Project description:We used microarrays to detail the global gene expression changes elicited by stimulation of human peripheral blood monocytes from buffy coats with 50µM Annexin A1 peptide Ac1-25 for 2h versus unstimulated control monocytes. Keywords: comparative gene expression study
Project description:We used microarrays to detail the global gene expression changes elicited by stimulation of human peripheral blood monocytes from buffy coats with 50µM Annexin A1 peptide Ac1-25 for 2h versus unstimulated control monocytes. Experiment Overall Design: three independent monocyte isolations each were used for stimulated and control cell RNA preparations and subsequent hybridization to arrays according to manufacturers protocols
Project description:CD14+ monocytes were separated from human peripheral blood and exposed to IL-4 for 12 or 72 hours then subjected to microarray analysis We used Affymetrix miRNA1.0 microarray to obtain global miRNA expression data of human monocytes, unstimulated and IL-4-stimulated differentiating macrophages.
Project description:The glucocorticoid-inducible protein annexin A1 has been shown to function as key-regulator of the resolution phase of inflammation but its role in immune-mediated crescentic glomerulonephritis has not been studied so far. Acute crescentic glomerulonephritis was induced in annexin A1 deficient and wildtype mice using a sheep serum against rat glomerular basement membrane constituents. Alterations in gene expression were determined by RNA-Seq and gene ontology analysis. Intrinsic annexin A1 has a protective effect in reducing pro-inflammatory signals and infiltration of neutrophil granulocytes during crescentic GN. The annexin A1 signaling cascade may therefore provide novel targets for the treatment of inflammatory kidney disease.
Project description:Rationale: Changes in peripheral blood cell populations have been observed but not detailed at single-cell resolution in idiopathic pulmonary fibrosis (IPF). Objectives: To provide an atlas of the changes in the peripheral immune system in stable and progressive IPF. Methods: Peripheral blood mononuclear cells (PBMCs) from IPF patients and controls were profiled using 10x Chromium 5’ single-cell RNA sequencing (scRNA-seq). Flow cytometry was used for validation. Protein concentrations of Regulatory T-cells (Tregs) and Monocytes chemoattractants were measured in plasma and lung homogenates from patients and controls. Measurements and Main Results: Thirty-eight PBMC samples from 25 patients with IPF and 13 matched controls yielded 149,564 cells that segregated into 23 subpopulations, corresponding to all expected peripheral blood cell populations. Classical monocytes were increased in progressive and stable IPF compared to controls (32.1%, 25.2%, 17.9%, respectively, p<0.05). Total lymphocytes were decreased in IPF vs controls, and in progressive vs stable IPF (52.6% vs 62.6%, p=0.035). Tregs were increased in progressive IPF (1.8% vs 1.1%, p=0.007), and were associated with decreased survival (P=0.009 in Kaplan-Meier analysis). Flow cytometry analysis confirmed this finding in an independent cohort of IPF patients. Tregs were also increased in two cohorts of lung scRNA-seq. CCL22 and CCL18, ligands for CCR4 and CCR8 Treg chemotaxis receptors, were increased in IPF. Conclusions: The single-cell atlas of the peripheral immune system in IPF, reveals an outcome-predictive increase in classical monocytes and Tregs, as well as evidence for a lung-blood immune recruitment axis involving CCL7 (for classical monocytes) and CCL18/CCL22 (for Tregs).
Project description:Human peripheral blood monocytes were treated with control or with 25 ng/ml IFNg for 24 hours. RNA was collected, processed and hybridized to Affymetrix HGU133Plus2 chips.
Project description:Monocytes were isolated from blood of Wildtype, Mir150 and TET3 knockout mice, sorted for classical and non-classical monocyte subsets. RNA seq was performed on unstimulated monocytes.
Project description:To investigate the effect of ANXA1, encoding annexin a1, on group 2 innate lymphoid cells (ILC2s), we downregulated the expression using lentiviral vector-based shRNA transfer and performed RNA-seq analysis.
