Project description:These data are part of a body of work exploring the effect of IgG from patients with ANCA vasculitis on human monocytes in vitro. Please see the relevant publication for a full description.
Project description:To screen the genes and pathways regulated by CREB1 in the THP1 monocytes induced by LPS, THP1 monocytes were transfected by CREB1 shRNA and then treated by LPS for 3 hours.
Project description:The etiopathogenesis underlying myeloperoxidase anti-neutrophil cytoplasmic antibody associated glomerulonephritis (MPO-AAGN) remains incompletely understood. Furthermore, there are only limited treatment options and treatment resistance of MPO-AAGN is still a common problem. To identify new targeted treatment options, intrarenal single-cell RNA sequencing (scRNA-seq) was applied to kidney biopsies from MPO-AAGN patients and control health kidney tissues to define the transcriptomic landscape at single-cell resolution. Intrarenal scRNAseq was also applied to a pre-clinical mouse model of MPO-AAGN to show that this model of disease can be used to trial new targeted treatments. NF-κB pathway activation was confirmed in a variety of kidney cells in MPO-AAGN patients. Kidney infiltrating immune cells of MPO-AAGN patients were mainly enriched in inflammatory pathways including TNF signaling, IL-17 signaling and NOD-like receptor signaling. These findings were similar in our pre-clinical mouse model of MPO-AAGN. Furthermore, there was an overexpression of inflammasome related genes (AIM2, IFI16) in MPO-AAGN patients. A dynamic gene expression in glomerular resident cells was observed in MPO-AAGN, including increased expression of several genes, including CD9 and SPARC, which were closely related to parietal epithelial hyperplasia and crescent formation and lesion progression. Importantly, overexpression of HSP90AA1 in non-focal mesangial cells and endothelial cells was found and the expression of several chemokines (CCL20, CXCL3, CXCL8, CXCL1, CCL2) were upregulated in non-focal proximal tubule cells. Moreover, MPO-AAGN patients with treatment resistance had higher proportions of kidney infiltrating classical monocytes and CD8+ T cells. Elevated expression of SPARC and LAMA4 in mesangial cells, IL33 in endothelial cells, and CFL1 in several cell clusters (proximal tubule cells, loop of Helen, macrophages) were observed in MPO-AAGN patients with treatment resistance when compared with patients who achieved remission after induction therapy. These results offer new insight into the pathogenesis of the progression and treatment resistance MPO-AAGN. We have identified new therapeutic targets for MPO-AAGN that can be tested in a pre-clinical model of disease.
Project description:To screen the genes regulated by BRD3 in the THP1 monocytes induced by LPS, THP1 monocytes were treated by LPS for 1 hour in the presence or absence of OTX015
Project description:CD14+ Monocytes from healthy volunteers were purified by MACS (negative selection) and FACSorting and either left untreated or stimulated for 24h and 48h with LPS. THP-1 cells were stimulated for 4h, 24h and 48h with LPS. Glycoproteins were captured with hydrazide chemistry and tryptic and PNGase F-released peptide fractions analyzed by MS/MS. Quantitative assessment revealed differential glycoprotein expression in activated/LPS-tolerized monocytes and naïve monocytes and THP-1 cells.
Project description:In vitro priming of human peripheral monocytes with the filarial extract from Brugia malayi and subsequent LPS stimulation to explore the immunmodulatory capacity of filarial extract to subsequent inflammatory LPS responses and thus to identify new candidates to modify TLR4-associated responses/diseases and filarial pathology.
Project description:To investigate the possible therapeutic effects of ibuprofen and Pep19-2.5 alone or in combination on the LPS-response of human monocytes. Human monocytes were challenged with 0.1ng/ml LPS and/or 10ng/ml Pep19-2.5 LPS. Medium controls served as unstimulated samples. LPS and Pep19-2.5 was used either alone or together to LPS-challenged samples. RNA was extracted after 4 hrs of stimulation. Three biological replicates were conducted
Project description:The heterogeneous characteristics of activated monocytes are much unknown. We performed the single cell transcriptomic analysis of LPS-treated human monocytes from healthy blood donor, and CD127 expression associated expression pattern of inflammatory genes was identified in LPS-treated monocytes.
Project description:To investigate the possible therapeutic effects of ibuprofen and Pep19-2.5 alone or in combination on the LPS-response of human monocytes.