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RNA-seq on human naive and memory regulaotry T cells before and after TCR stimulation.

ABSTRACT: Human FOXP3+ regulatory T cells were isolated as CD4+CD25+CD127- population from PBMCs. Cells were stimulated with TCR stimulation with the presence of rhIL-2. Naive cells were CD45RA+ and memory cells were CD45RA-.

INSTRUMENT(S): DNBSEQ-T7

ORGANISM(S): Homo sapiens

SUBMITTER: Yohei Sato

PROVIDER: E-MTAB-16435 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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RNA-seq on human regulaotry T cells before and after TCR stimulation

Project description:Human FOXP3+ regulatory T cells were isolated as CD4+CD25+CD127- population from PBMCs. Cells were stimulated with TCR stimulation with the presence of rhIL-2.

2025-06-01 | E-MTAB-14707 | biostudies-arrayexpress

Naive CD4+ T cell expansion defined by CD25 and CD31 expression

Project description:Naive CD4 T cells defined as (TCRab+ CD4+ CD45RA+ CD62L+ CD127+) were further divided by CD25 and CD31 expression with FACS-based flow sorting. Subsequently, gene expression levels of each subset were assessed by using transcription arrays.

2017-08-15 | E-MTAB-4853 | biostudies-arrayexpress

Microarray analysis of FoxP3-expressing or non-expressing subsets of human CD4+ T cells

Project description:Gene expression profiles of subsets of CD4+ T cells according to their expression of FoxP3 and CD45RA were compared. Abstract: FoxP3 is a key transcription factor for the development and function of natural CD4+ regulatory T cells (Tregs). Here we show that human FoxP3+CD4+ T cells are composed of three phenotypically and functionally distinct subpopulations: CD45RA+FoxP3low resting Tregs (rTregs) and CD45RA-FoxP3high activated Tregs (aTregs), both of which are suppressive in vitro, and cytokine-secreting CD45RA-FoxP3low non-suppressive T cells. The proportion of the three subpopulations characteristically altered in cord blood, aged individuals, and patients with immunological diseases. Terminally differentiated aTregs rapidly die while rTregs proliferate and convert into aTregs in vitro and in vivo as shown by the transfer of rTregs into NOD-scid-common gamma-chain-knockout mice and by TCR sequence-based T cell clonotype tracing in peripheral blood of normal individuals. Taken together, the dissection of FoxP3+ cells into subsets enables one to analyze Treg differentiation dynamics and interactions in normal and disease states, and to control immune responses through manipulating particular FoxP3+ subpopulations. RNA was extracted from freshly obtained peripheral blood lymphocytes from a healthy donor that were separated according to their expression of CD25, CD127 and CD45RA after surface staining.

2009-05-21 | E-GEOD-15659 | biostudies-arrayexpress

DNA Methylation Analysis of Systemic Lupus Erythematosus

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DNA Methylation Analysis of Systemic Lupus Erythematosus

Project description:This study performed Illumina Methylation450 analysis of CD4+ T-cells, CD19+ B-cells and CD14+ Monocytes from lupus patients and controls. A validation cohort was further analyzed with the same platform using CD4+ T-cells, CD45RO-CD45RA+ naive T-cells, CD45RO+CD45RA- memory T-cells, and CD25+CD127- regulatory T-cells. SLE v Control comparisions were made within each cell type. The SLE patient samples are labeled SLEnnnn and the controls Xnnnn. The cell type CD4, CD14, CD19, Tmem, Treg, Tnaive is appended to each sample ID.

