Single-cell RNA sequencing of mouse peripheral blood neutrophils before and one day after corneal injury
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE338556 | GEO | 2026/08/30
REPOSITORIES: GEO
Similar Datasets
Project description:During systemic inflammation, different neutrophil subsets are mobilized to the blood circulation. These neutrophil subsets can be distinguished from normal circulating neutrophils (CD16bright/CD62Lbright) based on either an immature CD16dim/CD62Lbright or a CD16bright/CD62Ldim phenotype. Interestingly, the latter neutrophil subset is known to suppress lymphocyte proliferation ex vivo, but the underlying mechanism is largely unknown. We performed transcriptome analysis on the different neutrophil subsets to identify changes that are relevant for their functions. Neutrophil subsets were isolated by FACS sorting from the blood of healthy volunteers who were administered a single dose of lipopolysaccharide (LPS). The transcriptome was determined by microarray. The mobilized neutrophil subsets were characterized by specific transcriptome profiles reflecting their phase in neutrophil lifespan. Interestingly, the CD16bright/CD62Ldim suppressive neutrophils showed an interferon-induced transcriptome profile. This was confirmed by stimulation of peripheral neutrophils with IFNgamma. These cells acquired the capacity to suppress lymphocyte proliferation through the expression of programmed death ligand 1 (PD-L1). These data demonstrate that the suppressive phenotype of the neutrophil subset is induced by IFNgamma. Specific stimulation of neutrophils might have a pivotal role in regulating lymphocyte-mediated inflammation and autoimmune disease. After LPS infusion, blood was taken at t=0 and t=4 hours. Neutrophils were FACS sorted based on CD16 and CD62L expression. Gene expression of neutrophil subsets was assessed relative to t=0 as control.
2013-05-01 | E-GEOD-42358 | biostudies-arrayexpress
Project description:During systemic inflammation, different neutrophil subsets are mobilized to the blood circulation. These neutrophil subsets can be distinguished from normal circulating neutrophils (CD16bright/CD62Lbright) based on either an immature CD16dim/CD62Lbright or a CD16bright/CD62Ldim phenotype. Interestingly, the latter neutrophil subset is known to suppress lymphocyte proliferation ex vivo, but the underlying mechanism is largely unknown. We performed transcriptome analysis on the different neutrophil subsets to identify changes that are relevant for their functions. Neutrophil subsets were isolated by FACS sorting from the blood of healthy volunteers who were administered a single dose of lipopolysaccharide (LPS). The transcriptome was determined by microarray. The mobilized neutrophil subsets were characterized by specific transcriptome profiles reflecting their phase in neutrophil lifespan. Interestingly, the CD16bright/CD62Ldim suppressive neutrophils showed an interferon-induced transcriptome profile. This was confirmed by stimulation of peripheral neutrophils with IFNgamma. These cells acquired the capacity to suppress lymphocyte proliferation through the expression of programmed death ligand 1 (PD-L1). These data demonstrate that the suppressive phenotype of the neutrophil subset is induced by IFNgamma. Specific stimulation of neutrophils might have a pivotal role in regulating lymphocyte-mediated inflammation and autoimmune disease.
2013-05-01 | GSE42358 | GEO
Project description:Although the accumulation of neutrophils in the lungs and airways is common to many inflammatory lung diseases, including acute lung injury, the alterations that neutrophils undergo as they leave the peripheral circulation and migrate into the lungs have not been well characterized. Human volunteers were exposed to endotoxin by bronchoscopic instillation. The resulting air space neutrophil accumulation and peripheral blood neutrophils were isolated 16 h later, compared with circulating neutrophils isolated before or after to the pulmonary endotoxin exposure, and compared with circulating neutrophils exposed to endotoxin in vitro. Microarray analysis was performed on air space, circulatory, and in vitro endotoxin-stimulated neutrophils. Functional analysis included the determination of neutrophil apoptosis, chemotaxis, release of cytokines and growth factors, and superoxide anion release. Dramatic gene expression differences were apparent between air space and circulating neutrophils: approximately 15% of expressed genes have altered expression levels, including broad increases in inflammatory- and chemotaxis-related genes, as well as antiapoptotic and IKK-activating pathways. Functional analysis of air space compared with circulating neutrophils showed increased superoxide release, diminished apoptosis, decreased IL-8-induced chemotaxis, and a pattern of IL-8, macrophage inflammatory protein-1beta, monocyte chemoattractant protein-1, and tumor necrosis factor-alpha release different from either unstimulated or LPS-stimulated circulating neutrophils. Many of these changes are not elicited by in vitro treatment with endotoxin. Limited differences were detected between circulating neutrophils isolated before and 16 h after pulmonary endotoxin instillation. These results suggest that neutrophils sequestered in the lung become fundamentally different from those resident in the circulation, and this difference is distinct from in vitro activation with endotoxin.
