Project description:Analysis of expression quantitative trait loci (eQTLs) using RNA derived from freshly harvested peripheral blood CD4+ lymphocytes from 200 asthmatics collected in clinical settings. RNA was obtained from peripheral blood CD4+ lymphocytes at four study centers. Peripheral blood (17 cc) was collected into BD Vacutainer CPT tubes (BD Diagnostics, Franklin Lakes, New Jersey) and placed on ice. Samples were centrifuged within 1 hour of collection for 20 minutes at 1700RCF, followed by mononuclear cell layer isolation and suspension in 10 ml of PBS. We isolated CD4+ lymphocytes using anti-CD4+ microbeads by column separation (Miltenyi Biotec, Auburn, CA) using 20 µl anti-CD4+ Micro beads per 106 total cells
Project description:Purpose:we'd like to provide the first DNA methylation profiling for ITP. Methods:Peripheral blood CD4+ T lymphocytes samples were collected from 4 primary refractory ITP cases and 4 age-matched healthy controls, and DNA methylome profiling was performed using Illumina Human Methylation850K. Results:The DNA methylome profiling identified a total of 260 differentially methylated CpG sites mapping to 72 hypermethylated and 64 hypomethylated genes. Conclusions:we performed genome-wide DNA methylation profiling of primary refractory ITP and healthy controls, and identified a set of differentially methylated genes and pathways.
Project description:Comparsion of the transcriptomes of naive, resting memory and activated memory CD4+ T lymphocytes. Peripheral blood lymphocytes were collected at baseline and day 13 post-inoculation of a healthy subject administered with vaccinia virus. Cells were isolated by FACs using antibodies against CD4, CD45RO and CD38.
Project description:Analysis of expression quantitative trait loci (eQTLs) using RNA derived from freshly harvested peripheral blood CD4+ lymphocytes from 200 asthmatics collected in clinical settings.
Project description:Gene expression data of primary human naive and memory CD4+T lymphocytes purified from peripheral blood are generated to be analyzed in different ways such as for traditional searching of differentially expressed genes between the two cell subsets or in combination to in-silico data of microRNAs target prediction for microRNAs known to be characteristically expressed in the two cell subsets.
Project description:The aim of this study was to characterize the transcriptional signature of MDR1+ human memory T cells isolated from clinically inflamed gut tissue, and compare it to local MDR1- memory T cells Human mononuclear cells were isolated from the peripheral blood of a healthy adult donor (Ficol density centrifugation) or from resected lesioned gut tissue of a patient with active Crohn's disease. For cell isolation from gut tissue, tissue was rinsed with PBS, treated with 0.15% DTT to remove mucous, then with 1 mM EDTA to remove epithelial cells and intra-epithelial lymphocytes. Remaining tissue was digested using Liberase-TL (Roche) plus 10 U/mL DNase I. Mononuclear cells were then filtered through 70 mM nylon filters and isolated via a 30%/ 70% Percol gradient. Mononuclear cells from blood or gut tissue were FACS-sorted into CD3+ CD4+ CD45RO+ MDR1+ or MDR1- memory T cells, using Rhodamine 123 (Rh123) efflux as a surrogate for MDR1 expression/ activity. Sorted cells were harvested directly ex vivo (without further in vitro culture or manipulation) prior to RNA extraction. MDR1- memory CD4+ T cells vs. MDR1+ memory CD4+ T cells from healthy donor peripheral blood or from active Crohn's disease lesioned tissue; MDR1- or MDR1+ memory CD4+ T cells from blood vs. inflamed gut tissue
Project description:The RNA-sequencing to profile patient peripheral blood mononuclear cell (PBMC) samples of systemic lupus erythematosus (SLE, N=25), primary Sjögren’s syndrome (pSS, N=23), and healthy control (HD, N=24). We collected on the steady-state and 18-hours anti-CD3 culture to identify the transcriptome remodeling upon T-cell stimulation. Each patient samples were sorted for CD4+ (Th cells) and CD8+ (CTL) T lymphocytes, CD16+CD7+ cells (NK cells), and CD19+ cells (B cells).
Project description:T lymphocytes are conventionally divided into subsets based upon expression of co-receptors, cytokines and surface molecules. By mRNA microarray analysis, T lymphocytes that express the C-type lectin CD161 were identified to share a transcriptional profile, which led to the identification of an innate function across these previously defined subsets, including CD8, CD4 and TCRgd T cells. Gene expression of magnetically enriched, resting CD161+ and CD161- CD4+ T cells from peripheral blood was measured in 3 healthy donors.
