Healthy volunteers gene expression study of CD4 and NK lymphocytes
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ABSTRACT: Sixteen volunteers, that were age, gender and BMI matched, were included in the study. Blood samples were taken one per each volunteer on three different days to also assess variability of gene expression markers in one individual. In all datasets variability between volunteers was higher than variability of measurables in samples collected from the same volunteer on different days. We have identified genes in individuals that are highly stable during one month period and are thus suitable as markers of individual disease/therapy progress. A list of genes with low overall variability is also available. Interestingly, data show that important components of immune response are down-regulated with BMI in T helper and NK cells. This paper also gives the first insight into the gene expression profile of T helper and NK cells in healthy population. CD4 and NK lymphocytes were isolated with positive selection using magnetic nanoparticle based kits from StemCell Inc. (Canada) and frozen to -80°C until total RNA extraction.
Project description:Sixteen volunteers, that were age, gender and BMI matched, were included in the study. Blood samples were taken one per each volunteer on three different days to also assess variability of gene expression markers in one individual. In all datasets variability between volunteers was higher than variability of measurables in samples collected from the same volunteer on different days. We have identified genes in individuals that are highly stable during one month period and are thus suitable as markers of individual disease/therapy progress. A list of genes with low overall variability is also available. Interestingly, data show that important components of immune response are down-regulated with BMI in T helper and NK cells. This paper also gives the first insight into the gene expression profile of T helper and NK cells in healthy population.
Project description:In this experiment we generated Affymetrix gene expression data for T Follicular Helper (TFH) cells from tonsils of healthy volunteers (4 biological replicates) and naive CD4-positive helper T cells (2 biological replicates). TFH cells provide a model relevant to SLE as TFH operate upstream of the activation of pathogenic autoantibody-producing B cells during the disease. This experiment accompanies promoter capture-C and ATAC-seq experiments on the same cell types.
Project description:To study the effect of paracrine interleukin-9 (IL-9) on T helper cells, we incubated IL-9R+ T helper cells isolated from blood (5 samples) and skin (3 samples) with or without IL-9 for 12h and the transcriptome was analyzed by bulk RNA-seq.
Project description:The diversity of human immune responses to P. falciparum is unknown and yet immune decision-making likely dictates outcome of infection We infected 15 malaria-naïve human volunteers with P. falciparum and used longitudinal whole blood transcriptional profiling to independently analyse the immune response in every volunteer
Project description:NK cells may acquire under certain conditions features of adaptive immune cells. As the functional role of memory NK cells in cancer has so far remained elusive, we reasoned whether tumor-priming itself might promote memory NK cell generation. We provide substantial evidence that independent from pro-inflammatory stimulation, tumor-induced memory-like (TIML) NK cells exhibit a heightened, tumor-restricted cytotoxicity which is dependent on a higher/faster perforin but not IFN-γ release. Comparative transcriptome analysis reveals that gene expression patterns differ between TIML- and Cytokine-induced memory-like (CIML)-NK cells.
Project description:Comparison of gene expression from subjects who resolved or formed pustules to H.ducreyi. In human inoculation experiments, the cutaneous immune response to Haemophilus ducreyi consists of serum, PMN, macrophages, T cells and myeloid DC. In reinfection experiments, some subjects form pustules twice (PP group) or resolve infection twice (RR group). Although pustule formation is associated with serum resistance and phagocytic failure, there are no differences in the ability of isolated phagocytes or serum obtained from PP and RR subjects to ingest or kill H. ducreyi. To identify the basis for differential host susceptibility to H. ducreyi, we used microarrays to profile gene expression in infected and uninfected tissue and monocyte-derived DC obtained from PP and RR subjects. In infected tissue, both groups had a core response to H. ducreyi. Many additional transcripts that signify active immune function were upregulated exclusively in RR tissue, while PP tissue exclusively contained differentially regulated transcripts consistent with immune dysregulation. The core response of infected DC from both groups was typical of a DC1 response. RR DC exclusively expressed many additional transcripts indicative of DC1 function, while PP DC uniquely expressed differentially regulated transcripts characteristic of both DC1 and DCreg. The data suggest that DC from PP and RR subjects are prewired to respond differentially to H. ducreyi. Experiment Overall Design: Six healthy adult volunteers (5 females, 1 male, 36 ± 13 yrs, mean age ± SD) who had been infected with H. ducreyi twice were inoculated a third time. Each volunteer was inoculated at 3 sites on the upper arm with live H. ducreyi 35000HP (a human passaged isolate of strain 35000) and at 1 site with sterile PBS. Forty-eight hours after inoculation, lesion size was measured and RNA was isolated from the infected site that had the largest diameter. RNA was also obtained from the PBS control site (uninfected site). We obtained peripheral blood 6 to 12 months after inoculation of the 6th subject and derived myeloid DC in vitro. In brief, CD14+ peripheral blood monocytes were isolated from PBMC by positive selection magnetic beads (Miltenyi Biotec, Auburn, CA) and grown in the presence of recombinant human (rh) IL-4 (1 ng/ml) and rhGM-CSF (0.2 ng/ml) (R&D Systems, Minneapolis, MN). The cells were HLA-DR+, CD86+ CD40+, CD3-, CD14-, and CD19- by flow cytometry. The experiment was done in pairs so that DC from 1 RR subject were exposed to the same inoculum as DC from 1 PP subject. DC were incubated with nonopsonized H. ducreyi for 90 minutes at an MOI of 30:1. The DC were washed to remove non-associated bacteria, and incubated an additional 22.5 hours. Cells were collected and used for microarray analysis. Supernatants were collected and analyzed for cytokines using the Human Th1/Th2 II Cytometric Bead Array Kit per manufacturerâs instructions (BD Biosciences).