2007-10-02 | E-GEOD-2322 | biostudies-arrayexpress
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.
2014-07-11 | E-GEOD-58920 | biostudies-arrayexpress
Project description:Neutrophils, a major innate immune cell population, are the most abundant circulating white blood cell in humans. They play a crucial role in host defense against infection; however, aberrant neutrophil activation may induce tissue damage via sterile inflammation. Neutrophil accumulation has been identified as a feature of the inflammatory response observed MASH and has been associated with liver fibrosis and cirrhosis. In the present study, we used RNA-Seq analysis of purified neutrophils to characterize their dysregulation in MASH.
2024-05-15 | GSE247467 | GEO
Project description:Neutrophils rapidly respond to inflammation and infection, but to which degree their functional trajectories after mobilization from the bone marrow can be shaped within the circulation remains vague. Phenotypic changes of circulating neutrophils caused by systemic inflammation are thought to result from several factors, including a “left shift” of the neutrophil compartment towards younger bone marrow-derived subsets. However, experimental limitations have so far hampered neutrophil research in human disease. Here, using innovative fixation and single-cell-based toolsets, we profile the human and murine neutrophil transcriptome and proteome during steady state and bacterial infection. We find that peripheral priming of circulating neutrophils leads to plastic shifts dominated by conserved upregulation of antimicrobial genes across neutrophil substates, facilitating pathogen containment. We show the TLR4/NF-κB-signaling dependent upregulation of canonical neutrophil activation markers like CD177/NB-1 during acute inflammation, resulting in functional shifts in vivo. Importantly, blocking de novo RNA synthesis in circulating neutrophils abrogates these plastic shifts and prevents the adaptation of antibacterial neutrophil programs by upregulation of distinct effector molecules upon infection. In summary, these data underline transcriptional plasticity as a relevant mechanism of functional neutrophil reprogramming during acute infection to foster bacterial containment within the circulation. This adds to the understanding of competing modes of adaptation to inflammatory challenges by the neutrophil compartment.
2024-05-21 | PXD048335 | Pride
Project description:Polymorphonuclear cells (neutrophils) play an important role in the systemic inflammatory response syndrome and the development of sepsis. These cells are essential for the defense against microorganisms, but may also cause tissue damage. Therefore, neutrophil numbers and activity are considered to be tightly regulated. Previous studies have investigated gene transcription during experimental endotoxemia in whole blood and peripheral blood mononuclear cells. However, the gene transcription response of the circulating pool of neutrophils to systemic inflammatory stimulation in vivo is currently unclear. We examined neutrophil gene transcription kinetics in healthy human subjects (n=4) administered a single dose of endotoxin (LPS, 2 ng/kg iv). In addition, freshly isolated neutrophils were stimulated ex vivo with LPS, TNFM-NM-1, G-CSF and GM-CSF to identify stimulus-specific gene transcription responses. Whole transcriptome microarray analysis of circulating neutrophils at 2, 4 and 6 hours after LPS infusion revealed activation of inflammatory networks which are involved in signaling of TNFM-NM-1 and IL-1M-NM-1 and IL-1M-NM-2. The transcriptome profile of inflammatory activated neutrophils in vivo reflects extended survival and regulation of inflammatory responses. We show that these changes in neutrophil transcriptome are most likely due to a combination of early activation of circulating neutrophils by TNFM-NM-1 and G-CSF and a mobilization of young neutrophils from the bone marrow. After LPS infusion blood was taken at t=0, t=2, t=4 and t=6 hours. Neutrophils were isolated and gene expression of these cells was assessed. T=2, t=4 and t=6 were related to t=0 as control condition
2012-06-04 | E-GEOD-35590 | biostudies-arrayexpress
Project description:Sterile tissue injury after stroke causes lymphocyte contraction in lymphoid tissues and may decrease circulating IgA-levels. Intestinal Peyer’s patches (PP) harbor large numbers of IgA+ B cell precursors and plasma cells. Whether and how tissue injury triggers PP-B cell death, thereby mediating IgA-loss, is unknown. We found decreased circulating IgA levels in stroke and myocardial infarction patients. Experimental stroke and myocardial infarction in mice phenocopied the human situation. Decreased plasma and fecal IgA were accompanied by rapid and macroscopic shrinkage of PP caused by substantial losses of PP-resident IgA+ precursors and plasma cells in mice. Tissue injury induced neutrophil activation endowed with the release of toxic neutrophil extracellular traps (NETs). Antibody-mediated or genetically- induced neutrophil loss, digestion of NETs, or inhibition of their release by the Gasdermin D blockade completely prevented lymphocyte loss and PP shrinkage. We also identified NETs in the plasma of stroke and myocardial infarction patients. Hence, tissue injury induces systemic NET-release, which might be targeted to maintain immune homeostasis at mucosal barriers.
