Project description:The aim was to assess miRNA expression in 3 human ex-vivo CD8+ T cell subsets which span from antigen inexperienced cells (NaM-CM-/ve) to early memory cells (central memory, Tcm) and later stage memory cells (effector memory, Tem) CD8+ T cells were sorted on a FACS Aria II machine. N = naM-CM-/ve = CD8+, CCR7+, CD45RA+, CD45RO-, Tcm = central memory = CD8+, CCR7+, CD45RA-, CD45RO-,Tem= effector memory = CD8+, CCR7-, CD45RA-, CD45RO+ PBMC were isolated from 3 healthy human donors and sorted by FACS into 3 CD8+ T cell subsets. Total RNA was purified using the miRVANA kit (Ambion)
Project description:To investigate the epigenetic landscape of tumor-reactive exhausted CD8+ T cells, ATAC-seq was performed on fluorescence-activated cell sorted CD8+ T cell subsets isolated from Lag3^iCreERT2^ Rosa26^LSL-tdTomato^ mice bearing B16-F10 melanoma tumors. Chromatin accessibility profiles were generated for lineage-traced LAG3^+tdTomato^+, LAG3^-tdTomato^+, and corresponding control CD8+ T cell populations isolated from tumors and draining lymphoid tissues. These data were used to characterize chromatin accessibility changes associated with exhausted, progenitor-like, and memory-like CD8+ T cell states and to define epigenetic relationships among lineage-traced T cell populations.
