Identifying key transcriptional regulators that establish T cell memory during viral infection

Abstract

During infections, naive CD8 T cells differentiate into terminal effector cells (TE) that are relatively short-lived, and memory precursor (MP) cells that give rise to long-lived memory CD8 T cells, but the transcriptional control of this process is still unclear. In naïve CD8 T cells, cis-regulatory regions that become accessible in chromatin during the first 24 hours of TCR stimulation and that remain accessible in mature memory T cell subsets are highly enriched with motifs encoding binding sites for the ETS- and bZIP-families of transcription factors (TFs), many of which also overlap Runx-TF binding sites. The ETS and bZIP TF families are encoded by 88 genes, and are differentially expressed between naïve, early effector, and memory CD8 T cell subsets. To interrogate their requirements functionally, we applied an in vivo pooled RNA interference screen using short hairpin RNAs in microRNA contexts (shRNAmirs) to suppress individually all TFs from both families in CD8 T cells responding to lymphocytic choriomeningitis virus (LCMV) infection. In addition, we focused on Ets1, the most highly expressed ETS TF in naïve, effector and memory cell subsets. Ets1 suppression with shRNAmirs impaired overall effector CD8 T cell numbers in vivo, and increased the frequency of TE-like cells at early times after LCMV infection, which resulted in reduced long-lived effector cells and an increased fraction of altered central memory-like cells at later time points. Enforced Runx3 expression in Ets1-suppressed cells partially restored normal MP cell formation at early times. These results suggest that Ets1 and Runx3 are each necessary for the normal differentiation of MP cells, and could function cooperatively to program their differentiation.