2014-07-10 | E-GEOD-59250 | biostudies-arrayexpress

Transcription profiling of mouse naive CD4+CD25-CD62Lhi T cells and nTreg cells with Eos knock-down

Project description:Eos expression in Treg cells have been documented by several microarrays including ours. We hypothesized that Eos facilitates Foxp3- dependent gene repression in Regulatory T cells. In order to investigate the role of Eos in mediating the Foxp3-dependent gene silencing program, we utilized lentiviral shRNA knockdown of Eos in natural Tregs isolated from the periphery of Balb/C mice. A renilla luciferase (RL) gene specific shRNA lentivirus was used as a control for the transduction of cells. The transcriptional profile of naive T cells, natural Tregs, Eos knockdown Tregs, and control shRNA knockdown Tregs was compared using Agilent 4x 44K whole mouse genome array. The goal of this microarray is to document the global effect of the loss of Eos expression on the transcriptional profile of natrual Treg cells. Experiment Overall Design: Eos knock-down (si-Eos) was mediated by Lentivirus expressing GFP as a reporter of transduction and sorting marker. Renilla luciferase specific shRNA lentiviral transduction was carried out in parallel as a control. NaÃ¯ve (CD4+CD25-CD62Lhi) T cells and nTreg cells were freshly isolated from Balb/C mice.

2009-10-16 | E-GEOD-17166 | biostudies-arrayexpress

CD4+ cell transcriptional profiling by RNA sequencing

Project description:<p>We use next generation sequencing to investigate the different transcriptomes of closely related CD4+ T-cells from healthy human donors to elucidate the genetic programs that underlie their specialized immune functions. Six cell types were included: Regulatory T-cells (CD25hiCD127low/neg with >95% FOXP3+ purity), regulatory T-cells activated using PMA/ionomycin, CD25-CD45RA+ ('naive' helper T-cells), CD25-CD45RO+ ('memory' helper T-cells), activated Th17 cells (>98% IL17A+ purity) and activated IL17-CD4+ T-cells (called 'ThPI'). Poly-T capture beads were used to isolate mRNA from total RNA, and fragment sizes of ~200 were sequenced from both ends on Illumina's genome analyzer. We confirm many of the canonical signature genes of T-cell populations, but also discover new genes whose expression is limited to specific CD4 T-cell lineages, including long non-coding RNAs. Additionally, we find that genes encoded at loci linked to multiple human autoimmune diseases are enriched for preferential expression upon T-cell activation, suggesting that an aberrant response to T-cell activation is fundamental to pathogenesis.</p>

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Profiling of naÃ¯ve and memory Tregs

Project description:Changes in Treg function are difficult to quantify due to the lack of Treg-exclusive markers in humans and the complexity of functional experiments. We sorted naive and memory human Tregs and conventional T cells, and identified genes that identify human Tregs regardless of their state of activation. We developed this Treg signature using Affymetrix human genome U133A 2.0 microarrays. To generate Tregs and Tconvs in multiple states of activation, naÃ¯ve (CD4+CD25hiCD45RA+) and memory (CD4+CD25hiCD45RA-) Tregs, and naÃ¯ve (CD4+CD25-CD45RA+) and memory (CD4+CD25-CD45RA-) Tconvs were sorted from blood of 7 healthy adults and RNA was isolated ex vivo or after stimulation for 40h, promoting activation-induced FOXP3 in Tconvs. The gene-expression profile of the eight cell subsets was analyzed. 7 adult healthy control samples were sorted into 4 subsets: naÃ¯ve (CD4+CD25hiCD45RA+) and memory (CD4+CD25hiCD45RA-) Tregs, and naÃ¯ve (CD4+CD25-CD45RA+) and memory (CD4+CD25-CD45RA-) Tconvs. These were used for RNA ex vivo and after 40h stimulation with anti-CD3/CD28 beads to induce an activation phenotype.

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Single cell transcriptomic analysis of human CD25+ CD127- CD4+ Treg cells and CD25- CD127+ CD4+ Tconv cells isolated from peripheral blood

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2018-03-15 | GSE111901 | GEO

Expression data from human effector CD4 T cells co-cultured with various Treg subsets sorted from human peripheral blood.

Project description:Teff cells (CD4+CD25-CD127+CD45RO+) were co-cultured with and without sorted Treg subsets plus dendritic cells for 18h. All the sorted Teff cells are gated on CD4+CD25-CD127(hi), and further sorted for CD45RO- and CD45RO+ phenotype.

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