Project description:Primary RNASeq data for progenitor, resident, and stimulated (C.alb, LPS, injury, APAP+ starved overnight and pIC) mononuclear phagocytes from fourteen organs.
Project description:The mononuclear phagocytic system (MPS) is characterized by functional diversity and plasticity, including within the tumor microenvironment (TME). Tumor-associated mononuclear phagocytes (TMPs) within the TME come from various ontological origins. To determine how Fcmr deficiency might impact the functional heterogeneity of mature mononuclear phagocytes (MPs) within the TME, we performed single-cell RNA sequencing (scRNAseq) on MPs isolated from B16 tumors resected from Fcmr-/- and Fcmr+/+ mice. To evaluate a homogenous population of MPs within the TME we chose to examine cells harboring expression of markers found in more mature, differentiated MPs. Therefore, we then sorted MerTk+ CD64+ TMPs by FACS and subjected these cells to single-cell transcriptomic analyses using the Chromium 10X Genomics platform. We obtained RNAseq profiles from 6352 Fcmr+/+ and 7999 Fcmr-/- TMPs.
Project description:The small intestine contains lymph node-like structures termed Peyer’s patches (PP). In adult mice they contain active germinal centers reminiscent of active crosstalk with microbiota. In neonatal mice, however, homeostatic immune activation does not take place. We wanted to test whether immune maturation in neonatal mice is stalled due to a reduced presence or function of antigen presenting mononuclear phagocytes (MNP). Thus we subjected PP MNP of postnatal day (PND) 11 and adult mice to scRNAseq in steady state or after immune activation with the TLR7 agonist R848 that induces type I interferon and TNFa after oral gavage. Our results show an altered subset distribution of MNP in neonatal mice (predominance of cDC1 and cDC2b, reduced proportion of cDC2a) and a reduced potential to take up microorganisms and process/present antigen as well as decreased type I interferon mediated gene expresseion. R848 administration could partly overcome this maturation defect of neonatal MNP by increasing the proportion of CCR7+ mature cDC as well as upregulation of interferon I dependent genes.
Project description:Muscle injury triggers inflammation in which infiltrating mononuclear phagocytes are crucial for tissue regeneration. The interaction of the CCL2/CCR2 and CX3CL1/CX3CR1 chemokine axis that guides phagocyte infiltration is incompletely understood. Here, we show that CX3CR1 deficiency promotes muscle repair and rescues Ccl2-/- mice from impaired muscle regeneration as a result of altered macrophage function, not infiltration. Transcriptomic analysis of muscle mononuclear phagocytes reveals that Apolipoprotein E (ApoE) is up-regulated in mice with efficient regeneration. ApoE treatment enhances phagocytosis by mononuclear phagocytes in vitro, and restores phagocytic activity and muscle regeneration in Ccl2-/- mice. Because CX3CR1 deficiency may compensate for defective CCL2-dependant monocyte recruitment by modulating ApoE-dependent macrophage phagocytic activity, targeting CX3CR1 expressed by macrophages might be a powerful therapeutic approach to improve muscle regeneration.
Project description:This study aimed to investigate the protective mechanisms of the Traditional Chinese Medicine Tongfu Pingchuan decoction (TFP) in the cecum of septic rats. Thirty rats were randomly divided into three groups: Sham, Model (sepsis induced by cecal ligation and puncture), and TFP (sepsis treated with TFP). Rats received 1 mL/kg TFP or PBS via gavage twice daily for one week, after which whole blood and cecum tissues were collected. Transcriptome sequencing was conducted on six cecum tissues per group. Bioinformatics analyses predicted the potential mechanisms of TFP, which were subsequently validated in cecum tissues and RAW264.7 cell cultures. We found that Pathways such as TNF signaling, complement and coagulation cascades, fatty acid metabolism, and PPAR signaling were activated, while cholinergic synapse, ECM-receptor interaction, and VSMC contraction were inhibited in the Model group compared to Sham. These changes were largely reversed in the TFP group. Chrm4 and Pygm were identified as potential TFP targets, as they were overlapping differentially expressed genes (DEGs) in the predicted TFP targets and an identified WGCNA module of interest. Chrm4 was downregulated in the Model group and restored by TFP treatment. In the Model group, M1 macrophages were induced, while M2 macrophages were suppressed. TFP inhibited M1 macrophages and further enhanced M2 macrophages. These effects were corroborated in LPS-stimulated RAW264.7 cells, where TFP also modulated cytokine levels (IL-6, IL-10, TNF-α). The effects of TFP were negated by Boc-2 (a PCTR1 inhibitor) or Chrm4 silencing. Additionally, TFP suppressed IL1RN, p-ERK2/ERK2, and p-AKT/AKT levels in LPS-treated RAW264.7 cells, but these effects were abolished upon Chrm4 silencing. Collectively, we show that TFP alleviates sepsis by promoting Chrm4/IL1RN-regulated macrophage polarization and facilitating inflammation resolution.
Project description:Aim of this project was to examine the global gene expression profiles of mononuclear phagocytes recruited from peripheral blood to the alveolar space following alveolar deposition of the TLR2-ligand Pam3CSK4 in transgenic CX3CR1+/GFP mice.