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

嗜碱性粒细胞（Basophils）是最为罕见的粒细胞类群，仅占外周血白细胞总数的1%以下。近年来，针对嗜碱性粒细胞的分析工具取得长足进展，使我们得以明确其在过敏反应及抗寄生虫感染防护中的关键作用。尽管如此，嗜碱性粒细胞的分化轨迹仍未阐明。我们在一系列实验（包括已上传至GSE206589基因表达数据库的单细胞RNA测序（scRNA-seq）分析）中，鉴定出了此前未被认知的前嗜碱性粒细胞亚群，该亚群涵盖了此前定义的嗜碱性粒细胞前体（basophil precursors, BaPs）。本研究探究了经抗原/免疫球蛋白E（IgE）刺激或白细胞介素3（IL-3）刺激激活的前嗜碱性粒细胞与成熟嗜碱性粒细胞的基因表达谱。研究结果显示，经IL-3激活的前嗜碱性粒细胞，其基因表达谱与经抗原/IgE或IL-3刺激的成熟嗜碱性粒细胞存在显著差异，提示前嗜碱性粒细胞具有独特的生物学特性。整体实验设计：实验前1天，小鼠经静脉注射TNP特异性IgE进行致敏。从经TNP-IgE致敏的小鼠骨髓中分离得到CD34-CLEC12Ahi前嗜碱性粒细胞与CD34-CLEC12Alo成熟嗜碱性粒细胞，分别于体外在卵清蛋白（OVA，对照组）、TNP偶联卵清蛋白（TNP-OVA）或IL-3存在的条件下培养4小时，随后进行批量RNA测序（bulk RNA-Seq）分析。