Ptpn2 regulates the generation of cytotoxic Tim-3+ CD8+ T cells and restrains anti-tumor immunity

Cellular and transcriptional experiments demonstrate that deletion of Ptpn2 enhances CD8+ T cell responses to chronic viral infection and cancer. Overall design: Murine bone-marrow chimeras were generated by intravenous transfer of lineage- Sca-1+ Kit+ (LSK) cells, derived from a CD45.1+ Cas9+ expressing mouse and infected with either control or Ptpn2-targeting sgRNAs, into irradiated recipient CD45.2 mice. CD8+ cells derived from the reconstituted hematopoetic compartment of the recipient mice were then isolated for transfer to host mice. For LCMV co-transfer studies, cells were transferred (500:500 mix) to recipient mice on day -1, and mice were infected with LCMV Clone 13 on day 0. For tumor co-transfer studies, cells were transferred (1000:1000 mix) to recipient mice on day -1, and mice were injected with MC38-OVA or B16-OVA on day 0. Day 7 or 8 post tumor or virus injection respectively, co-transferred T cells were isolated from the tumor or spleen (LCMV) (as above) and replicates of 500 cells were FACS sorted for transcriptional profiling. A file with detailed methods is included at the foot of this record.