How do CD8+ T cells complete DNA replication in less than 4 hours?

Abstract

Abstract We demonstrate that activated CD8+ T cells from Pmel mice replicate their genome within 4 hours which is significantly faster than most cancer cells. Many genotoxic chemotherapeutic agents target the rapid proliferation of cancer cells. However, activated T cells also expand rapidly and this can lead to lymphopenia in patients receiving these chemotherapeutic agents. Understanding how CD8+ T cells complete DNA replication in less than 4 hours could lead to the development of treatment schedules that spare CD8+ T cells. Three possible mechanisms may underlie the rapid DNA replication in CD8+ T cells: 1) faster replication fork velocity, 2) higher density of origin firing, or 3) modified replication timing program, allowing for simultaneous replication of early- and late-replicating regions of the genome. Using DNA combing, we show that the replication fork velocity and inter-origin distance is similar in unperturbed CD8+ T cells and B16 cancer cells. Repli-seq analysis, a NGS genome-wide replication timing analysis, suggest that regions of the genome replicated late in S phase in fibroblasts are replicated earlier in CD8+ T cells, suggesting that the rapid DNA replication in CD8+ T cells is a consequence of a modified, truncated replication timing program. We also show for the first time that actively replicating CD8+ T cells have dormant origins suggesting that this mechanism of genome stability is maintained in activated CD8+ T cells.