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    Home > Active Ingredient News > Immunology News > The researchers published a map of single-cell development and heterogeneity in the stable and inflammatory state of neutral granulocytes.

    The researchers published a map of single-cell development and heterogeneity in the stable and inflammatory state of neutral granulocytes.

    • Last Update: 2020-09-11
    • Source: Internet
    • Author: User
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    On July 27, 2020, the Li Cheng Research Group of the School of Life Sciences of Peking University, in collaboration with the Luo Hongbo Research Group of Harvard Medical School and the Ma Fengxia Research Group of the Institute of Hematology of the Chinese Academy of Medical Sciences, published a research paper entitled "Single-cell transcriptome profiling reveals neutrophil hetereity homeostasis and application" in the journal Nature Immunology.
    neutral granulocytes are the main component of the natural immune system, as the largest number of circulating white blood cells in the cell type, is the host against invasive bacteria and other pathogens the first line of defense.
    However, at present, the sub-groups of neutral granulocytes mainly rely on classical morphology, the heterogeneity of neutral granulocytes has been controversial, there is no unified understanding, this difference hinders the development of neutral granulocyte research.
    the number and level of gene expression in neutral granulocytes is lower than in other blood and immune cell types, and it is more difficult to capture RNA technically.
    , since the half-life of neutral granulocytes is only 15-20 hours, the details of the operation of the experiment are also higher requirements.
    the authors performed neutral granulocyte separation and single-cell RNA sequencing on bone marrow, perennial blood and spleen in mice by optimizing experimental conditions, and established a neutral granulocyte map under steady state conditions in combination with bio-informational methods.
    in order of developmental maturity, the neutral granulocytes in the bone marrow can be divided into 5 groups, including 3 sub-groups (G0, G1, G2) at the stage of division and 2 more mature sub-groups (G3, G4) after division.
    the authors analyzed the characteristics of each group in detail, and verified the existence of these groups by combining flow cell markers.
    study found that although the heterogeneous cell population defined by single-cell RNA sequencing has a high correlation with the classical morphological credit group as a whole, the single-cell sequencing technique can identify the same morphologically identical G3 and G4 granulocyte populations, which have differences in transcription groups, and increase the understanding of heterogeneity of mature granulocytes in bone marrow.
    Based on the heterogeneity of particle protein genes expressed between subpopulations, the authors proposed the "sorting hypothesis" of particle protein production and identified new transcription factors specifically raised during the development of neutral granulocytes through transcription control network analysis.
    the authors then analyzed the neutral granulocytes in the outer blood and spleen, and found that there were three groups of mature neutral granulocytes, PMNa, PMNb, and PMNc, which had different transcriptional group characteristics.
    RNA velocity analysis revealed that these mature groups have different bone marrow sources and complex inter-group transformational relationships: 1) PMNa and PMNb are mainly developed by G3 and G4 of bone marrow, respectively;
    Using immunofluorescence imaging techniques, the authors further validated the unique spatial distribution of mature neutral granulocytes (PMNbs) in the spleen of genes associated with high expression interferon stimulation, suggesting that the sub-group may have host defense functions that are different from those of other sub-groups.
    In addition, the authors also mapped heterogeneous populations defined by single-cell sequencing in previous literature to the neutral granulocyte reference map established in this study, unifying the classification of heterogeneity of neutral granulocytes in mice in a stable state.
    Finally, to study the effects of bacterial infection on neutral granulocyte development and sub-groups, the authors isolated the bone marrow, extrinsic blood, spleen and neutral granulocytes of acute perititis mice for single-cell RNA sequencing and analysis.
    found no change in the heterogeneity of neutral granulocytes in the inflammatory state, but significant changes were made in the proportion of populations, functional characteristics, transcription factor expression and group transformation pathways.
    analysis and experimental results show that acute myelin hematosis is done mainly by enhancing early neutral granulocytosis activity and shortening the ripening time of neutral granulocytes after division, and these findings provide potential molecular targets for the treatment of acute bacterial infections.
    Overall, the study systematically depicts for the first time the dynamic changes of heterogeneous groups and transcriptomes in the process of heterogeneous groups and transcriptional groups in the process of heterogeneous cells maturation, differentiation and destiny decision-making in a steady and inflammatory state from a single-cell scale, integrating existing classifications of nexual granulocyte heterogeneity in the field, and providing valuable reference data resources for further study of sexual granulocytes () 
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