Extended Data Fig. 4: TCF1+lin- CD4 T cells are stem-like cells that are actively restrained in the tumor response.
From: Differentiation fate of a stem-like CD4 T cell controls immunity to cancer
a) Experimental design to characterize SMARTA differentiation kinetics. b-e) Total numbers and representative phenotype of recovered SMARTAs in TDLNs and tumor 1- to 5-weeks after transfer in TRAMPC1-GP mice. Summary graphs show median or mean ± s.d. Data are representative of 1-2 independent experiments for each timepoint (n = 4–6 mice per group for each timepoint). f) Experimental design and total number of recovered SMARTAs in TDLNs and tumor 4-weeks post transfer. g) Representative histogram of PD-1 expression on recovered SMARTAs in TDLNs and tumor as compared to naïve CD4 T cells. h-i) Phenotypic analysis of SMARTAs in TDLNs and tumor 4-weeks post transfer (n = 9). Medians are shown for each summary plot. j) Representative phenotype of endogenous GP66 + CD4 T cells (top) or transferred SMARTAs (bottom) in the indicated tissue 8 days after LCMV Armstrong infection. Summary plots show median of the total number of GP66+ or SMARTAs in each tissue. k) Representative histograms for Th1 (TBET + TCF1-) and Tfh (TBET- TCF1 + CXCR5 +) populations for the endogenous GP66+ or SMARTAs in the spleen 8 days after LCMV Armstrong infection. Naïve (CD44- PD1-) CD4 T cells are plotted as a reference for each marker. Data are representative of 2 independent experiments (n = 8 recipient mice). l) Experimental design to test how PDL1 therapy affects stem-like CD4 differentiation in TRAMPC1-GP refractory tumor model. m) Tumor kinetics as shown by tumor diameter shown as mean ± s.d. for Untx and aPDL-1 treated mice and analyzed by two-sided unpaired Mann–Whitney U test (n = 5–7 mice per group). n) Phenotype of GP66 + CD4 T cells in TDLNs (top) and tumor (bottom) in Untx or aPDL-1 treated mice 14-days after treatment in TRAMPC1-GP bearing mice. Mean ± s.d. represented and analyzed by two-sided unpaired Mann–Whitney U test (n = 5–7 mice per group). o) Tumor kinetics shown as mean ± s.e.m. and phenotype of PD1 + CD4 T cells for Untx and PDL-1 treated mice in MC38 responsive tumor model. Median shown for phenotype summaries. Statistical comparisons were analyzed by two-sided unpaired Mann–Whitney U test (n = 7 mice per group). p) Phenotype of bulk activated CD4 T cells for untreated and PDL-1 treated mice in RENCA-HA responsive tumor model. Mean ± s.d. are represented and analyzed by two-sided unpaired Mann–Whitney U test (n = 4 mice per group).
