Single-cell and bulk RNA-seq of Th2

Th2 cell is a subset of CD4+ T helper (Th) cells, which regulates the immunity to extracellular parasites and allergic inflammation. Th2 cells specifically secrete Type 2 cytokines such as IL-4, IL-5, IL-13, and also express many universal cytokines like IL-2 and GM-CSF. While theses cytokines are indispensable for Th2 cell mediated response, there is complex heterogeneity in the pattern of cytokine production among Th2 cell population. As previous studies were mainly conducted at population levelref, here we took advantage of single-cell RNA-seq to explore heterogeneity in in vitro differentiated Th2 cells. Although a few single cell researches about Th2 cells was conducted, there is no comprehensive understanding of the heterogeneity of Th2 lineage yet, inspiring us to make some difference on this issue. Our studies identify that cytokine genes contributed most to the transcriptome heterogeneity of Th2 cells; expression of universal cytokines showed bimodality, but not Th2 specific cytokines. Overall design: 56 Single Th2 cells for single-cell RNA-seq. Pooled 10,000 Th2 cells for bulk RNA-seq.Th2细胞是CD4+辅助性T细胞（CD4+ T helper, Th）的一个亚群，可调控胞外寄生虫免疫应答与变应性炎症反应。Th2细胞能够特异性分泌IL-4、IL-5、IL-13等2型细胞因子，同时也可表达IL-2、GM-CSF等多种泛细胞因子。尽管上述细胞因子对于Th2细胞介导的免疫应答不可或缺，但Th2细胞群体内的细胞因子产生模式存在复杂的异质性。既往研究多基于细胞群体水平开展，本研究借助单细胞RNA测序（single-cell RNA-seq）技术，对体外分化的Th2细胞的异质性进行探究。尽管已有少量针对Th2细胞的单细胞研究，但目前学界对Th2细胞谱系的异质性仍缺乏全面认知，这促使我们针对该问题展开相关研究。本研究发现，细胞因子基因对Th2细胞转录组异质性的贡献度最高；泛细胞因子的表达呈现双峰分布，而Th2细胞特异性细胞因子则无此特征。整体实验设计：选取56个单个Th2细胞进行单细胞RNA测序（single-cell RNA-seq），同时将10000个Th2细胞混合制备样本以开展批量RNA测序（bulk RNA-seq）。