Project description:Background: There is ample evidence of blood-born miRNA signatures for various human diseases. To dissect the origin of disease-specific miRNA expression in human blood cells, we separately analyzed the miRNome of eosinophilic and neutrophilic granulocytes, monocytes, B-cells, T-cells, and natural killer cells, each in lung cancer patients and healthy individuals. Results: We found specific miRNA expression patterns for each immune cell type and also depending on the cell origin, line of defense, and function. The overall expression pattern of each leukocyte subtype showed great similarities between lung cancer patients and healthy controls. However, for each cell subtype we identified miRNAs that were deregulated in lung cancer patients including hsa-miR-21, a well- known oncomiR associated with poor lung cancer prognosis that was up-regulated in all subtype comparisons of lung cancer versus controls. While the miRNome of cells of the adaptive immune system allowed only a weak separation between patients and controls, cells of the innate immune system allowed perfect or nearly perfect classification. Conclusions: Leukocytes of lung cancer patients show a cancer-specific miRNA expression profile. Our data also show that cancer specific miRNA expression pattern of whole blood samples are not determined by a single cell type. The data indicate that additional blood components, like erythrocytes, platelets, or exosomes might contribute to the disease specificity of a miRNA signature.
Project description:Diversity of biological molecules in newborn and adult immune cells contributes to differences in cell function and atopic properties. Micro RNAs (miRNAs) are reported involve in the regulation of immune system. Therefore, determining the miRNA expression profile of leukocyte sub-populations is important for understanding immune system regulation. In order to explore the unique microRNA profiling that contribute to altered immune in neonates, we comprehensively analyzed the functional miRNA signatures of eight leukocyte subsets (polymorphonuclear cells, monocytes, CD4+ T cells, CD8+ T cells, natural killer cells, B cells, plasmacytoid dendritic cells (pDCs), and myeloid dendritic cells (mDCs)) from both neonatal and adult umbilical cord and peripheral blood samples, respectively. We observed distinct miRNA profiles between adult and neonatal blood leukocyte subsets, including unique miRNA signatures for each cell lineage. Leukocyte miRNA signatures were altered after stimulation. Adult peripheral leukocytes had higher let-7b-5p expression levels compared to neonatal cord leukocytes across multiple subsets, irrespective of stimulation. Transfecting neonatal monocytes with a let-7b-5p mimic resulted in a reduction of LPS-induced IL-6 and TNF-a production, while transfection of a let-7b-5p inhibitor into adult monocytes enhanced IL-6 and TNF-a production. With this functional approach, we provide intact differential microRNA expression profiling of specific immune cell subsets between neonates and adults. These studies serve as a basis to further understand the altered immune response observed in neonates and advance the development of therapeutic strategies
Project description:Gene expression microarray profiling was performed on peripheral blood leukocyte subsets (CD4+ T cells, CD14+ monocytes, and CD16+ neutrophils) from healthy controls and patients with flaring autoimmune disease.
Project description:We performed single-cell RNA sequencing (scRNASeq) on cryopreserved peripheral blood mononuclear cells (PBMCs) of healthy controls and OTULIN haploinsufficient patients in order to clarify the perturbation, if any, of cellular phenotypes in various leukocyte subsets in vivo at basal state attributable to OTULIN haploinsufficiency.
Project description:Circulating miRNAs has recently emerged as clinically relevant blood-based biomarkers for disease detection, tracking and prediction. The stability of these species combined with easy accessibility in circulation makes them attractive candidates for rapid and economic surveillance of broad spectrum disorders requiring invasive diagnosis.In this work we directly assess the utility of non-invasive blood-based biomarkers as an alternative strategy to accurately predict incidences of Ulcertaive Colitis. Whole genome miRNA expression levels in Microvescicles , Peripheral Blood Mononuclear Cells (PMBC) and platelets from a cohort of 20 Ulcerative Colitis patients and 20 normal individuals were measured using Affymetric microarrays. 20 cases/controls in three fractions
Project description:This project aims at the detection of specific patterns of miRNAs in peripheral blood samples of lung cancer patients. As controls, blood of donors without known affection have been tested. Using the miRNA patterns we hope to detect a diagnostic pattern for the non-invasive diagnosis of non-small cell lung carcinoma.
Project description:Affymetrix miRNA arrays were used to generate miRNA profiles of peripheral blood leukocytes and FACS sorted neutrophils, monocytes, B-cells, T-cells, CD4+ T-cells, and CD8+ T-cells, being the major leukocyte cell types in human. The study allowed for the determination of the miRNAs that were expressed in each leukocyte cell subtype. Two-way hierarchical clustering on the miRNAs and samples illustrated that miRNA expression profiles of B- and T-cells were very much alike, and that there there were a number of miRNAs which appeared to have an expression profile specific to certain leukocyte cell subtypes. This study will facilitate the identification of microRNAs associated with and contributing to single leukocyte cell subtypes. Peripheral blood leukocytes were extracted, and neutrophils, monocytes, B-cells, T-cells, CD4+ T-cells, and CD8+ T-cells were FACS sorted from total blood from a healthy subject. In order to determine the miRNA expression profile of these cells, RNA was extracted, labeled and hybridized on an Affymetrix miRNA array.
Project description:Gene expression microarray profiling was performed on peripheral blood leukocyte subsets (CD4+ T cells, CD8+ T cells, CD14+ monocytes, CD16+ neutrophils, CD19+ B cells) from healthy controls, patients with flaring autoimmune disease, and in patients with autoimmune disease following treatment, either 0 months (i.e. pre-treatment), or 3 or 12 months (into treatment).
Project description:Objective. Meningococcal sepsis remains an important cause of childhood morbidity and mortality. Largely due to logistic complexities of research in young children with acute life-threatening disease, very little is known regarding differential expression kinetics and molecular regulation of immune response genes in leukocyte subsets. Materials and methods. In this prospective case-control study, six children with meningococcal sepsis were included. Blood was drawn at four time points (t=0, t=8, t=24 and t=72 h after admission to the paediatric intensive care unit). Blood was also collected from matched controls. Detailed immunophenotyping of leukocytes was performed; RNA isolated from whole blood, lymphocytes, monocytes, and granulocytes was used to perform Affymetrix micro-array gene expression analysis. Results and conclusion. There were no differences in total leukocyte count between patients and controls. In contrast to previous in vitro studies we observed an unexpected decrease in NK cell numbers, as well as downregulation of NK cell specific and cytotoxic T-cell related gene expression in patients with meningococcal septic shock. By contrast, expression of genes, involved in innate immunity and several other pathways, differed between the different leukocyte subpopulations in a dynamic fashion. Compared to previously reported gene expression profiles, it was possible to define a meningococcal sepsis specific expression profile.
Project description:The objective of the study was to utilize DNA methylation to quantify human leukocyte subsets in human blood. This file contains data from an Illumina Infinium HumanMethlation450 for human whole blood samples as well as complex mixtures of DNA from purified human leukocyte subtypes in quantities that mimick human blood under different clinical conditions.