Project description:We compared differences in fetal and adult T cells by performing whole genome profiling on sort-purified T cells (naïve CD4+ and Treg cells) from human fetal specimens (18-22 gestational weeks) and adult specimens (age 25-40 years old). Fetal and Adult Naïve CD4+ T cells phenotype: CD3+CD4+CD45RA+CCR7+CD27+, Fetal and Adult CD4+CD25+ Treg phenotype: CD3+CD4+CD25bright Four different groups were analyzed: Fetal Naïve CD4+ T cells, Adult Naïve CD4+ T cells, Fetal Treg cells, Adult Treg cells. For each group three independent donors were analyzed.
Project description:Cells were isolated from healthy human donors (n=2). Unstimulated cells. Cells were stained with CD4, CD45RA, CCR7 and CXCR7. Using flow cytometry, 4 CD4+ T cell populations were sorted: (1) Naïve (CD45RA+CCR7+CXCR5-), (2) Central memory (CD45RA-CCR7+CXCR5-), (3) Effector memory (CD45RA-CCR7-CXCR5-) and (4) CXCR5+ cells (CD45RA-CCR7-CXCR5+) RNA were extracted from sorted cells and hybridized on Affymetix HU133Plus2 chips. Each population were repeated on different donors such that n=2.
Project description:Cells were isolated from healthy human donors (n=2). Unstimulated cells. Cells were stained with CD4, CD45RA, CCR7 and CXCR7. Using flow cytometry, 4 CD4+ T cell populations were sorted: (1) Naïve (CD45RA+CCR7+CXCR5-), (2) Central memory (CD45RA-CCR7+CXCR5-), (3) Effector memory (CD45RA-CCR7-CXCR5-) and (4) CXCR5+ cells (CD45RA-CCR7-CXCR5+)
Project description:Buffy coats from four independent human donors were enriched for CD4+ cells using MACS CD4 beads and LS columns. Cells were subsequently sorted into Naïve, Central Memory and Effector Memory using following markers: Naive: CD4+CD25–CD45RA+CCR7+; Central Memory: CD4+CD25–CD45RA–CCR7+; Effector Memory: CD4+CD25-CD45RA-CCR7-. Total RNA from these cell subsets was extracted and 100ng was used for Nanostring SPRINT run according to manufacturer's instructions. Overall aim of the experiment is assessing the expression level of human T helper cell miRNome.
Project description:We compared differences in fetal and adult T cells by performing whole genome profiling on sort-purified T cells (naïve CD4+ and Treg cells) from human fetal specimens (18-22 gestational weeks) and adult specimens (age 25-40 years old). Fetal and Adult Naïve CD4+ T cells phenotype: CD3+CD4+CD45RA+CCR7+CD27+, Fetal and Adult CD4+CD25+ Treg phenotype: CD3+CD4+CD25bright
Project description:Ascertain the effects of disease-causing gene mutations on the differentiation status of human naïve CD4+ T cells in the setting of primary immunodeficiencies. Thus, do CD4+ T cells isolated according to a naïve surface phenotype (ie CD4+CD45RA+CCR7+) from healthy donors exhibit a similar gene expression profile as phenotpyically-matched cells isolated from individuals with defined primary immunodeficiencies caused by specific monogenic mutations.
Project description:T-cell replete cord blood transplantation results in a rapid thymus-independent T-cell reconstitution which is strikingly CD4+ biased compared to the well-established observation of CD8+ T-cell biased expansion after T-cell replete bone marrow transplant. We used microarrays to detail the distinct gene expression profile underlying rapid CD4+ T cell expansion following T-cell replete cord blood transplantation and identified distinct up-regulated pathways during this process.
Project description:Human CD8+ T cells are functionally heterogeneous and can be divided into distinct subsets according to CCR7 and CD45RA expression levels. Among the subsets, CCR7-CD45RA+ CD8+ T cells are considered to be terminally differentiated cells and designated as Temra. Temra show attenuated ability to proliferate and produce IFN-gamma in response to TCR stimulation, while Temra show improved function after IL-15 treatment. To clarify the transcriptional signatures induced by the stimulations, Temra were purified using flow cytometry, stimulated with IL-15 or with anti-CD3/CD28 (TCR stimulation), or cultured without stimulation for 2 days, and subjected to microarray analysis.
Project description:Infection with cytomegalovirus is characterised by very strong and sustained T cell responses in peripheral blood. These expansions increase even further with age and research suggests CMV-specific T cells may contribute towards development of accelerated immune senescence and vascular disease in elderly people. In this study we compared the transcriptional profile of CD4+ T cells specific for two CMV-derived epitopes to that of CD4+CCR7-CD45RA- (effector memory) T cells of CMV-seronegative individuals. The aim was to get a better understanding of the nature of these virus-specific T cells and how they may contribute to immunopathology.
Project description:To examine the broad impact of IL-27 on human T lymphocytes, we performed a microarray analysis assessing >20,000 well annotated genes on purified naïve (CD45RA+CD45RO-CCR7+) and central memory (CD45RA-CD45RO+CCR7+) CD4+ and CD8+ T cells from three healthy donors that were activated in vitro (plate bound anti-CD3 and soluble anti-CD28) in the presence or absence of human recombinant IL-27 (100 ng/mL). Our goal was to investigate the impact of interleukin-27 on the gene expression profil of human CD4 and CD8 T lymphocytes.