Project description:To identify cell composition and characterize transcriptional regulation in regulatory T (Treg) cells derived from mice subjected to soluble tachyzoite antigen (STAg) of Toxoplasma gondii treatment, we prepared splenic CD4+CD25+ Treg cells from PBS and STAg treated mice, and performed scRNA-seq using the 10× Genomics method.
Project description:MAP4K family kinases are key kinases for T-cell-mediated immune responses; however, in vivo roles of MAP4K2 in immune regulation remain unclear. Using T-cell-specific Map4k2 conditional knockout (T-Map4k2 cKO) mice, single-cell RNA sequencing (scRNA-seq), and mass spectrometry analysis, we found that MAP4K2 interacted with DDX39B, induced FOXP3 gene expression, and promoted Treg differentiation. Mechanistically, MAP4K2 directly phosphorylated the DEAD box protein DDX39B, leading to DDX39B nuclear translocation and subsequent Foxp3 RNA splicing. MAP4K2-induced FOXP3 mRNA levels were abolished in DDX39B knockout T cells. Furthermore, T-Map4k2 cKO mice displayed the reduction of Treg population and sustained inflammation during remission phase of EAE autoimmune disease model. Remarkably, the anti-PD-1 immunotherapeutic effect on pancreatic cancer was drastically improved in T-Map4k2 cKO mice, Treg-specific Map4k2-deficient mice, adaptively transferred mice, or MAP4K2-inhibitor-treated mice. Consistently, scRNA-seq analysis of human pancreatic patients showed increased MAP4K2 levels in infiltrating Treg cells. Collectively, MAP4K2 promotes Treg differentiation by inducing DDX39B nuclear translocation, leading to the attenuation of tumor immunity.
Project description:MAP4K family kinases are key kinases for T-cell-mediated immune responses; however, in vivo roles of MAP4K2 in immune regulation remain unclear. Using T- cell-specific Map4k2 conditional knockout (T-Map4k2 cKO) mice, single-cell RNA sequencing (scRNA-seq), and mass spectrometry analysis, we found that MAP4K2 interacted with DDX39B, induced FOXP3 gene expression, and promoted Treg differentiation. Mechanistically, MAP4K2 directly phosphorylated the DEAD box protein DDX39B, leading to DDX39B nuclear translocation and subsequent Foxp3 RNA splicing. MAP4K2-induced FOXP3 mRNA levels were abolished in DDX39B knockout T cells. Furthermore, T-Map4k2 cKO mice displayed the reduction of Treg population and sustained inflammation during remission phase of EAE autoimmune disease model. Remarkably, the anti-PD-1 immunotherapeutic effect on pancreatic cancer was drastically improved in T-Map4k2 cKO mice, Treg-specific Map4k2- deficient mice, or MAP4K2-inhibitor-treated mice. Consistently, scRNA-seq analysis of human pancreatic or lung cancer patients showed increased MAP4K2 levels in infiltrating Treg cells. Collectively, MAP4K2 promotes Treg differentiation by inducing DDX39B nuclear translocation, leading to the attenuation of tumor immunity.
Project description:To understand whether hematopoietic stem cells (HSCs) take part in emergency granulopoiesis, WT C57BL6 mice were treated intraperitoneally with 35 ug of LPS (lipopolysacharide) to induce emergency granulopoiesis or PBS control. Mice were sacrificed 4 hours after the treatment andHSCs (Lin-c-Kit+Sca-1+CD48-CD150+) were sorted and subjected to scRNA-seq.
Project description:Two experimental groups were set with six samples of spleen from control mice injected with PBS. Total splenocyte were isolated, part of which were sorted in order to prepare Treg-depleted spleen samples.
Project description:We investigated early effects of IL2 treatment on mouse Treg chromatin accessibility by performing scATAC-seq on splenic Tregs isolated from mice 2h after treatment with recombinant IL2 or PBS vehicle control.
Project description:We report gene expression of Treg cells isolated from injured muscle in IL-33 vs PBS treated mice. Male Foxp3-GFP C57BL/6 reporter (2 months old) mice were injured intramuscularly with cardiotoxin/rIL-33 (0.3 ug/muscle). Tregs were sorted directly into Trizol from injured muscle 4 days post-injury.
Project description:We report gene expression of Treg cells isolated from injured muscle in IL-33 vs PBS treated mice. Male Foxp3-GFP C57BL/6 reporter (2 months old) mice were injured intramuscularly with cardiotoxin/rIL-33 (0.3 ug/muscle). Tregs were sorted directly into Trizol from injured muscle 4 days post-injury. Gene expression profiling of muscle Tregs from IL-33 vs PBS injured mice.
Project description:We compared the transcriptomes of mesenchymal cells and immune cells in mice sensitized and challenged with house dust mite (HDM) extracts and those in control PBS-treated mice, using single cell RNA-seq (scRNA-seq). Data with mesenchymal cells and data with immune cells were obtained from separate experiments. For scRNA-seq of mesenchymal cells, each sample was pooled from cells from 3 mice and 2 different samples per group (for a total of 6 mice) were used. For scRNA-seq of immune cells, each sample was from cells of one individual mouse.