Bulk RNA sequencing of CD8a+ and CD8a- CD56bright and CD56dim human NK cells

In patients treated with memory-like NK cellular therapy with AML, a negative association with CD8a expression on donor NK cells and treatment response was found. The goal of this project was to determine how this finding extended to conventional NK cells, and to characterize the expression and functional role of CD8a. Peripheral blood NK cells were sorted on CD8 expression and analyzed by bulk RNA sequencing within CD56bright and CD56dim NK cell subsets. Freshly isolated peripheral blood NK cells were rested overnight in 1 ng/mL IL-15, then gated based on expression of CD56 and CD16 to distinguish CD56bright and CD56dim NK cells, then sorted into CD56bright CD8+, CD56bright CD8-, CD56dim CD8+, or CD56dim CD8- populations. Cells were placed in trizol immediately after sorting. NK cells were sorted from 6 unique healthy donors. A dump gate was used to exclude cells expressing CD3, CD19, or CD14. Comparative gene analysis was performed on CD8+ vs CD8- CD56bright and CD56dim NK cells.