Project description:Background: Although genome-wide association studies (GWAS) have identified hundreds of variants associated with risk of autoimmune and immune-related disorders (AID), our understanding of the diseases mechanisms is limited. In particular, more than 90% of the risk variants lie in non-coding regions, and almost 10% of these map to long non-coding RNA transcripts (lncRNAs). LncRNAs are known to show more cell-type specificity than protein-coding genes. Methods: In this study, we aimed to characterize lncRNAs and protein-coding genes located in loci associated with nine AID which have been well-defined by Immunochip analysis, by transcriptome analysis across seven peripheral blood leukocyte populations (granulocytes, monocytes, NK cells, B-cells, memory-T cells, naive CD4+ and naive CD8+ T-cells) and four cord blood derived T-helper cell populations (precursor, primary, polarized (Th1, Th2) T-helper cells). Results: We show that lncRNAs mapping to loci shared between AIDs are significantly enriched in immune cell types when compared to lncRNAs from the whole genome (M-NM-1<0.005). We were not able to prioritize single cell types relevant for specific diseases, but we observed five cell types enriched (M-NM-1<0.005) in five AID (NK cells for inflammatory bowel disease, juvenile idiopathic arthritis, primary biliary cirrhosis, psoriasis; memory-T and CD8+ T-cells in juvenile idiopathic arthritis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis; Th0 and Th2 cells for inflammatory bowel disease, juvenile idiopathic arthritis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis). Furthermore we show that co-expression analyses of lncRNAs and protein-coding genes can predict the signaling pathways in which these AID-associated lncRNAs are involved. Conclusions: The observed enrichment of lncRNA transcripts in AID loci implies an important role for lncRNAs in AID etiology and suggests that lncRNA genes should be studied in more detail to correctly interpret GWAS findings. The co-expression results strongly support a model in which the lncRNA and protein-coding genes function together in the same pathways. 7 immune cell types
Project description:Cutaneous sarcoidosis skin provides relatively non invasive access to granulomatous sarcoidosis tissue. Twenty participants were enrolled: 15 with active CS and 5 healthy volunteers. Microarray analyses comparing non-LS and healthy volunteer skin with LS showed several thousand genes differentially expressed
Project description:Baseline gene expression in circulating peripheral blood mononuclear cells isolated from 18 apparently healthy volunteers (10 females and 8 males). The volunteers fasted overnight for 10 hours prior to collection of blood samples (but they were allowed water). 5 blood samples were collected from each volunteer at consecutive 8-day intervals. Volunteers were included in this study provided they met the following selection criteria, that they should be <br>?Aged between 20 and 45 years of age; <br>?A non-smoker (or an ex-smoker);<br>?Not pregnant or been pregnant in the last 12 months <br>?Not diagnosed with any long-term medical condition; <br>?Or have not (in the previous 4 months) donated blood, received any inoculations and are not taking regular prescribed medicine (including HRT and oral contraception) <br>Prior to inclusion in this study volunteers were screened for general health and had a blood sample taken for routine screening of Hb content etc. Only those volunteers who were in good general health were allowed to participate in the study. <br>The age of all volunteers has been reported here as being 20-45 years in order to preserve the anonymity of the data, and so that the individuals involved may not be able to track down their own specific data. Please contact us if the details of volunteers' ages are required.
Project description:We report the enhancer landscape in primary human macrophages and foam cells using ChIP-seq for the H3K27ac histone mark CD14+ monocytes were isolated from the blood of 2 healthy male volunteers. Monocytes were differentiated into macrophages by culture for 7 days with 50ng/ml macrophage colony stimulating factor and then treated for 48 hours with either oxidized low density lipoprotein (oxLDL) to induce foam cell formation or with a control buffer that lacked oxLDL. The resulting 4 samples were then subjected to ChIP-seq for H3K27ac.