Project description:Proteome characterization of isolated mitochondria samples prepared from three commonly used acute leukemia cell lines (HL-60, KG-1, MV-4-11). Data were compared to isolated mitochondria prepared from peripheral blood mononuclear cells (PBMC), isolated from healthy human subjects.
Project description:Whole blood was collected from healthy and autistic infants and peripheral blood mononuclear cells (PBMC) were isolated. Transcriptional profile in PBMCs was compared between healthy and autistic infants.
Project description:Whole blood was collected from healthy adult and peripheral blood mononuclear cells (PBMC) were isolated. PBMCs were cultured in medium or stimulated with RNase treated EC-12 or untreated EC-12 for 6hours. Then transcriptome profiles in PBMC were obtained.
Project description:Context
Glucocorticoids used in pharmacological doses for the treatment of a variety of medical conditions, and endogenous glucocorticoid excess – Cushing’s syndrome, may result in several adverse effects, but currently there is no clinically useful biomarker of glucocorticoid activity.
Objective
To identify glucocorticoid-responsive proteins potentially measurable in human serum.
Setting and Design
A three-phase protein biomarker discovery strategy was used. Proteomic biomarker discovery and qualification was conducted on the secretome of ex vivo-stimulated peripheral blood mononuclear cells (PBMC) isolated from 6 volunteers, incubated ± dexamethasone 100 ng/mL for 4h and 24h. Untargeted proteomics with label-free quantification (LFQ) was conducted to discover candidate proteins which were quantified using targeted proteomics by a custom multiple reaction monitoring mass spectrometry (MRM-MS) assay. Five proteins were selected for serum measurement by immunoassay in 20 healthy volunteers, with blood drawn at baseline and 12h after 4 mg oral dexamethasone.
Project description:We performed systems analyses of immune responses to the meningococcal polysaccharide (MPSV4) and conjugate (MCV4) vaccines in healthy adults. The goal was to identify innate gene signatures that correlate to antibody responses induced by these vaccines. Healthy adult volunteers (18-45 years of age) were randomized to be vaccinated subcutaneously with MPSV4 (Menomune®, n=13) or intramuscularly with MCV4 (MenactraTM, n=17) (Sanofi Pasteur, Swiftwater, PA). Whole blood samples were collected using CPT tubes at days 0, 3 and 7 post-vaccination. Peripheral blood mononuclear cells (PBMC) were isolated from fresh blood and used for DNA microarray experiments.
Project description:Aberrant gene expression analysis between peripheral blood mononuclear cell (PBMC) samples from healthy individuals and patients with pancreatic carcinoma, gastric carcinoma and hepatocellular carcinoma (HCC) were identified using Affymetrix gene arrays. Peripheral blood mononuclear cell (PBMC) from healthy individuals, patients with pancreatic carcinoma, gastric carcinoma and HCC were isolated and total RNA was extracted for Affymetrix gene microarray analysis.
Project description:The long-term effects of neonatal intermittent hypoxia (IH), an accepted model of apnea-induced hypoxia, are unclear. We have previously shown lasting âprogrammingâ effects on the HPA axis in adult rats exposed to neonatal IH. We hypothesized that neonatal rat exposure to IH will subsequently result in a heightened inflammatory state in the adult. Rat pups were exposed to normoxia (control) or six cycles of 5% IH or 10% IH over one hour daily from postnatal day 2 â 6. Plasma samples from blood obtained at 114 days of age were analyzed by assessing the capacity to induce transcription in a healthy peripheral blood mononuclear cell (PBMC) population and read using a high-density microarray. The analysis of plasma from adult rats previously exposed to neonatal 5% IH vs. 10% IH resulted in 2,579 significantly regulated genes including increased expression of Cxcl1, Cxcl2, Ccl3, Il1a, and Il1b. We conclude that neonatal exposure to intermittent hypoxia elicits a long-lasting programming effect in the adult resulting in an upregulation of inflammatory-related genes. Apnea is the most common cause of neonatal hypoxia affecting about 50% of preterm births (30 â 31 weeks), usually due to immature respiratory development. Upregulation of inflammatory genes and pathways in children 7 â 10 years of age has been shown, and there is a known increased risk of insulin resistance in adulthood when the fetus is exposed to maternal hypoxia, but the mechanism is unclear. The long-term metabolic, endocrine, and immunological effects of neonatal intermittent hypoxia (IH) exposure, an accepted model of apnea-induced hypoxia, have not been thoroughly evaluated. Recent studies in rats have shown that perinatal IH exposure can result in oxidative stress, causing a permanent immune response subsequently resulting in features of diabetes mellitus. We have previously examined adult rats exposed to neonatal intermittent hypoxia and perinatal continuous hypoxia, and have found lasting âÂÂprogrammingâ effects on the HPA axis. We now assess the long term effects of an accepted model of apnea-induced hypoxia using a validated transcriptional bioassay to study the extracellular milieu of adult rats exposed to neonatal intermittent hypoxia. We hypothesize that exposure to neonatal intermittent hypoxia will result in an increased inflammatory state in the adult as a result of long-lasting programming. Sprague-Dawley (SD) rat pups were treated with neonatal normoxia (21% O2, control), 5% intermittent hypoxia (IH), or 10% IH on postnatal days (PD) 2-6, daily over 1 hr. They were reared normally by birth dams and weaned at PD22. Males were allowed to mature and sacrificed at age PD114 after an overnight fast. Whole blood collected by decapitation into tubes with EDTA, and plasma saved for further analysis. Two adult (~180 day) male Brown Norway (BN) rats served as PBMC donors. Cells were incubated with 20% plasma that was either autologous BN (self-control), or one of 3 pools: a) SD normoxic N=8, b) SD 5% IH treated N=5, and c) SD 10% IH N=3.
Project description:Aberrant gene expression analysis between peripheral blood mononuclear cell (PBMC) samples from healthy individuals and patients with chronic hepatitis B carriers and HCC were identified using Affymetrix gene arrays.M-BM- Peripheral blood mononuclear cell (PBMC) from healthy individuals, patients with patients with chronic hepatitis B carriers and HCC were isolated and total RNA was extracted for Affymetrix gene microarray analysis.
Project description:With this experiment, we aimed at showing changes in the proteome and secretome of porcine peripheral blood mononuclear cells (PBMC) after stimulation with Banana Lectin (BanLec).