Regulation of the alveolar regenerative niche by amphiregulin-producing Treg cells (AT2)

This study investigates the role of regulatory T cell–derived Areg in driving epithelial repair following influenza infection. We use RNA sequencing to probe the gene expression changes within epithelial cells in wildtype mice and in mice that conditionally lack Areg from T cell sources (CD4-cre Areg-flox). Further, we identify a population of damage-associated transitional alveolar epithelial cells that are induced in reponse to acute injury and exhibit gene expression changes reflective of AT2 transdifferentiation to AT1. Overall design: Alveolar epithelial cells were isolated by FACs prior to RNA extraction and RNA sequencing. Wild-type type II alveolar epithelial cells (AT2) isolated from mock treated (PBS) mice and mice treated with PR8/H1N1 influenza at day 7 post infection; n= 4 per group. Intermediate alveolar epihtelial cells isolated at day 5 post infection with PR8/H1N1 influenza from CD4-cre Areg-fl/fl and Areg-fl/fl controls; n=4 per group.

本研究探讨了调节性T细胞（regulatory T cell）来源的双调蛋白（Areg）在流感病毒感染后驱动上皮修复过程中的作用。
我们采用RNA测序（RNA sequencing）技术，检测野生型小鼠以及T细胞特异性条件性敲除双调蛋白的小鼠（CD4-cre Areg-flox）的上皮细胞内的基因表达变化。
此外，我们还鉴定出一群损伤相关的过渡态肺泡上皮细胞：该类细胞在急性损伤后被诱导产生，其基因表达特征可反映AT2向AT1的转分化过程。
实验整体设计如下：在提取RNA并进行RNA测序前，通过荧光激活细胞分选（FACS）分离肺泡上皮细胞。具体分组包括：从经磷酸盐缓冲液（PBS）模拟处理的小鼠，以及感染PR8/H1N1流感病毒后第7天的小鼠中分离野生型II型肺泡上皮细胞（AT2），每组样本量n=4；从感染PR8/H1N1流感病毒后第5天的CD4-cre Areg-fl/fl及Areg-fl/fl对照小鼠中分离中间态肺泡上皮细胞，每组样本量n=4。