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: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:Transcriptional profiling of biopsies from tuberculin skin test sites was performed to investigate the interaction of complex cellular and molecular networks at the interface between innate and adaptive immunity which coordinate anti-mycobacterial responses in vivo.
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.