RNA-Seq analysis of P14 CD8+T cells in B16-GP tumors

CD8+ T cells play an important role in fighting against tumors. However, CD8+ T cells often become functionally impaired or “exhausted” in the tumor microenvironment. T cell exhaustion is a major hurdle that currently limits the efficacy of immunotherapeutic regimens. The exhausted T cell population is heterogeneous and contains T cells subsets at varying states of differentiation and with a range of effector functions and proliferative potential. Identification of exhausted T cell-specific markers will help to understand the heterogeneity of this population. Therefore, we have performed RNA-sequencing analysis of mouse tumor-infiltrating CD8+ T cells and splenic CD8+ T cells. We implanted C57BL/6 mice with B16-GP33-41 melanoma cells. On day 11 after tumor cell implantation, TCR-transgenic P14 CD8+ T cells were adoptively transferred to the tumor-bearing mice. The GP33-41 epitope is recognized by the P14 T cells. The tumor-infiltrating P14 CD8+ T cells were FACS-sorted from tumor and spleen for RNA preparation using the Qiagen RNeasy Mini Kit. The DKFZ High Throughput Sequencing Unit prepared and sequenced the library (Illumina HiSeq 2000 v4 Single-Read 50 bp).