MicroRNA expression profiling of PPD stimulated bovine PBMC from M. bovis-challenged control and BCG vaccinated cattle
Ontology highlight
ABSTRACT: Micro RNA profiling was performed on in vitro PPDB and nil stimulated bovine PBMCs isolated from BCG vaccinated and unvaccinated cattle before and after challenge with M. bovis.
Project description:The process of hepatocarcinogenesis in the diethylnitrosamine (DEN) initiation/phenobarbital (PB) promotion mouse model involves the selective clonal outgrowth of cells harboring oncogene mutations in Ha-ras, B-raf, or Ctnnb1. Here, we have characterized mouse liver tumors harboring either Ctnnb1 or Ha-ras mutations via integrated molecular profiling at the transcriptional and translational and post-translational levels. In addition, metabolites of the intermediary metabolism were quantified by high resultion 1H magic angle nuclear magnetic resonance (HR-MAS NMR). We have identified tumor characteristic genotype-specific differences in mRNA and miRNA expression, protein levels, and post-translational modifications and in metabolite levels that facilitate the molecular and biochemical stratification of tumor phenotypes. Bioinformatic integration of these data at the pathway level led to novel insights into tumor genotype-specific aberrant cell signaling and in particular to a better understanding of alterations in pathways of the cell intermediary metabolism, which are driven by the constitutive activation of the β-Catenin and Ha-ras oncoproteins in tumors of the two genotypes. Male C3H/HeJ mice received a single i.p. injection of DEN (10 or 90µg/g body weight) at 2, or 6 weeks of age. After a treatment-free interval of 2 weeks, the C3H/HeJ mice were either kept on a diet containing 0.05% PB or on a PB-free control diet for 28 to 36 weeks before they were sacrificed. Ha-ras- or Ctnnb1-mutated tumors and control tissues were isolated and either flash frozen in liquid nitrogen and stored at -80°C, or prepared for immunohistochemistry.
Project description:Single cell ATAC-seq of PBMC - resting and stimulated. Used for comparison to asses the capabilies of the five-prime sequencing method in the detection of cis-regulatory elements using SCAFE (see publication).
Project description:IRF5 and STAT4 are strongly associated with human systemic lupus erythematosus (SLE). By performing chromatin immunoprecipitation-sequencing (ChIP-Seq) in human peripheral blood mononuclear cells (PBMCs), we identified more than 7000 target genes for IRF5 and STAT4 in stimulated PBMCs. Contrarily, without stimulation IRF5 seemed to be inactive, and STAT4 only showed low levels of transcriptional regulatory activity. Target genes of IRF5 and STAT4 were identified in human PBMCs with or without stimulation using ChIP-Seq.
Project description:Gene expression profiles in PPD-stimulated and unstimulated peripherhal blood mononuclear cells isolated from chinese cynomolgus macaques before and after BCG vaccination and before and after M. tuberculosis challenge were compared in animals able to control TB infection and those that developed TB disease in order to identify potential correlates of protection and/or biomarkers of disease. 6 macaques received BCG vaccination prior to challenge and were able to control TB infection (vaccinated controllers). 3 unvaccinated macaques were also able to control TB infection after challenge (unvaccinated controllers) and 3 other unvaccinated macaques developed TB disease and reached humane endpoint criteria (unvaccinated progressors). PBMCs isolated at 8 and 18 weeks post BCG-vaccination from the vaccinated controllers and at 6 weeks post M.tb challenge and at 2 time-points when the animals were naive in all animals were stimulated with PPD. RNA was extracted from the cells and hybridised to and Agilent rhesus macaque GE microarray in a one-colour hybridisation. Gene expression post-vaccination and/or post-challenge was compared with expression before vaccination/challenge when the animals were naive.
