Naive CD4+ T cell expansion defined by CD25 and CD31 expression
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ABSTRACT: Naive CD4 T cells defined as (TCRab+ CD4+ CD45RA+ CD62L+ CD127+) were further divided by CD25 and CD31 expression with FACS-based flow sorting. Subsequently, gene expression levels of each subset were assessed by using transcription arrays.
Project description:CD20+ B cells were analyzed ex-vivo using to determine technical and biological gene expression variability, in order to design well-powered experiments.
Project description:Single cell transcriptomic analysis of human CD25+ CD127- CD4+ Treg cells and CD25- CD127+ CD4+ Tconv cells isolated from peripheral blood from two different donors
Project description:We used single cell RNA sequencing (scRNA-seq) to analyze the difference among CD25+CD127-Treg, iTreg, and iCD4+Treg-like. We expanded CD25+CD127-Treg by TCR stimulation in the presense of AS2863619, anti-TNFR2 antibody, and TGFbeta (AMT). We differentiated iTreg from naive CD4+T cells and iCD4+Treg-like from iPSC-derived CD4+T cells in the presence of TCR stimulation, AS2863619, anti-TNFR2 antibody, TGFbeta, and rapamycin (AMRT).
Project description:Transcriptome sequencing of sorted CD127+ DN T cells, MAIT, NKT, and CD127+CD4 and Cbir DN to identify transcriptional signatures of CD4 and CD8 double negative T cells.
Project description:We compared gene expression among central memory (CD62L+CD127+; pop1), effector memory (CD62L-CD127+; pop2), and terminal effector (CD62L-CD127-; pop4) isolated from a single-cell-derived WT1-specific CD4+ T cell clone.
Project description:Teff cells (CD4+CD25-CD127+CD45RO+) were co-cultured with and without sorted Treg subsets plus dendritic cells for 18h. All the sorted Teff cells are gated on CD4+CD25-CD127(hi), and further sorted for CD45RO- and CD45RO+ phenotype.
Project description:All the sorted Treg cells are gated on CD4+CD25+CD127(low), followed by gating on various subsets using markers indicated in the sample names. All the sorted Teff cells are gated on CD4+CD25-CD127(hi), and further sorted for CD45RO- and CD45RO+ phenotype.
Project description:Comparison of gene expression profile of CD4+ CD25+, CD4+ CD25- CD45RBlow LAG3+ and CD4+ CD25- CD45RBlow LAG3- T cells. Naive CD4+CD25-CD45RBhigh T cells were used as a reference for pair comparison with values from the three other subsets.
Project description:Increased T cell IL-7Rα (CD127) signaling is associated with poor prognosis in ANCA-associated vasculitis patients, whereas genes linked to T cell exhaustion such as PD-1 correlate with fewer relapses and better patient outcomes. This study characterised intrarenal CD127 and PD-1 expressing CD4+ and CD8+ T cells in mice with anti-MPO glomerulonephritis by RNA sequencing and examined the functional role of IL-7Rα in experimental glomerulonephritis mediated by anti-MPO T cell autoimmunity. There were 3,738 and 2,726 genes differentially expressed between intrarenal CD127-PD-1+ and CD127+PD-1- CD8+ and CD4+ T cells, respectively, with a substantial overlap of differentially expressed genes between CD8+ and CD4+ T cells. Both CD127-PD-1+ CD8+ and CD4+ T cells were enriched for previously described T cell exhaustion signatures that predict prognosis in autoimmune disease. As effector memory T cells drive inflammation, we blocked CD127 after the induction of anti-MPO autoimmunity. Compared with control IgG, anti-IL-7Rα antibodies limited histological injury, reduced albuminuria and reduced numbers of glomerular and interstitial leukocytes, with reduced intrarenal chemokine and pro-inflammatory cytokine expression. Intrarenal effector memory and exhausted CD4+ and CD8+ T cells are present in experimental anti-MPO glomerulonephritis. Neutralising effector memory T cells via the IL-7Rα after the induction of autoimmunity limits intrarenal inflammation and disease. IL-7Rα may be a therapeutic target in ANCA-associated vasculitis.