Extended Data Fig. 10: IFNg is intrinsically required for tumor specific CD8 LN-stem to effector differentiation.
From: Differentiation fate of a stem-like CD4 T cell controls immunity to cancer
a) Schematic to make IFNgR1 KO P14s using an HSC BM chimera lentiviral system. b-d) Phenotypic analysis of transferred VEX+ P14s in WT mice with and without doxycycline administration and Treg depleted (DTR) mice without doxycycline treatment in TDLNs and tumor. Medians are represented in each summary plot (n = 4–5 mice per group). e-f) Representative flow cytometry stain of IFNgR1 and PD1 expression in WT and KO P14s after transfer into FOXP3-DTR TRAMPC1-GP mice. Endogenous GP33 + CD8 T cells in each respective mouse are included as a comparison. Medians are represented in each summary plot from one representative experiment and analyzed by two-sided unpaired Mann Whitney U test between WT and KO (n = 7 mice). g-h) Phenotype of transferred WT and IFNgR1 KO P14s in TDLNs 5-days after Treg depletion. i) Total number and phenotypic analysis of endogenous GP33 + CD8 T cell numbers in TDLNs in mice with transferred WT and KO P14s. j) P14 number and phenotype in the tumor 5-days after Treg depletion. Data are representative of 2 independent experiments (n = 5–6 mice per group for each experiment). Median are represented in each summary plot and were analyzed by two-sided unpaired Mann-Whitney U test. k) Representative phenotype of activated (PD1 + CD45RA-) CD4 T cells in primary tumors upon surgical resection in a cohort of 47 kidney cancer patients that received immunotherapy. Patients where stratified based on the % of total TBET + PD1 + CD4 T cells in the resected primary tumors prior to therapy. l) Representative phenotype of activated (PD1 + CD45RA-) CD8 T cells in Th1 low and Th1 high patients. Medians are represented and analyzed by two-sided unpaired Mann-Whitney U test (n = 47 kidney cancer patients). m) Spearman correlation between TBET + CD4 T cells and effector GZMB + CD8 T cell populations as a percent of total cells in non-metastatic draining lymph nodes from kidney cancer patients (n = 12). n) Proposed models of T cell differentiation states in TDLNs. Restricted TDLN state. Tregs actively suppress stem-like CD4 T cells, preventing their differentiation into Th1 cells and promoting iTreg differentiation. Minimal CD4 T cell help is provided in this state and CD8 T cells are maintained in an activated stem-like state in TDLNs. Anti-tumor response is not optimal, and tumor outcompetes the T cell response. Active TDLN state. In the absence of Treg suppression stem-like CD4 T cells undergo Th1 differentiation. Th1 CD4s secrete IFNg which promotes stem-like to effector CD8 T cell differentiation. Anti-tumor effector response is optimal, and tumor is controlled. Stem-like CD4 T cell differentiation can be targeted and stimulated to generate Th1 cells in the presence of Tregs. Stem to Th1 CD4 differentiation is then sufficient to switch between active and restricted states and rescue response to immunotherapy.
