Project description:Goals/objectives: to identify various gene expression in B cell subsets derived from human PBMC and cord blood Sample: human PBMC and human cord blood Cell subsets: memory B cell (CD20+CD10-CD27+), naive B cell (CD20+CD10-CD27-), CD21lo transitional B cell (CD20+CD10+CD27-CD21lo), CD21hi transitional B cell (CD20+CD10+CD27-CD21hi) Number of sample: PBMC (pooled from 2 donors), cord blood (pooled from 2-3 donors) Number of repeats: 2 Microarray: Affymetrix HU133Plus2 RNA extraction: Trizol (Invitrogen) RNA preparation/hybridization: All according to affymetrix protocol
Project description:This SuperSeries is composed of the following subset Series: GSE10512: Gene Expression Profiles of PBMC Subsets in Acute GVHD Following Cord Blood Transplantation (IMS v.1) GSE10513: Gene Expression Profiles of PBMC Subsets in Acute GVHD Following Cord Blood Transplantation (IMS v.2) Keywords: SuperSeries Refer to individual Series
Project description:Transcriptome profiling was conducted on 2 replicate samples of total peripheral blood mononuclear cells (PBMC) and isolated PBMC subsets.
Project description:Microarray deconvolution is a technique for quantifying the relative abundance of constituent cells in a mixture based on that mixture's microarray signature and the signatures of the purified constituents. It has been applied to yeast and other systems but not to blood samples. Here we test the ability of this technique to determine the fractions of subsets of memory T cells in peripheral blood mononuclear cell (PBMC) samples. Experiment Overall Design: PBMC samples from several donors were split. One portion of each PBMC sample was set aside, while the remainder of each was sorted into naive, central memory, and effector memory T cell subsets.
Project description:High quality genetic material is an essential pre-requisite when analyzing gene expression using microarray technology. Peripheral blood mononuclear cells (PBMC) are frequently used for genomic analyses, but several factors can affect the integrity of nucleic acids prior to their extraction, including the methods of PBMC collection and isolation. In this study, we compared the Ficoll-Paque density gradient centrifugation and BD Vacutainer cell preparation tube (CPT) protocols to determine if either method offered a distinct advantage in preparation of PBMC-derived immune cell subsets for their use in gene expression analysis. We compared gene expression in PBMC and individual immune cell types from Ficoll and CPT isolation protocols using Affymetrix microarrays. PBMC from 3 healthy donors were isolated using Ficoll-Paque gradient or BD Vacutainer CPT methods. Subsequently, immune cell subsets were separated by positive selection from PBMC obtained by both methods. RNA was extracted from total PBMC isolated by both protocols and from their subsets, followed by analysis of gene expression using Affymetrix microarrays. Gene expression was compared between Ficoll and CPT samples for a given cell type.
Project description:High quality genetic material is an essential pre-requisite when analyzing gene expression using microarray technology. Peripheral blood mononuclear cells (PBMC) are frequently used for genomic analyses, but several factors can affect the integrity of nucleic acids prior to their extraction, including the methods of PBMC collection and isolation. In this study, we compared the Ficoll-Paque density gradient centrifugation and BD Vacutainer cell preparation tube (CPT) protocols to determine if either method offered a distinct advantage in preparation of PBMC-derived immune cell subsets for their use in gene expression analysis. We compared gene expression in PBMC and individual immune cell types from Ficoll and CPT isolation protocols using Affymetrix microarrays.
Project description:Human Naïve-like CD8 T cells induced by the Yellow Fever Vaccine 17D were compared to the conventional subsets in total CD8 T cells Samples originate from peripheral blood mononuclear cells (PBMC) from 8 different donors vaccinated with the YF-17D vaccine 1'000 cells from various CD8 T cells subsets were purified by flow cytometry, from 8 vaccinees (donors d1 to d8); the subsets (cell types) include: A2/NS4b tetramer positive CCR7+ CD45RA+ CD8 T cells (A2_NS4b Naïve-like), Total Naive (CCR7+ CD45RA+), Total Tscm (CCR7+ CD45RA+ CD58+ CD95+), Total CM (CCR7+ CD45RA-) and Total Effectors (CCR7 negative).
Project description:We exploited label-free quantitative mass spectrometry to compare primary human blood Dendritic cells (DCs) subsets protein expression to identify new markers. Subsets distinguished are: Plasmacytoid DCs (pDC) and BDCA3+ and CD1c+ myeloid DCs and CD16+ monocytes. The dendritic cells were analyzed by LC-MS/MS and processed by MaxQuant for identification and LFQ quantification.
Project description:Human PBMC, there are major T, B, Dc, Basophil, Eosinophil and Neutrophil subsets with typical biological functions. The characteristic behavior of each subset was determined by its unique gene expression and regulation. Difference between each subset or different treatment to the same subset will induce huge amount different gene expression. This will provide the basic evidence of understanding why and how each subset cells functions properly. We used microarrays to detail the global programme of gene expression in each subsets of human PBMC and identified distinctly up-regulated genes during this process.