2024-02-28 | PXD044644 | Pride
Project description:Low density neutrophils (LDN) co-purify with mononuclear cells during density-gradient separation of peripheral blood. LDN are rarely detected in healthy individuals but appear in the peripheral blood (PB) of patients with inflammatory diseases, malignancies and in individuals treated with granulocyte colony-stimulating factor (G-CSF). Using human G-CSF mobilized LDN, this study reveals that LDN are transcriptionally, proteomically and metabolically distinct from other neutrophils. Single Cell RNAseq analysis reveals that LDN from these donors comprise one major population, with multiple interrelated transcriptional states, representing the continuum of neutrophil maturation. LDN can engage both glycolysis and mitochondrial respiration to meet energy demands, which affords increased metabolically flexibility in the nutrient perturbed tissue environments found in tumours and inflammatory diseases. This study also identifies upregulated expression of CD98, a transporter for branch chain and aromatic amino acids, as a cell surface marker for LDN in peripheral blood. CD98 may contribute to the functional properties of LDNs and represents a useful phenotypic marker.
2022-12-19 | PXD023952 | Pride
Project description:Recent studies using advanced techniques such as single cell RNA sequencing (scRNAseq), high parameter flow cytometry, and proteomics have revealed that neutrophils are more heterogeneous than previously thought. Unique subsets have been identified in the context of bacterial and parasitic infections, cancer and tissue injury and repair. The diversity of subsets that predominate the neutrophil infiltrate in each of these conditions appears dependent on the type of injury and timing of neutrophil involvement in the disease state. We previously described a subpopulation of immature Ly6Glow neutrophils that accumulate in the peritoneal cavity 3 days following intraperitoneal (i.p.) administration of the fungal cell wall extract, zymosan. In contrast, conventional, mature Ly6Ghi neutrophils dominate the peritoneal infiltrate 4 hours following i.p. zymosan injection. The 3 day Ly6Glow, but not 4 hour Ly6Ghi, neutrophils induce neurite outgrowth of explanted neurons in vitro. Furthermore, they drive axon regeneration in the optic nerve (ON) and spinal cord (SC) when adoptively transferred in proximity to the eye after ON crush injury or SC dorsal column transection, respectively. These pro-regenerative effects are mediated in part, by the production of the growth factors insulin-like growth factor-1 (IGF-1) and nerve growth factor (NGF). In the current study, we expand upon on our previous observations by performing an in-depth characterization of the zymosan-modulated neutrophil subsets through scRNAseq and high parameter flow cytometry. Circulating neutrophils, harvested from unmanipulated mice, were analyzed in parallel as a control. We found that each of the unfractionated neutrophil populations is diverse, composed of multiple distinct clusters. Zymosanmodulated 3-day Ly6Glow neutrophils contain a cluster that expresses neuroprotective growth factors in association with markers of alternative activation. Furthermore, the unfractionated neutrophil populations were analyzed by liquid chromatography/ mass spectrometry for proteomic analysis. This new proteomic data identified a wide variety of proteins and pathways that contribute to the alternative activation state of these pro-regenerative 3 day Ly6Glow neutrophils. Collectively, the data generated by the current study constitute another step towards pinpointing the identity and mechanisms of action of pro-regenerative, neuroprotective neutrophil subsets.
2022-06-01 | GSE200445 | GEO