The transcription factor ThPOK acts late in helper T cell lineage specification and suppresses Runx-mediated commitment to the cytotoxic T cell lineage

The transcription factor ThPOK has been shown to be required and sufficient for CD4(+)CD8(−) thymocyte generation, yet the mechanism through which ThPOK orchestrates CD4 helper T cell lineage differentiation remains unclear. Here we utilized reporter mice to track expression of transcription factors in developing thymocytes. Distal promoter-driven Runx3 (Runx3d) expression was restricted to MHC class I-selected thymocytes. In ThPOK-deficient mice, Runx3d expression was de-repressed in MHCII-selected thymocytes, contributing to their redirection to the CD8 T cell lineage. In the absence of both ThPOK and Runx, redirection was prevented and cells potentially belonging to the CD4 lineage, presumably specified independently of ThPOK, were generated. Our results suggest that MHCII-selected thymocytes are directed towards the CD4 lineage independently of ThPOK, but require ThPOK to prevent Runx-dependent differentiation towards the CD8 lineage.