Project description:MED23, a subunit of the Mediator coactivator complex, is important for the expression of a subset of MAPK/ERK pathway-dependent target genes; however, the genes in this subset varies between cell types. MAPK/ERK pathway-dependent processes are essential for T-cell development and function, but whether MED23 has a role in this context is unknown. We generated Med23 conditional knockout mice and induced Med23 deletion in early T cell development using the lineage specific Lck-Cre transgene. While the total cell number and distribution of cell populations in the thymuses of Med23flox/flox;Lck-Cre mice were essentially normal, MED23 null T-cells failed to efficiently populate the peripheral lymphoid organs. MED23 null thymocytes displayed decreased expression of the MAPK/ERK-responsive genes Egr1, Egr2, as well as of the membrane glycoprotein Cd52 (CAMPATH-1). MED23 null CD4 single-positive thymocytes also showed decreased expression of KLF2 (LKLF), a T cell master regulatory transcription factor. Indeed, similarities between the phenotypes of mice lacking MED23 or KLF2 in T-cells suggest that KLF2 deficiency in MED23 null T-cells is one of their key defects. Mechanistic experiments using MED23 null MEFs further suggest that MED23 is required for full activity of the MAPK-responsive transcription factor MEF2, which has previously been shown to mediate Klf2 expression. In summary, our data indicate that MED23 has critical roles in enabling T-cells to populate the peripheral lymphoid organs, possibly by potentiating MEF2-dependent expression of the T-cell transcription factor KLF2. 12 samples, 2 of each genotype (Lck-Cre, Med23flox/flox and Med23flox/flox;Lck-Cre) both with mock and anti-CD3 treatment
Project description:tPTEN-/- mice display a deletion of the PTEN tumor suppressor gene specifically in T cells (cross PTEN flox/flox x lck-Cre). They develop T cell lymphoma with a primary thymic tumor and invasion of most organ at late stage of the disease. Gene profiling was performed on whole tumors (ST3 stage 3 invasive lymphoma) and normal thymocytes (wt) using Affymetrix-MoGene-1_0-st-v1 chips.
Project description:Human CD4+ T cells were left untreated or stimulated with 5 μg/ml plate-bound anti-CD3 antibody (clone OKT3, 14-0037-82, eBioscience) and 10 μg/ml soluble anti-CD28 antibody (clone CD28.2, 14-0289-82, eBioscience) for 6 h in RPMI1640 medium supplemented with 10% FBS, 1% L-glutamine, and 1% penicillin-streptomycin.
Project description:We report the differential mRNA expressino of WT and Chi3l1 KO Th1 cells. We cultured WT and Chi3l1 KO naive CD4 T cells under Th1 skewing condition : plate-bound anti-CD3/28 antibody (2ug/mL), IL-12 0.2ng/mL, IL-2 50U/mL, anti-IL-4 neutralizing antibody 2ug/mL, for 3 days. We found different Th1 regulatory and tumoricidal-related gene expression in Chi3l1 KO T cells.
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:Control (cMet+ T cells less than 0.5%) and cMet-enriched (7%) T cells were activated with plate-bound anti-CD3/CD28 and gene tracription was measured
Project description:tPTEN-/- mice display a deletion of the PTEN tumor suppressor gene specifically in T cells (cross PTEN flox/flox x lck-Cre). They develop T cell lymphoma with a primary thymic tumor and invasion of most organ at late stage of the disease.
Project description:Lamtor1-KO BMDCs isolated from Lamtor1flox/flox X CD11c-Cre mice were lysed by Lysis Buffer A and immunoprecipitated by anti-Lamtor1 antibody (D11H6).
