Allergen-stimulated CD4+ T cells from hen's egg allergic patients

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 PBMCs from three hen’s egg allergic children and two non-allergic children were cultured in RPMI 1640 supplemented with 10% autologous plasma for 16 hours with or without hen’s 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 sorter（Miltenyi Biotec, Bergisch Gladbach, Germany）after 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 manufacturer’s instruction.