Fig 2: IL-2 Controls Cytotoxic CD4+ T Cell Differentiation in a Blimp-1-Dependent Manner(A) Prdm1fl/fl and Prdm1-/-Cd4cre mice bearing MCA205 tumors were treated or not with aCTLA-4 and monitored for tumor growth and survival.(B and C) TILs and dLNs were isolated at day 12 post-tumor inoculation for quantification of (B) GzmB-expressing cells within CD4eff and CD8 TILs and (C) T-bet-expressing cells within CD4eff and CD8 TILs (N = 9–11/group in two experiments).(D) Purified CD4+ T cells from dLNs and tumors from MCA205-bearing Prdm1fl/fl and Prdm1-/-Cd4cre mice were transferred to MCA205-bearing Rag1-/- mice at day 3 post-inoculation followed by aCTLA-4. Overall survival is shown (N = 5/group).(E) Quantification of CD25- and IL-2-stimulated pSTAT5-expressing CD4eff TILs (N = 5–11 group in two experiments).(F) Prdm1fl/fl and Prdm1-/-Cd4cre MCA205 tumor-bearing mice were treated with aCTLA-4 alone or in combination with high-dose intratumoral IL-2. TILs and dLNs were isolated at day 12 post-tumor inoculation for quantification of GzmB- and Prf-1-expressing cells within CD4+ TILs (N = 5/group).(G) Purified CD4+ T cells from WT (Prdm1fl/fl) and Prdm1-/-Cd4cre mice were transduced with Trp1 TCR-expressing vector and transferred to B16-bearing mice alone or in combination with aCTLA-4 + RT.(H) Representative plots and quantification of GzmB- and IFN-?-expressing cells within Trp1 effector TILs (N = 6/group in two independent experiments).(I) Transduced WT and Blimp-1-deficient Trp1 cells co-transferred 1:1 to the same host. Representative plots and quantification of GzmB-and Prf-1-expressing cells within Trp1 TILs are shown (N = 5/group).(J) Transduced cells as in (H) were transferred at day 8 to B16-bearing WT or Ifngr-/- mice alone or in combination with aCTLA-4 + RT. Cumulative tumor growth and survival are shown (N = 5/group).All quantification plots show mean ± SEM (one-way ANOVA) (J, two-way ANOVA).

Fig 3: Single-cell gene expression analysis confirms the importance of ADNP for IL-13 expression(A) UMAP plot of single-cell gene expression analysis of naive CD4+CD44-CD62L+CD25- T cells and CD4+CD44+CD62L-ST2+ effector T cells purified from the lungs of papain-challenged Cd4Cre and Cd4CreAdnpfl/fl mice.(B) UMAP plot with expression (log2 expression) of indicated genes per individual cell.(C and D) Volcano plot comparing genes from naive T cells (cluster 1) (C) and Th2 cells (cluster 2) (D) from Cd4Cre and Cd4CreAdnpfl/fl (KO) mice.(E) UMAP plot with expression (log2 expression) of Il13 per individual cell.(F) Violin plots with expression (log2 expression) of indicated genes. Rectangle, solid line, and dashed line represent the interquartile range, the median and the mean, respectively.(G) Heatmap showing the expression of indicated genes in cells from cluster 2 from Cd4Cre (left) and Cd4CreAdnpfl/fl (right) mice. Columns represent cluster 2 individual cells and rows represent the different genes.(H) UMAP plot showing cluster 2. Cells are colored by their cytokine expression pattern as shown, with number of cells indicated in parentheses.See also Figure S3.