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Transcriptomic analysis of human CD4+, CD8+ and DP T cells

ABSTRACT: Analysis of the transcriptome of human CD4+, CD8+ and DP T cells. The objective is to define the origin of the DP in melanoma

ORGANISM(S): Homo sapiens

PROVIDER: GSE141465 | GEO | 2020/04/17

REPOSITORIES: GEO

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Mapping of H2A.Z nucleosomes in CD4/CD8 DP thymocytes

Project description:H2A.Z ChIP-Seq in CD4/CD8 DP thymocytes

2017-02-07 | GSE87529 | GEO

Transcriptomic analysis of human CD4+, CD8+ and DP T cells

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Gene expression of CD4+CD8+ Tfh cells and CD4+CD8- Tfh in tonsil.

Project description:To understand physiological characteristics of CD4+CD8+ Double-positive (DP)-Tfh cells, we continued to conduct comparative transcriptome analysis of tonsillar Tfh cell populations, including DP-Tfh cells and CD4+CD8- single-positive (SP)-Tfh cells as a control. Results showed that DP-Tfh cells preferentially expressed transcripts related to CTLs, such as CD8 (CD8A and CD8B), Eomes, and granzymes, suggesting a possible cytotoxic attribute of DP-Tfh cells. Th1-cell related signature genes were also presented in DP-Tfh cells and cytokines like interferon (IFN)-gamma and interleukin (IL)-10 were found to be highly presented in DP-Tfh cells rather than SP-Tfh cells. Of note, the expression profile of authentic Tfh-cell (i.e. SP-Tfh cell) related genes like IL-4, IL-21, Bcl6, and Pou2af1 seemed to be shared with DP-Tfh cells.

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Gene expression analysis of Ets1-/- CD4+ CD8+ thymocytes

Project description:We performed ChIP-Seq for hallmark TFs (Ets1, Runx1), histone modification marks (H3K4me1, H3K4me2, H3K4me3, H3K27me3, H3K36me3), total RNA Pol II, short RNA-Seq as well as nucleosome mapping mainly in murine Rag2 -/- thymocytes. We also performed ChIP-Seq for E47 as well as nucleosome mapping, gene expression microarray analysis in CD4+ CD8+ WT and Ets1-/- DP thymocytes. Overall, we find a key role for the transcription factor Ets1, contributing towards alpha beta T cell lineage commitment via differential transactivation of stage-specific genes orchestrated by dynamic, co-association -mediated chromatin remodeling, as well as transcription dependent generation of a specialized chromatin structure at the TCR beta locus. Genome-wide analysis via ChIP-Seq for Ets1, Runx1, total RNA Pol II binding, H3K4me1, H3K4me2, H3K4me3, H3K27me3, H3K36me3, short RNA-Seq, Mnase-Seq in murine Rag2 -/- thymocytes, ChIP-Seq for E47, Mnase-Seq and gene expression microarray analysis in DP thymocytes Gene expression analysis of Ets1-/- CD4+ CD8+ thymocytes

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A population of CD4+CD8+ double-positive T cells associated with risk of plasma leakage in dengue viral infection

Project description:According to the WHO 2009 classification, dengue with warning signs (D+W) is at the risk of developing severe form of dengue disease. One of the most important warning signs is plasma leakage, which is a serious complication associated with higher morbidity and mortality. We report that the frequency of CD4+CD8+ double-positive (DP) T cells is significantly increased in patients at risk of developing plasma leakage. Transcriptomic analysis demonstrated that CD4+CD8+ DP cells were distinct from CD4+ Single Positive (SP) T cells, but co-clustered with CD8+ SP cells indicating a largely similar transcriptional profile. 20 significant differentially expressed (DE) genes were identified between CD4+CD8+ DP and CD8+ SP cells. These genes encode OX40 and CCR4 (TNFRSF4 and CCR4), as well as other molecules associated with cell signaling on the cell surface (NT5E, MXRA8, PTPRK). While 5 significant DE genes were identified, the profile of gene expression in CD4+CD8+ DP cells was mostly similar between patients with and without warning signs of plasma leakage, implying a role of CD4+CD8+ DP cells in plasma leakage through a quantitative increase rather than functional alteration. The differentially expressed genes identified could have diagnostic utility, perhaps as PCR-based diagnostic markers.

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Project description:Purpose: To establish if and how mediastinal and adipose tissue and aortic arch CD4+CD8+TCRb- cells (DPs) would differ from thymic DPs, we sorted DPs from all three regions by flow cytometry and subjected the cells to a low-input deep transcriptional profiling assay. Methods: Thymi, mediastinal adipose tissue, and aortic arches of five female, 8-week-old C57BL/6J mice were collected and pooled. Aortic arches and mediastinal adipose tissue were digested as previously described (PMID: 14597735) and single cell suspensions were generated from all three tissues. Four pools were generated per tissue. 400 viable CD4+CD8b+TCRb- cells were sorted into 8ul low-input lysis buffer containing RNaseH inhibitor, dNTPs, and 0.1% Triton X-100 in replicates. Material of 200 cells was reverse transcribed and amplified. Illumina libraries were prepared and sequenced. Results: A total of 2147 genes were differentially regulated between the different DP populations. Extrathymic DPs had 858 genes upregulated and 1284 genes downregulated compared to thymic DPs. 94 genes were commonly regulated in the two extrathymic DP subsets, whereas 23 genes were uniquely expressed in MAT DPs, 31 different genes were exclusively expressed in aortic arch DPs. Conclusion: Our study is the first to identify and provide deep transcriptional analysis two extra-thymic DP populations in mice.

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Gene expression of CD4+CD8+ Tfh cells in lesions of IgG4-related disease and CD4+CD8+ Tfh in tonsil.

Project description:To further know features of CD4+CD8+ Double positive (DP)-Tfh cells in respective lymphoid tissues, we subsequently investigated transcriptomes of DP-Tfh cells in submandibular gland lesions of IgG4-RD in comparison to those of DP-Tfh cells in tonsils. Results showed that DP-Tfh cells of IgG4-RD lesions were more apt to present CTL-related genes such as Eomes and granzymes than DP-Tfh cells in tonsils. It was also noted that the level of CD70 in DP-Tfh cells of IgG4-RD lesions was higher than that of tonsillar DP-Tfh cells. Genes related to Tfh cell function were seemed to be presented in DP-Tfh cells of tonsils compared to DP-Tfh cells of IgG4-RD lesions.

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Bulk RNAseq analysis of CD4 CD8 double negative 3 (DN3) and CD4 CD8 double positive (DP) thymocytes in thymus autonomy and turnover

Project description:Bulk RNAseq analysis of CD4 CD8 double negative 3 (DN3) and CD4 CD8 double positive (DP) thymocytes in thymus autonomy and turnover

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miRNA expression in PTEN-deficient premalignant DP thymocytes

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