Project description:Anergic T cells are unable to respond to antigen though their TCR is specific for it. Chronic infections and cancer can induce a dsyfunctional immune response which cannot clear the cause of disease. We asked in how far anergic T cells are similar to antigen-exhausted ones.
Project description:The NR4A family of nuclear receptors are upregulated by acute and chronic antigen stimulation, and play redundant, tolerogenic roles in T and B cells. To bypass their obligate function in the Treg lineage, we generated competitive chimeras with Nr4a1-/-Nr4a3-/- (double knockout or DKO) bone marrow mixed with congenically marked WT bone marrow at a 1:5 ratio. As a result of a defect in Treg production by DKO cells, Treg in mixed chimeras are almost exclusively of WT origin. By contrast, we generate an approximately 1:1 DKO:WT ratio in the peripheral T cell compartment because DKO cells escape negative selection. After reconstitution, we observed pronounced accumulation of anergic DKO CD4 T cells, but a defect in functional tolerance resulting in autoantibody production. Based on these studies and published literature, we hypothesize that the NR4A family regulate TCR-induced transcripts in both naive and anergic CD4 T cells. To identify such targets, we sorted naive and anergic CD4 T cells of either WT or DKO genotype (CD45.1+ or CD45.1-) on the basis of CD73/FR4/CD44 expression from these chimeras directly into RLT buffer. In parallel, we stimulated sorted naive CD4 T cells of each genotype in vitro with plate-bound anti-CD3 and anti-CD28 stimulation for 3 hrs and then generated RLT lysates. We made biological triplicate samples of each condition (naive WT and DKO, naive stimulated WT and DKO, anergic WT and DKO) resulting in 6 different populations x 3 replicates = 18 samples. Samples were subjected to library prep and bulk RNA sequencing to identify DEG.
Project description:Up to now the role of tumor-specific pTregs and anergic cells during tumor development is not fully understood. Here we used a genetically-induced tumor expressing a MHC-II restricted DBY model antigen to characterize the tumor-induced pTregs and anergic cells that arise early during tumor development.
Project description:Array of global miRNA expression during an in vivo effector and anergic T cell response 250 ng RNA isolated from mouse D011.10+ CD4 T cells, activated in vivo for 5 days
Project description:compare gene expression profiles between normal and anergic T cells and identify upregulated genes in anergic T cells Experiment Overall Design: RNA from normal Th1 T cell clone and anergic Th1 T cell clone made anergic by plate-bound anti-CD3 antibody were isolated and amplified for microarray analysis
Project description:We aim to find the gene-specific effects of Rnmt KO in mouse CD4 T cells. TMT proteomics datasets were generated to find out which proteins are dependent on RNMT for their expression.
Project description:We aim to find the gene-specific effects of Rnmt KO in mouse CD4 T cells. TMT proteomics datasets were generated to find out which proteins are dependent on RNMT for their expression.