Project description:During inflammation, neutrophils are rapidly mobilized from the bone marrow storage pool into peripheral blood to subsequently enter lesional sites where most of them rapidly undergo apoptosis. Monocytes constitute a second wave of inflammatory immigrates giving rise to long-lived macrophages and dendritic cell subsets. According to descriptive immunophenotypic and cell culture studies, neutrophils may directly M-bM-^@M-^\transdifferentiateM-bM-^@M-^] into monocytes/macrophages. We here provide mechanistic data in human supporting the existence of this cellular pathway. Global transcriptional profiling of neutrophil-dervived monocytic cells revealed close resemblance of gene expression with monocytes and a clear separation of these cells from neutrophils. G-CSF mobilized neutrophils from peripheral blood were isolated and in vitro stimulated with GM-CSF, TNFa, and Il-1b for 5 days to generate monocytic cells. RNA were isolated from these cells and freshly isolated blood neutrophils and monocytes.
Project description:During inflammation, neutrophils are rapidly mobilized from the bone marrow storage pool into peripheral blood to subsequently enter lesional sites where most of them rapidly undergo apoptosis. Monocytes constitute a second wave of inflammatory immigrates giving rise to long-lived macrophages and dendritic cell subsets. According to descriptive immunophenotypic and cell culture studies, neutrophils may directly “transdifferentiate” into monocytes/macrophages. We here provide mechanistic data in human supporting the existence of this cellular pathway. Global transcriptional profiling of neutrophil-dervived monocytic cells revealed close resemblance of gene expression with monocytes and a clear separation of these cells from neutrophils.
Project description:Neutrophils provide immune protection against pathogens but also may promote tissue injury in inflammatory diseases. Although neutrophils are generally considered as a relatively homogeneous population, evidence for heterogeneity is emerging. Under steady-state conditions, neutrophil heterogeneity may arise from ageing and the replenishment by newly released neutrophils from the bone marrow. We used microarray to globally analyze gene expression in aged neutrophils and characterize the inflammatory programs that are activated during the aging process in the circulation. Control, aged and activated neutrophils were sorted directly from mouse blood for RNA extraction and hybridization on Affymetrix microarrays. We transfused whole blood and harvested donor neutrophils marked by the CD45.1 allele 6h later to derive neutrophils that had truly aged in vivo. Sorted neutrophils were compared to control donor neutrophils that had been transferred for only 10 min. Additionally, we harvested circulating neutrophils from TNF-? treated mice for comparison with neutrophils activated by systemic inflammation.
Project description:The purpose of this study is to learn the dynamic transcriptomic and molecular changes during neutrophil development. We investigated the gene expression profiles of purified morphology-defined neutrophil populations. Promyelocytes (PMs), myelocytes (MCs), metamyelocytes (MMs), band neutrophils and segmented neutrophils (BN/SNs) were isolated by FACS based on differential expression of C-Kit and Ly6g.Their identities were confirmed by morphological examination. Bulk RNA sequencing was then performed to reveal the gene expression profiles and molecular signatures of each cell population. Detailed analysis of gene expression revealed that MBs highly expressed the stem cell marker Cd34 and translation-related genes such as Eef1a1, while the highest expression of primary granule-related genes such as Mpo, Elane, Prtn3, and Cstg was detected in the PM population. MCs and MMs were highly proliferative neutrophils expressing high levels of cell cycle-related genes which were significantly downregulated in mature neutrophils. Finally, this study provides a framework for the detail study of the neutrophil development and for the future application in the modulation of the granulopoiesis under pathological conditions.
Project description:To investigate the role of DREAM (a transcriptional repressor) in regulating gene transcription in neutrophils under inflammatory conditions, we conducted the next generation sequencing using unstimulated and TNF-alpha-stimulated neutrophils isolated from WT and DREAM KO mice.
Project description:Neutrophils provide immune protection against pathogens but also may promote tissue injury in inflammatory diseases. Although neutrophils are generally considered as a relatively homogeneous population, evidence for heterogeneity is emerging. Under steady-state conditions, neutrophil heterogeneity may arise from ageing and the replenishment by newly released neutrophils from the bone marrow. We used microarray to globally analyze gene expression in aged neutrophils and characterize the inflammatory programs that are activated during the aging process in the circulation.
Project description:To explore the mechanism related to the induction of monocytic differentiation in AML combination of ATRA and RAD001, gene expression patterns of THP-1 cells were analyzed using the Agilent Whole Human Genome Expression Array. Gene expression patterns of THP-1 cell cultured under four different conditions were analyzed. The conditions were as follows; 1, untreated. 2, treated with ATRA (1mM) and RAD001 (2.5nM). Total RNA was extracted at 5 days after starting treatment.