RNA-sequencing of Aire-ILC3 cells from wild-type and Aire-deficient mice

Autoimmune regulator (Aire) plays an indispensable role in the induction of central immune tolerance. We identify a novel subset of Rorγt-dependent innate lymphoid cell-type in secondary lymphoid organs, which presents endogenously generated antigens and contributes to peripheral T cell tolerance. In this experiment, transcriptomes of these Aire-ILC3 cells are characterized in wild-type Balb/C and Aire-/- Balb/C mice [Ramsey, C. et al. (2002) Hum Mol Genet 11:397-409] using RNA-seq method. To obtain the cells, peripheral lymph nodes were minced, enzymatically digested, and Aire-ILC3 cells were FACS-sorted as Lin-, MHCII+, CD80+, IL7Ra+ cells.自身免疫调节因子（Autoimmune regulator, Aire）在中枢免疫耐受的诱导过程中发挥不可或缺的作用。本研究在次级淋巴器官中鉴定出一种新型的维甲酸相关孤儿受体γt（Rorγt）依赖的先天性淋巴细胞亚型，该细胞可呈递内源性抗原，并参与外周T细胞耐受的调控。本实验采用RNA测序（RNA-seq）技术，对野生型Balb/C小鼠及Aire基因敲除（Aire-/-）Balb/C小鼠体内的Aire-ILC3细胞的转录组进行特征分析[Ramsey C 等，2002，Hum Mol Genet，11:397-409]。具体细胞获取流程如下：对外周淋巴结进行剪碎处理并实施酶解，随后通过荧光激活细胞分选术（FACS）按照Lin阴性、MHCII阳性、CD80阳性、IL7Ra阳性的表型分选得到Aire-ILC3细胞。