Transcriptional profiling of the tuberculin skin test in individuals cured of active tuberculosis disease (RNA-Seq)
Ontology highlight
ABSTRACT: Whole genome transcriptional profiling of punch biopsies from the site of 2U intradermal tuberculin or equivalent volume of saline injection at 48 hours after injection in individuals that have been cured of active tuberculosis disease.
Project description:Whole genome transcriptional profiling of punch biopsies from the site of 2U intradermal tuberculin or equivalent volume of saline injection at 48 hours after injection in patients that have been cured of active tuberculosis (TB) disease.
Project description:Whole genome transcriptional profiling of punch biopsies from the site of 2U intradermal tuberculin or equivalent volume of saline injection at 48 hours after injection.
Project description:Transcriptome at the site of a 48 hour tuberculin skin test (TST) and saline injection from patients with active TB disease and latent TB infection
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: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:Psoriasis is a chronic, debilitating, immune-mediated inflammatory skin disease. As IFN-gamma is involved in many cellular processes, including activation of T cells and dendritic cells (DCs), antigen processing and presentation, cell adhesion and trafficking, and cytokine and chemokine production, IFN-gamma-producing Th1 cells were proposed to be integral to the pathogenesis of psoriasis. Recently, IFN-gamma was shown to enhance IL-23 and IL-1 production by DCs and subsequently induce Th17 cells, important contributors to the inflammatory cascade in psoriasis lesions. To determine if IFN-gamma indeed induces the pathways leading to the development of psoriasis lesions, a single intradermal injection of IFN-gamma was administered to an area of clinically normal, non-lesional skin of psoriasis patients and biopsies were collected 24 hours later. Although there were no visible changes in the skin, IFN-gamma induced molecular and histological features characteristic of psoriasis lesions. IFN-gamma increased a number of differentially expressed genes in the skin, including many chemokines concomitant with an influx of T cells and inflammatory DCs. Furthermore, inflammatory DC products TNF, iNOS, IL-23, and TRAIL were present in IFN-gamma-treated skin. Thus, IFN-gamma, which is significantly elevated in non-lesional skin compared to healthy skin, appears to be a key pathogenic cytokine that can induce the inflammatory cascade in psoriasis. RNA was isolated from whole skin punch biopsies of either healthy or non-lesional psoraisis patients at baseline or 24 hours after placebo or IFN-g injection.
Project description:Vitamin D insufficiency may exacerbate non-specific inflammation observed in older adults. Here, we tested the hypothesis that an inflammatory gene signature present in old skin following saline injection (as model for non-specific needle injury) normalizes after oral vitamin D3 supplementation. To define the saline-induced signature, we compared gene expression in skin biopsies taken six hours after saline injection in old adults (≥65 years) to biopsies from unmanipulated skin. We then assessed signature expression in saline-injected skin of old and young adults (<40 years), and in paired samples of old adults before and after oral vitamin D3 supplementation (6400 IU/day for 14 weeks), where median serum 25-hydroxyvitamin D increased from 43 nmol/L (interquartile range 36-53 nmol/L) to 131 nmol/L (interquartile range 115-147 nmol/L). This submission comprises 112 samples from 57 individuals.
Project description:DNA vaccines delivered with electroporation have shown promising results in pre-clinical models and are evaluated in clinical trials. Here, we aim to characterize early mechanisms occuring in the skin after intradermal injection and electroporation of the auxoGTUmultiSIV DNA vaccine in non-human primates. Firstly, we show that electroporation acts as an adjuvant by enhancing local inflammation, notably via granulocytes, monocytes/macrophages and a CD1aint -expressing cell recruitment. Electroporation also induced Langerhans cell maturation, illustrated by CD86, CD83, and HLA-DR up-regulation, and their migration out of the epidermis. Secondly, we demonstrate the crucial role of the DNA vaccine in the soluble factors release, such as MCP-1 or IL-15. Transcriptomic analysis showed that electroporation played a major role in gene expression changes post-vaccination. However, the DNA vaccine is required to strongly up-regulate several genes involved in inflammatory responses (e.g Saa4), cell migration (e.g Ccl3, Ccl5 or Cxcl10), APC activation (e.g Cd86) and interferon-inducible genes (e.g Ifit3, Ifit5, Irf7, Isg15, orMx1), illustrating an antiviral response signature. Also, AIM-2, a cytosolic DNA sensor, appeared to be strongly up-regulated, only in the presence of the DNA vaccine and trends to positively correlate with several interferon inducible genes, suggesting the potential role of AIM-2 in the vaccine sensing and the subsequent innate response activation leading to strong adaptive T-cell responses. Overall, these results demonstrate that a combined stimulation of the immune response, in which electroporation and the auxoGTUmultiSIV vaccine triggered different components of the innate immunity, led to strong and persistent cellular recall responses.
Project description:The study aimed at defining the local and systemic innate responses after intradermal injection of flagellin. For this purpose, mice were immunized by intradermal route with flagellin. Skin and blood were then sampled at selected times for microarray analyses. Mice were then immunized with the TLR5 agonist flagellin by intradermal route. Blood samples and punch biopsies of skin were sampled post-immunization and on mock animals. Total RNA was extracted and microarray experiments were performed as single-color hybridizations on Agilent 4x44K mouse whole genome catalog arrays (Agilent-014868) according supplier's recommendations.