Gram-positive Staphylococcus aureus and TLR2 ligands. We analyzed gene expression changes induced by S. aureus after 2 and 24 hours by amplicon sequencing (RNA-AmpliSeq) and compared the pro-inflammatory response after 2 hours of stimulation to the to the response in re-stimulation experiments after the first stimulus. In parallel, glycoprotein expression changes in human monocytes after 24 hours of S. aureus stimulation were analyzed by proteomics and compared to stimulation experiments with TLR2 ligands Malp-2 and Pam3Cys and TLR4 ligand LPS. The results demonstrate that monocytes stimulated with S. aureus and TLR ligands entered the tolerant cell state after activation. Compared to TLR agonist mediated activation and tolerization of monocytes, glycoprotein expression changes induced by S. aureus stimulation revealed significant differences in receptor expression profiles. We report a glycoprotein expression profile characteristic for PAMP and S. aureus tolerized human monocytes. Finally, we analyzed peripheral blood monocytes of patients with S. aureus bloodstream infection for inflammatory responses in vitro and for their glycoprotein expression profiles. RNA-AmpliSeq data from patient-derived monocytes demonstrated that the cells were pro-inflammatory responsive to S. aureus stimulation and expressed higher level of CD44 mRNA, while other markers of the tolerant cell state were not detected.
Project description:This series compares gene expression in chicken peripheral blood lymphocyte (PBL) derived macrophages following stimulation with either XL10 E. coli or S. typh LPS over an 8 hour time course (0,1,2,4,and 8 hours). For the late time points, the stimuli were removed after 2 hours and fresh media was added for the remaining time.
Project description:HLA-B27-associated inflammatory diseases remains one of the strongest HLA-disease known to date. HLA-B27-associated acute anterior uveitis has wide-ranging medical significance due to its ocular, systemic, immunologic, and genetic features. To investigate the genes and signalling pathways located upstream of the inflammatory processes in HLA-B27-associated acute anterior uveitis will help to know the mechanism of this disease. HLA-B27-positive and -negative monocytes isolated from human peripheral blood were stimulated with Vibrio cholera lipopolysaccharide (LPS). Gene expression microarrays were used to identify the differentially expressed genes, and they were analysed by a series of bioinformatics-based techniques. Gene expression microarray analysis revealed marked differences between B27-positive monocytes in the genes that are upregulated in response to LPS stimulation. Gene Ontology enrichment (GO) and pathway analysis indicated that genes participating in protein transport and folding were essential to the inflammatory process. The LPS receptor, TLR4, induced the Toll-like receptor signalling pathway and pathways related to Vibrio cholerae infection, which are located upstream of the network and contribute to the overall response. Among the DE genes, PIK3CA, PIK3CB, AKT3, and MAPK1 may play critical roles in inflammation.Equivalent LPS stimulation induces a different response in HLA-B27-positive monocytes compared to monocytes lacking this HLA protein, suggesting that the TLR pathway is involved in the pathogenesis of HLA-B27-associated AAU. Blocking this pathway and other pathways by siRNA interference of candidate genes may contribute to the development of a treatment for this type of AAU. HLA-B27-positive monocytes isolated from human peripheral blood were stimulated with Vibrio cholera lipopolysaccharide (LPS) for 12 hours.
Project description:To identify genes in human peripheral blood B cells whose expression changes with acute stimulation through the antigen receptor using anti_IgM. Keywords: time series design
Project description:We performed a transcriptome analysis of M2 macrophages generated in vitro from monocytes isolated from peripheral blood of healthy donors, and studied the effect of miR-221-3p mimic transfection and stimulation with LPS.
