Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Expression data from cord blood derived CD4+ CD25+ (Treg) and CD4+ CD25- (Thelper) cells, both while resting & after stimulation

ABSTRACT: Here we compare the effects of stimulation on cord blood derived CD4+ CD25+ (Treg) and CD4+ CD25- (Thelper) cells, isolated by MACS protocols & expanded in vitro using dynabeads. Expansion was carried out at a ratio of 3 beads/cell in the presence of 1000units/ml of recombinant human IL2 for 8 days, followed by 3 days of culture without beads. RNA was extracted from resting cells on day 4 after expansion. Stimulated cells were treated with Ionomycin for 2hrs prior to RNA extraction. Four way comparison of resting and stimulated Tregs versus Thelpers. A total of 18 arrays were analysed containing RNA from 5 individual cord blood donors.

ORGANISM(S): Homo sapiens

SUBMITTER: Steve Pederson

PROVIDER: E-GEOD-20934 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Genome-wide identification of human FOXP3 target genes in natural regulatory T cells.

Sadlon Timothy J TJ Wilkinson Bridget G BG Pederson Stephen S Brown Cheryl Y CY Bresatz Suzanne S Gargett Tessa T Melville Elizabeth L EL Peng Kaimen K D'Andrea Richard J RJ Glonek Gary G GG Goodall Gregory J GJ Zola Heddy H Shannon M Frances MF Barry Simon C SC

Journal of immunology (Baltimore, Md. : 1950) 20100616 2

The transcription factor FOXP3 is essential for the formation and function of regulatory T cells (Tregs), and Tregs are essential for maintaining immune homeostasis and tolerance. This is demonstrated by a lethal autoimmune defect in mice lacking Foxp3 and in immunodysregulation polyendocrinopathy enteropathy X-linked syndrome patients. However, little is known about the molecular basis of human FOXP3 function or the relationship between direct and indirect targets of FOXP3 in human Tregs. To in ...[more]

PMID: 20554955

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Project description:We have previously developed an approach that fractionates genomic DNA fragments depending on their CpG density (methyl-CpG-immunoprecipitation, MCIp), and adapted this approach to identify regions that are differentially methylated in the two closely related regulatory T-cells (Treg cells) and conventional T-cells (Tconv cells). Because Treg cells naturally occur at a relatively low frequency, we used a previously established protocol to expand Treg cells from a stable naÃ¯ve Treg population that is characterized by the co-expression of CD4, CD25 and CD45RA. We separated gDNA of both expanded T cell lineages (Tregexp and Tconvexp) into unmethylated (CpG) and methylated pools (mCpG) using MCIp and compared cell type-specific differences in DNA methylation by co-hybridization of the two umethylated or the two methylated DNA subpopulations of Treg and Tconv, respectively, to these locus-wide custom tiling arrays. As enriched DNA-fragments from a cell type in the methylated fraction should be depleted in the unmethylated fraction, the signal intensities in CpG pool and mCpG pool hybridizations should complement themselves (âMirror-Imageâ approach) and thereby allow the identification of differentially methylated regions (DMR). Because we expected to find lineage-specific methylation differences with greater probability in regions associated with differential transcriptional activity, we limited our analysis to gene loci that showed cell type-specific gene expression in Treg versus Tconv cells plus a handful of control regions that were equally expressed in both cell types. The microarray used in this study covered 12 megabases of the human genome and contained 69 regions (with a median size of 100.000 kb) and 128 proximal promoter regions and 181 genes, which included a number of well known and functionally relevant genes like CD40LG, IFNG, FOXP3, IL2RA and CTLA4. Keywords: MCIp-on-chip; comparative genomic hybridization With MCIp gDNA from Treg or Tconv cells was separated into hypo- and hypermethylated pools. On each array, wheather the two hypomethylated fractions- one from Treg, the other from Tconv cells- or the two hypermethylated fractions were cohybridized. Two biological replicates.

Dataset Information

Expression data from cord blood derived CD4+ CD25+ (Treg) and CD4+ CD25- (Thelper) cells, both while resting & after stimulation

Publications

Genome-wide identification of human FOXP3 target genes in natural regulatory T cells.

Similar Datasets

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