Project description:The generation of TCRαβ and TCRγδ T cells proceeds through distinct developmental stages in which changing regulatory events control differentiation and lineage outcome. To clarify the underlying mechanisms, we employed RNAseq, ATACseq and ChIPmentation on well-defined thymocyte subsets that represent the continuum of human T cell development. The chromatin accessibility dynamics uncover stage-specific regulatory mechanisms and reveal that human T-lineage commitment is marked by the GATA3- and BCL11B-dependent closing of PU.1 sites. While the ß-selection checkpoint is marked by transcriptional changes and a temporary increase in H3K27me3 without modifications in open chromatin, emerging γδ T cells, that originate from common precursors as ß-selected cells, show dramatic changes in chromatin accessibility which results from strong TCR signaling. Furthermore, we unravel distinct chromatin landscapes between CD4 and CD8 αβ T cells that support their effector functions and reveal gene-specific regulatory mechanism that define mature T cells. This resource provides an important framework for studying gene regulatory mechanisms that drive human normal and malignant T cell development.
Project description:The generation of TCRαβ and TCRγδ T cells proceeds through distinct developmental stages in which changing regulatory events control differentiation and lineage outcome. To clarify the underlying mechanisms, we employed RNAseq, ATACseq and ChIPmentation on well-defined thymocyte subsets that represent the continuum of human T cell development. The chromatin accessibility dynamics uncover stage-specific regulatory mechanisms and reveal that human T-lineage commitment is marked by the GATA3- and BCL11B-dependent closing of PU.1 sites. While the ß-selection checkpoint is marked by transcriptional changes and a temporary increase in H3K27me3 without modifications in open chromatin, emerging γδ T cells, that originate from common precursors as ß-selected cells, show dramatic changes in chromatin accessibility which results from strong TCR signaling. Furthermore, we unravel distinct chromatin landscapes between CD4 and CD8 αβ T cells that support their effector functions and reveal gene-specific regulatory mechanism that define mature T cells. This resource provides an important framework for studying gene regulatory mechanisms that drive human normal and malignant T cell development.
Project description:Background: Binding of the programmed death-1 (PD-1) receptor to its ligands (PD-L1/2) transduces inhibitory signals that promote exhaustion of activated T cells. Blockade of the PD 1 pathway is widely used for cancer treatment, yet the inhibitory signals transduced by PD-1 in T cells remain elusive. Methods: Expression profiles of human CD8+ T cells in resting, activated (CD3+CD28) and PD-1-stimulated cells (CD3+CD28+PD-L1-Fc) conditions were evaluated by RNA-seq. Bioinformatic analyses were used to identify signaling pathways differentially regulated in PD 1-stimulated cells.
Project description:Transcriptional profiling of Double Negative (CD4-/CD8-) T-cells isolated from SIV infected Sooty Mangebys. DN T-cells were stimulated through the T-Cell receptor using anti CD3/CD28 antibodies for 4 hours and compared to unstimulated DN T-cells.
Project description:isp-1;sod double mutants have decreased lifespan, increased resistance to oxidative stress and slow physiologic rates. We performed RNA sequencing to compare gene expression between isp-1 mutants and isp-1;sod-3 and isp-1;sod-5 double mutants
Project description:Transcriptional profiling of Double Negative (CD4-/CD8-) T-cells isolated from SIV infected Sooty Mangebys. DN T-cells were stimulated through the T-Cell receptor using anti CD3/CD28 antibodies for 4 hours and compared to unstimulated DN T-cells. Two condition experiment, Stimulated vs Unstimulated. 4 animals tested, each with Stim vs Unstim, a dye flip of Stim vs Unstim, Stim vs Stim and Unstim vs Unstim.
Project description:Genome wide expression profiles (RNAseq) of human CD8+ T cells in resting, activated (CD3+CD28) and PD-1-stimulated cells (CD3+CD28+PD-L1-Fc) at diferent time points
Project description:Epidemiologic studies demonstrate that women from cultures that consume high levels of dietary soy have reduced breast cancer rates compared to women from cultures where soy consumption is typically much lower. The types of soy products consumed can also differ with Asian cultures consuming primarily minimally refined soy products while Western cultures often consume more highly refined soy products such as isolated soy protein (ISP). Our previous work showed that lifetime exposure to a diet containing 20% ISP promoted mammary tumor development in MTB-IGFIR transgenic mice. In this study, lifetime exposure to lower levels of ISP were evaluated (5% ISP and 1% ISP) to determine whether more moderate levels of ISP could protect against mammary tumorigenesis. A standard rodent diet, Teklad 2018 was also included in this study and Teklad 2018 contains a less refined form of soy, namely soybean meal. MTB-IGFIR mice fed ISP diets, independent of the concentration, displayed increased mammary tumor incidence and reduced tumor latency compared to MTB-IGFIR mice fed a 20% casein diet. Unexpectedly, MTB-IGFIR mice fed Teklad 2018 were completely protected against mammary tumor development. Although RNA sequencing of mammary tumors from ISP or casein fed mice did not identified gene expression patterns associated with the ISP diets, the ISP diets consistently promoted the expression of contractile related proteins in pubertal mammary glands. Therefore, lifetime exposure to ISP may alter gene expression in pubertal mammary glands rendering them more susceptible to transformation. Based on these findings women may want to avoid highly refined soy products such as ISP and switch to less refined forms of dietary soy until additional studies can be performed.
Project description:Epidemiologic studies demonstrate that women from cultures that consume high levels of dietary soy have reduced breast cancer rates compared to women from cultures where soy consumption is typically much lower. The types of soy products consumed can also differ with Asian cultures consuming primarily minimally refined soy products while Western cultures often consume more highly refined soy products such as isolated soy protein (ISP). Our previous work showed that lifetime exposure to a diet containing 20% ISP promoted mammary tumor development in MTB-IGFIR transgenic mice. In this study, lifetime exposure to lower levels of ISP were evaluated (5% ISP and 1% ISP) to determine whether more moderate levels of ISP could protect against mammary tumorigenesis. A standard rodent diet, Teklad 2018 was also included in this study and Teklad 2018 contains a less refined form of soy, namely soybean meal. MTB-IGFIR mice fed ISP diets, independent of the concentration, displayed increased mammary tumor incidence and reduced tumor latency compared to MTB-IGFIR mice fed a 20% casein diet. Unexpectedly, MTB-IGFIR mice fed Teklad 2018 were completely protected against mammary tumor development. Although RNA sequencing of mammary tumors from ISP or casein fed mice did not identified gene expression patterns associated with the ISP diets, the ISP diets consistently promoted the expression of contractile related proteins in pubertal mammary glands. Therefore, lifetime exposure to ISP may alter gene expression in pubertal mammary glands rendering them more susceptible to transformation. Based on these findings women may want to avoid highly refined soy products such as ISP and switch to less refined forms of dietary soy until additional studies can be performed.