Regulation of CD8 T cell differentiation by the RNA-binding protein DDX5

The RNA-binding protein DDX5 is a polyfunctional regulator of gene expression, but its role in CD8+ T cell biology has not been extensively investigated. In this study, we demonstrate that deletion of DDX5 reduced differentiation of terminal effector (TE), effector memory (TEM), and terminal effector memory (t-TEM) cells while increasing the generation of central memory (TCM) cells; forced expression of DDX5 elicited the opposite phenotype. DDX5-deficient CD8+ T cells exhibited increased expression of genes that promote TCM cell differentiation, including Tcf7 and Eomes. Together, these findings reveal a role for DDX5 in regulating the differentiation of effector and memory CD8+ T cell subsets in response to microbial infection. Overall design: WT CD45.1 and Ddx5fl/flCd4-Cre+ CD8+ CD45.1.2 P14 cells were adoptively co-transferred into congenically distinct recipient mice. The recipient mice were infected with 2×105 PFU LCMV 24 hours after transfer and sacrificed on days 4, 7, and 30 post-infection. Splenic P14 T cells were isolated by FACS. About 10,000 cells per sample were loaded into Single Cell G chips (10x Genomics) and partitioned into Gel Bead In-Emulsions in a Chromium Controller (10x Genomics). Single-cell RNA libraries were prepared according to the 10x Genomics Chromium Next GEM Single Cell 3' v3.1 (Dual Index) User Guide and sequenced on a HiSeq 4000 (Illumina).