Project description:Tiling arrays have been reported as highly relevant tools to improve annotation and identify new transcripts in genome sequences. Our aim was to refine annotation of the Swine Leucocyte Antigen (SLA) complex, which is a gene dense region spanning 2.4 megabases on both sides of the centromere of chromosome 7 with many genes involved in immune response and inflammation. To date, one hundred and fifty-one SLA genes and pseudogenes have been characterized or predicted. A high-density tiling array was produced, which covered the entire Hp1a.0 haplotype sequence and partially the centromeric junction on the q-arm. The array contained 386,620 55-70-mer oligonucleotides designed on both strands and tiled on average every nine bases. It was used to compare the transcriptome of peripheral blood mononuclear cells (PBMCs) either mock-stimulated or stimulated with phorbol myristate acetate (PMA) and ionomycin for 24 hours. In both conditions, 19% of the probes were expressed among which 47% not previously annotated. In addition to the expected transcripts, segmentation analysis revealed new transcriptional architectures of previously annotated genes, a limited transcription activity in the centromeric junction and frequent transcription in antisense orientation. Differential expression of non-annotated transcripts was detected between non-stimulated and stimulated PBMCs, suggesting a regulatory role during immune cell activation. These results contribute to drawing an extensive transcription map of the SLA complex and to comparative functional mapping between mammals. This kind of approach can be applied to any candidate region of the pig genome in order to carry out a consistent functional annotation. Two-condition experiment, PMA ionomycine stimulated PBMCs vs. Control PBMCs. Biological replicates: 1 control, and 1 PMA ionomycine stimulated from 4 animals independently grown and harvested. One replicate per array. Total 8 slides.
Project description:Gene expression microarray data in resting myeloid cells and CD4+ T cells, as well as in response to lipopolysaccharide (LPS) or phytohemagglutinin (PHA) stimulation for 24 hours, respectively. Myeloid cells and CD4+ T cells were purified from resting and stimulated peripheral blood mononuclear cells from cord blood samples of 152 individuals at birth. Raw array data and quantile normalised gene expression data are available. Full summary statistics of eQTL analysis and interaction tests for response eQTLs are available. Information such as sample attributes and experimental variables are available in the \\"Microarray_sample_info.txt\\".
Project description:Background: This study focuses on the analysis of miRNAs expression data in a cohort of 181 well characterised breast cancer samples composed primarily of triple-negative (ER/PR/HER2-negative) tumours with associated genome-wide DNA and mRNA data, extensive patient follow-up and pathological information. Results: We identified 7 miRNAs with a prognostic role in the triple-negative tumours and an additional 8 prognostic miRNAs when the analysis was extended to the set of all ER-negative cases. miRNAs linked to an unfavourable prognosis were associated with a broad spectrum of motility mechanisms involved in the invasion of stromal tissues, such as cell-adhesion, growth factor-mediated signalling pathways, interaction with the extracellular matrix and cytoskeleton remodelling. When we compared different intrinsic molecular subtypes we found 46 miRNAs that were specifically expressed in one or more intrinsic subtypes. Integrated genomic analyses indicated these miRNAs to be largely influenced by DNA genomic aberrations and to exert a silencing effect on their targets through transcriptional down-regulation. Among others, our analyses highlighted the role of miR17-92 and miR-106b-25, two polycistronic miRNA clusters with known oncogenic functions. We showed that their basal-like specific up-regulation is influenced by increased DNA copy number and contributes to the transcriptional phenotype and the activation of oncogenic pathways in basal-like tumours.Conclusions: This is the first study analysing miRNA, mRNA and DNA data in integration with pathological and clinical information, in a large and well-annotated cohort of triple-negative breast cancers. It provides a conceptual framework, as well as integrative methods and system-level results to elucidate the role of miRNAs as biomarkers and modulators of oncogenic processes in these types of tumours. 181 breast tumour samples were analyzed, extracted from 173 patients. For the great majority of patients (165) only one sample was extracted, while for 8 patients two samples were extracted (biological replicates).
Project description:To identify microRNA changes during plasmacytoid dendritic cell (PDC) activation, we stimulated human primary PDCs with 10ug/ml R837 (Invivogen, San Diego, CA, USA) for 4 hours. Purified human pDCs were divided into two parts: one was cultured with medium alone, another was cultured with R837. 4 hours later, cells were collected and total RNA was extracted for the TaqManM-BM-. Human MicroRNA Arrays. The experiment was duplicated (sample1 and sample2).
Project description:Allergen-stimulated T cells from henâs egg-allergic children were analyzed to identify genes that are specifically up-regulated in these cells. Experiment Overall Design: PBMCs from three henâs egg allergic children and two non-allergic children were cultured in RPMI 1640 supplemented with 10% autologous plasma for 16 hours with or without henâs egg white allergen (allergen concentration10 mg/ml). To focus on specific T cells reacting with HEW, cells bearing CD4 antigen were isolated from cultured PBMCs using a Magnetic cell sorterï¼Miltenyi Biotec, Bergisch Gladbach, Germanyï¼after CD14 positive cell depletion. Total RNA was extracted from these CD4 positive cells with an RNeasy mini-kit (Qiagen, Valencia, CA, USA) according to the manufacturerâs instruction.