Differentiation fate of a stem-like CD4 T cell controls immunity to cancer

The goals of this study were to gain a better understanding of the heterogeneity and transcriptional profile of bulk PD1+CD45RA- CD4 T cells in kidney cancer primary tumors and activated PD1+ CD4 and CD8 T cells from tumor draining lymph nodes of TRAMPC1-GP bearing mice before and after Treg depletion. Overall design: Activated CD4 T cells (live CD3+ CD19- CD8- CD4+PD1+CD45RA-) from 2 primary kidney cancer patients were FACS-purified. Additionally Activated CD4 T cells (live CD3+ CD19- B220- MHCII-CD8-CD4+) from tumor draining lymph nodes of 5-week TRAMPC1-GP bearing mice before and 5-days after Treg depletion were FACS purified. Naive CD4 T cells (PD1-CD44-CD62L+) were FACS purified as a control and spiked in from every sample. Single-cell suspensions of FACS-purified cells were loaded onto the 10X Genomics Chromium Controller. Library construction was performed using the Chromium Single Cell 5' Library Construction Kit followed by sequencing on a HiSeq3000.