Gene expression profiles of pre-basophils versus mature basophils stimulated with antigen/IgE- or IL-3-stimulation.

Basophils are the least common granulocytes which represent less than 1% of peripheral blood leukocytes. Recent development of analytical tools for basophils enables us to understand their critical roles in allergic reactions and protection from parasitic infections. Nevertheless, the differentiation trajectory of basophils remains unclear. We have identified previously-unappreciated pre-basophil subpopulation which encompasses previously-defined basophil precursors (BaPs) in a series of experiments including scRNA-seq analysis (deposited to GSE206589). In this study, we explored gene expression profiles of pre-basophils and mature basophils activated by antigen/IgE-stimulation or IL-3-stimulation. We have revealed that IL-3 activated pre-basophils displayed distinct gene expression profiles from antigen/IgE- or IL-3-stimulated mature basophils, suggesting the unuque property of pre-basophils. Mice were intravenously sensitized with TNP-specific IgE 1 day prior to the experiment. CD34-CLEC12Ahi pre-basophils and CD34-CLEC12Alo mature basophils isolated from the bone marrow of TNP-IgE sensitized mice were separately cultured ex vivo in the presence of OVA (Control), TNP-conjugated OVA (TNP-OVA), or IL-3 for 4h and were subjected to bulk RNA-Seq analysis.