Sepsis leads to lasting changes in phenotype and function of memory CD8 T cells (RNA-Seq)

Determine genome-wide open chromatin in P14 memory CD8 T cells 30 days after septic insult Overall design: A small number of naïve P14 CD8 T cells (Thy1.1) were adoptively transferred into congenically distinct (Thy1.2) wild type hosts to generate P14 chimeras. P14 chimeric mice were then infected with LCMV-Armstrong. 30-days post infection mice underwent sham or CLP surgery. Memory P14 CD8 T cells were isolated and sorted 30-days post surgery (60-days post-infection) for analysis.检测脓毒症应激后30天的P14记忆性CD8 T细胞的全基因组开放染色质状态。 实验设计概述：将少量初始型P14 CD8 T细胞（Thy1.1）过继转移至遗传背景相异（Thy1.2）的野生型宿主小鼠体内，构建P14嵌合小鼠模型。随后对该嵌合小鼠接种淋巴细胞性脉络丛脑膜炎病毒-阿姆斯特丹株（LCMV-Armstrong）。感染后30天，对小鼠分别实施假手术或盲肠结扎穿刺（cecal ligation and puncture, CLP）手术造模。于术后30天（即感染后60天）分离并分选记忆性P14 CD8 T细胞，用于后续分析。