Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T-cells [ATAC-Seq]

Since exhausted CD8 T-cells are known to have a strong epigenetic imprint, which is a major obstacle for reinvigoration, we investigated whether the induced Klf4 expression could change the epigenetic status of the exhausted CD8 T-cells. For this, we performed ATAC-sequencing and compared chromatin landscapes between naïve, in vitro generated effector, exhausted (GFP-induced), and reinvigorated (KLF4-induced) CD8 T-cells. Overall design: ATAC-sequencing for naïve, in vitro generated effector, exhausted (pRetroX-GFP/Dox+), and reinvigorated (pRetroX-Klf4/Dox+) CD8 T-cells.

鉴于耗竭型CD8 T细胞（exhausted CD8 T-cells）已被证实具有较强的表观遗传印记（epigenetic imprint），这是其功能重振的主要障碍，本研究旨在探究诱导型Krüppel样因子4（Klf4）表达能否改变耗竭型CD8 T细胞的表观遗传状态。为此，我们开展了转座酶可及性测序（ATAC-sequencing），并对比了初始型CD8 T细胞（naïve）、体外诱导的效应型CD8 T细胞、绿色荧光蛋白（GFP）诱导的耗竭型CD8 T细胞以及KLF4诱导的功能重振型CD8 T细胞的染色质开放景观。整体实验设计：针对初始型CD8 T细胞、体外诱导的效应型CD8 T细胞、pRetroX-GFP/Dox+诱导的耗竭型CD8 T细胞以及pRetroX-Klf4/Dox+诱导的功能重振型CD8 T细胞进行转座酶可及性测序。