Project description:Purpose: Identification of neutrophils at different stages of maturation and their phenotypic and functional differences remain unknown. The goal of this study was to confirm maturation status of Ly6GIntermediate, Ly6GHigh and Ly6G+ neutrophils, the consistency of this means of identication in disease, and identify functional differences. Methods: Ly6G+, Ly6GIntermediate and Ly6GHigh neutrophil populations were isolated by FACS into individual cell populations from PBS controls, acute-LPS inflammation and pancreatic adenocarcinoma (PDAC) bearing mouse tissues (metastatic KPC, non-metastatic KPflC and WT controls). RNA-Sequencing was performed on bulk isolated populations Results: We identified Ly6GIntermediate as bona-fide immature neutrophils and analysis of Ly6G expression level as a consistently accurate marker of identifying neutrophil maturity. Gene expression was influenced by maturation stage, tissue of isolation, developmental niche and pathology. Function associated expression differences were maintained between Ly6GIntermediate and Ly6GHigh neutrophils throughout tissues and pathology.
Project description:This dataset combines transcriptional and chromatin analysis of neutrophils from the air pouch mouse model, an in vivo model of acute inflammation, with transcriptional analysis of Hoxb8 neutrophils, an in vitro model of neutrophil development.
Project description:Neutrophil recruitment is pivotal to host defense against microbial infection, but also contributes to the immunopathology of disease. We investigated the mechanism of neutrophil recruitment in human infectious disease by bioinformatic pathways analysis of the gene expression profiles in the skin lesions of leprosy. In erythema nodosum leprosum (ENL), which occurs in patients with lepromatous leprosy (L-lep), and is characterized by neutrophil infiltration in lesions, the most overrepresented biologic functional group was “cell movement” including E-selectin, which was coordinately regulated with IL-1. In vitro activation of TLR2, upregulated in ENL lesions, triggered induction of IL-1, which together with IFN-, induced E-selectin expression on, and neutrophil adhesion to endothelial cells. Thalidomide, an effective treatment for ENL, inhibited this neutrophil recruitment pathway. The gene expression profile of ENL lesions comprised an integrated pathway of TLR2/FcR activation, neutrophil migration and inflammation, providing insight into mechanisms of neutrophil recruitment in human infectious disease. 6 ENL skin lesions and 7 Lepromatous leprosy skin lesions
Project description:The purpose of the present study was to elucidate the exocytic events in neutrophil-mediated inflammation. Therefore, we first investigated the expression of genes implicated in these exocytic events in a cell model for neutrophils, the promyelocytic HL-60 cell line. By the addition of 1.3% DMSO during 4.5 days, these cells are differentiated into neutrophil-like dHL-60 cells. Non-differentiated cells served as control. Upon cell differentiation, we i) identified differentially expressed genes implicated in the mechanisms of vesicle transport, and ii) found some of them being upregulated. This upregulated gene expression upon DMSO-maturation points towards a functional role in these cells. Secondly, we confirmed the expression of the selected genes in human neutrophils, isolated from venous blood of 4 different donors.
Project description:Generalized pustular psoriasis (GPP) is a severe disease featured by neutrophilic pustules and enhanced IL-36 inflammatory pathway in skin. Recently, a transcriptomic analysis of PBMCs and neutrophils in MPO-deficient GPP patients has been performed in stable disease state. However, transcriptomic profiling of PBMCs during acute flare of GPP is unclear. Here, we reported a predominant neutrophil signature in GPP PBMC and a marked increase in the CD66+CD16+ low-density neutrophils (LDNs) within the PBMC fraction of acute GPP patients. Transcriptomic and functional analysis of LDNs revealed a hypoinflammatory phenotype yet enhanced release of neutrophil granule proteases, implicating that LDNs might contribute to the IL-36-mediated inflammation in GPP patients.
Project description:Neutrophil recruitment is pivotal to host defense against microbial infection, but also contributes to the immunopathology of disease. We investigated the mechanism of neutrophil recruitment in human infectious disease by bioinformatic pathways analysis of the gene expression profiles in the skin lesions of leprosy. In erythema nodosum leprosum (ENL), which occurs in patients with lepromatous leprosy (L-lep), and is characterized by neutrophil infiltration in lesions, the most overrepresented biologic functional group was 'cell movement' including E-selectin, which was coordinately regulated with IL-1beta. In vitro activation of TLR2, upregulated in ENL lesions, triggered induction of IL-1beta, which together with IFN-gamma, induced E-selectin expression on, and neutrophil adhesion to endothelial cells. Thalidomide, an effective treatment for ENL, inhibited this neutrophil recruitment pathway. The gene expression profile of ENL lesions comprised an integrated pathway of TLR2/FcR activation, neutrophil migration and inflammation, providing insight into mechanisms of neutrophil recruitment in human infectious disease.
Project description:BPIFA1 (BPI-fold containing group A member 1) is an airway host-protective protein with immunomodulatory properties that binds to LPS and is regulated by infectious and inflammatory signals. Differential expression of BPIFA1 has been widely reported in lung disease, yet the biological significance of this observation is unclear. We sought to understand the role of BPIFA1 fluctuations in modulating lung inflammation. We treated WT and bpifa1-/- mice with intranasal LPS and performed transcriptomic analyses to determine the immune effects of BPIFA1 deficiency. Transcriptomic analysis revealed a signature of 379 genes that differentiated bpifa1-/- mice from WT. During acute lung inflammation, the most strongly downregulated innate immunity genes in bpifa1-/- mice were cxcl9 and cxcl10. This was consistent with significantly decreased expression of IFN-g, IFN-l and downstream IFN-stimulated genes (ISG) and IFN-regulatory factors (IRF) important for innate immune responses.