Identification of novel activation marker and regulator of human basophils via transcriptomic analysis of IL-3-stimulated basophils

Human basophils regulate allergic immune responses by releasing histamine and effector cytokines such as interleukin 4 (IL-4) and IL-13. IL-3/IL-3R signaling axis plays key roles in the maturation, survival and activation of basophils. We and others have shown that IL-3-induced surface receptors e.g. ST2 and IL-17RB regulate the biology of basophils. We in this study aim to identify new basophil activation maker and regulator by transcriptomic analysis of IL-3-stimulated basophils. Human basophils were pre-enriched from PBMCs from buffy coat by Magnetic-activated cell sorting (MACS) positive sorting using rat anti-human CRTH2 microbeads. Next, basophils were further purified by FACS sorting of FSClowSSClowCD4-CRTH2high populations.Purified basophils at a density of 2×105 cells/well in a 96-well plate were cultured either with or without human IL-3 (50 ng/mL) (n = 3) for 24 hours. Gene expression microarray analysis was performed using SurePrint G3 Human Gene Expression 8×60K V2 microarray according to the manufacturer’s instruction.