Project description:We measured TNFα activity on the transcriptional level by identifying TNFα-inducible genes in human blood monocyte-derived macrophages (MDM). On the basis that TNFα does not act in isolation, we used bacterial lipopolysaccharide (LPS) as a prototypic model of a stimulus to invoke secretion of a wide range of immunologically active factors, including TNFα. In order to identify gene expression attributable to LPS-induced TNFα, we compared the transcriptome of MDM 24 hours after stimulation with 100 ng/ml ultra-pure LPS in the presence and absence of the soluble TNF receptor fusion protein Etanercept (10 µg/ml).
Project description:Vitamin D is widely reported to inhibit innate immune signalling and dendritic cell (DC) maturation, leading to attenuation of DC mediated T cell activation as a potential immunoregulatory mechanism to guard against autoimmunity and immmunopathology. It is not known whether vitamin D has global or gene specific effects on transcriptional responses downstream of innate immune stimulation. In order to address this question, we used genome-wide transcriptional profiling of monocyte derived DC differentiated in the presence and absence of vitamin D, and then stimulated with and without lipopolysaccharide for four hours.
Project description:Anti-TNF therapy has transformed the management of rheumatoid arthritis (RA); however little is known about its effects on cell mediated immune responses and TNF-inducible activity at the site of inflammation. Here, we used the tuberculin skin test (TST) as a standardised in vivo human challenge model to make systems level assessments of the effects of anti-TNF therapy in a prototypic cell mediated immune response. RA patients (treated with monoclonal anti-TNF antibodies (adalimumab or infliximab), the soluble TNF receptor etanercept or the standard therapy methotrexate) and healthy volunteers with immunological memory to Mycobacterium tuberculosis antigens were identified using an interferon-γ release assay of peripheral blood. Two units tuberculin or an equivalent volume of saline was injected into the forearm of study participants (n=50). After 72 hours, 3 mm skin punch biopsies were taken from the injection site and processed for whole genome transcriptional profiling by Agilent gene microarray.
Project description:Whole blood transcriptional profiles of patients with (1) active pulmonary ['AdjuVIT active TB' and 'New active pulmonary'] and (2) extrapulmonary TB ['New active-extrapulmonary'] at time of diagnosis, (3) long-term recovery after treatment for active pulmonary TB ['AdjuVIT active TB'], (4) febrile illnesses presenting to hospital ['Fever mixed infection'] and (5) febrile pneumonia ['Fever pneumonia'] before antibiotic treatment, and (6) healthy vounteers. Each array sample represents a separate individual in each group. This submission includes two human whole genome Agilent Array Designs: A-MEXP-2104 and A-AGIL-28. Each of the individual raw array files are included as well as a single processed file representing the data matrix of all the merged and normalised data for the probes that are shared by the two array designs.
Project description:Prolonged rupture of membranes (PROM) is thought to incur higher risk of neonatal infection, leading to expedited delivery and/or the use of empirical perinatal antibiotics. Here, we compared the transcriptional profile of neonatal cord blood between babies where rupture of membranes occurred greater than 24 hours before delivery (= PROM cases) and babies where rupture of membranes occurred less than 24 hours before delivery (= Control cases). On the basis that perinatal infection is more likely in births associated with PROM than those without, we tested the hypothesis that perinatal infection in a subset of births following PROM, exhibit immune responses evident in the neonatal cord blood transcriptome.
Project description:We tested the hypothesis that anti-tumor necrosis factor (TNF) therapy reduces TNF-inducible gene expression in blood. Whole peripheral blood from healthy volunteers and rheumatoid arthritis patients (treated with the monoclonal anti-TNF antibody adalimumab, the soluble TNF receptor etanercept or the standard therapy methotrexate) was incubated at 37 °C for three hours in the presence or absence of 10 ng/ml recombinant human TNF. Blood samples were then processed for RNA isolation and transcriptional profiling by gene microarray. This submission includes two human whole genome Agilent Array Designs: A-MEXP-2104 and A-GEOD-20844. Each of the individual raw array files are included as well as a single processed file representing the data matrix of all the merged and normalised data for the probes that are shared by the two array designs.