Project description:Allergen-stimulated T cells from henâs egg-allergic children were analyzed to identify genes that are specifically up-regulated in these cells. Experiment Overall Design: PBMCs from three henâs egg allergic children and two non-allergic children were cultured in RPMI 1640 supplemented with 10% autologous plasma for 16 hours with or without henâs egg white allergen (allergen concentration10 mg/ml). To focus on specific T cells reacting with HEW, cells bearing CD4 antigen were isolated from cultured PBMCs using a Magnetic cell sorterï¼Miltenyi Biotec, Bergisch Gladbach, Germanyï¼after CD14 positive cell depletion. Total RNA was extracted from these CD4 positive cells with an RNeasy mini-kit (Qiagen, Valencia, CA, USA) according to the manufacturerâs instruction.
Project description:Allergen-stimulated T cells from hen’s egg-allergic children were analyzed to identify genes that are specifically up-regulated in these cells. Keywords: disease state analysis
Project description:Gene expression (Npatients = 21, Ncontrols = 21) of CD4+ T-cells failed to seperate patients with seasonal allergic rhinitis (SAR) and healthy controls in an in vitro model system in which purified PBMCs from patients and healthy controls were challenged with allergen for 7 days. PBMCs from 21 patients (P) and 21 healthy controls (H) were challenged with grass pollen for 7 days. Diluent challenged control samples were obtained from all subjects. CD4+ cells were purified by MACS.
Project description:Six patients with seasonal allergic rhinitis were challenged daily for 8 days with birch pollen extract. A mucosal biopsy was obtained from one nostril at basline (day 0) and from the other nostril after allergen challenge (day 9). The mucosal biopsies were digested into single cells, and then sorted into CD4 T cells and CD45+HLA-DR+ cells. Total RNA was extracted, amplified using whole transcriptome amplification, and gene expression was profiled on microarrays. The study design consisted of 6 subjects, 2 cell types (CD4 T cells, CD45+ HLA-DR+ cells), and 2 conditions (baseline, allergen challenge).
Project description:Gene expression (Npatients = 21, Ncontrols = 21) of CD4+ T-cells failed to seperate patients with seasonal allergic rhinitis (SAR) and healthy controls in an in vitro model system in which purified PBMCs from patients and healthy controls were challenged with allergen for 7 days.
Project description:Gene expression analysis in CD4+ T cells extracted from allergen-challenged PBMCs, isolated from discordant MZ twins with IAR MZ twins discordant for intermittent allergic rhinitis (IAR)
Project description:Seasonal allergic rhinitis (SAR) is a complex disease that is caused by many interacting genes and environmental factors. It is also an excellent model disease for clinical studies; it is common, it is seasonal, and since it takes place in the nasal cavity it can be studied in vivo non-invasively. Furthermore, the key disease cell, the Th2 cell is known. We study SAR using allergen-challenged CD4+ cells from allergic patients.
Project description:The transcriptional response to egg differed between PBMCs from egg allergic and clinically tolerant subjects. Differentially expressed genes included IL-9 and TNFg. Enrichment analysis of differentially expressed gene signatures and associated co-expressed gene modules revealed an association of egg allergy with unique immune pathways. In addition to showing a positive association of egg-induced gene transcription in allergic individuals with Th2 CD4+ T cells and a novel negative association with induced Tregs, this approach identified a highly significant overlap with genes induced by TLR4 stimulation of myeloid cells.
