RNA-seq of CD4+ T cell subsets in LCMV infection. RNA-seq of CD4+ T cell subsets in LCMV infection

T follicular helper (TFH) cells are a distinct type of CD4 T cells that are essential for most antibody and B lymphocyte responses. TFH regulation and dysregulation is involved in a range of diseases. Bcl6 is the lineage defining transcription factor of TFH cells and its activity is essential for TFH cell differentiation and function. However, how Bcl6 controls TFH biology has largely remained unclear, at least in part due to intrinsic challenges of connecting repressors to gene upregulation in complex cell types with multiple possible differentiation fates. Multiple competing models were tested here by a series of experimental approaches to determine that Bcl6 exhibits negative autoregulation and controls pleiotropic attributes of TFH differentiation and function (migration, costimulation, inhibitory receptors, and cytokines) via multiple repressor-of-repressor gene circuits. Overall design: 7 samples, 3 replicates each, naïve or Day 7 LCMV infection, SMARTA CD4 T cell subsets

滤泡辅助性T（T follicular helper, TFH）细胞是一类独特的CD4阳性T细胞，在绝大多数抗体应答与B淋巴细胞应答过程中发挥关键作用。TFH细胞的稳态调控与功能失调均与多种疾病的发生发展密切相关。Bcl6是TFH细胞的谱系决定性转录因子，其活性对TFH细胞的分化与功能维持不可或缺。然而，Bcl6调控TFH细胞生物学特性的具体机制仍未明确，这在一定程度上源于在具备多种分化潜能的复杂细胞类型中，将转录抑制因子与基因上调事件关联起来所面临的内在挑战。本研究通过一系列实验手段对多种竞争性模型进行验证，最终证实Bcl6存在负自调控，并通过多条阻遏因子级联基因回路（repressor-of-repressor gene circuits）调控TFH细胞分化与功能的多效性特征，涵盖细胞迁移、共刺激、抑制性受体及细胞因子相关通路。实验设计概述：共包含7个样本，每个样本设置3次生物学重复，分组为未致敏（naïve）小鼠，以及感染淋巴细胞性脉络丛脑膜炎病毒（Lymphocytic choriomeningitis virus, LCMV）第7天的小鼠，检测对象为SMARTA CD4阳性T细胞亚群。