Figure 2. CD4 T cells govern fate decisions of tumor-infiltrating monocytes.

(A) Experimental approach to test the role of T cell subsets on monocyte fate.

(B) Proportion (left) and number (right) of Tomato⁺ intratumoral macrophages. Each dot is an independent mouse. Data are mean ± SEM; n = 3–5 mice per group; *p < 0.05; one-way ANOVA with Tukey’s posttest.

(C) Proportion of MHCII^hi or MHCII^lo Tomato⁺ macrophages. Each dot is an independent mouse. Data are mean ± SEM; n = 3–5 mice per group; ****p < 0.0001; one-way ANOVA with Tukey’s posttest.

(D) Experimental approach to test the impact of prolonged CD4 T cell depletion in tumor-bearing CCR2^CreER R26^TdTomato mice. See also Figure S3.

(E) Tumor weight from mice in (D); n = 3–6 mice per group. Data are mean ± SEM; **p < 0.005; Student’s t test for each time point.

(F) Proportion of Tomato⁺ macrophages from (D) that are MHCII^hi or MHCII^lo. Data are mean ± SEM; n = 3–6 mice per group; ***p < 0.001, ****p < 0.0001; Student’s t test for each time point.

(G) Representative histograms gated on Tomato⁺ macrophages isolated from PBS- or αCD4-treated mice.

(H) Proportion of Tomato⁺ macrophages expressing Arg1, CD206, and FRβ from PBS- or αCD4-treated mice. Data are mean ± SEM; n = 3–6 mice per group;*p < 0.05, **p < 0.005, ****p < 0.0001; Student’s t test for each time point.

(I) Representative PD-L1 staining (top) and MFI of Tomato⁺ macrophages from PBS- or αCD4-treated mice (bottom); n = 3–6 mice per group; *p < 0.05; Student’s t test.

(J) Representative plots and proportion of Tomato⁺ MHCII^lo macrophages that coexpress CD206 and FRβ from mice in (D). Data are mean ± SEM; n = 3–6 mice per group; ****p < 0.0001; Student’s t test for each time point.

(K) Representative histograms and proportion of Tomato⁻ macrophages that are Ki67⁺. Data are mean ± SEM; n = 3–4 mice per group; **p < 0.005; Student’s t test.

(L) Experimental approach to test the impact of T cell antigen specificity on monocyte differentiation. See also Figures S4A–S4F.

(M) Tumor weight from (L). Data are mean ± SEM; n = 4 mice per group; **p < 0.005; Student’s t test.

(N) Proportion of MHCII^hi or MHCII^lo macrophages from (L). Data are mean ± SEM; n = 4 mice per group; ****p < 0.0001; Student’s t test.

(O) Proportion of Arg1⁺ macrophages from (L). Data are mean ± SEM; n = 4 mice per group; ***p < 0.001; Student’s t test.

(P) Proportion of MHCII^loCD206⁺FRβ⁺ macrophages from (L). Data are mean ± SEM; n = 4 mice per group; ****p < 0.0001; Student’s t test.

(Q) Experimental approach to test if transferred tumor-specific (OTII) or non-specific (SM1) CD4 T cells modulate monocyte differentiation in KPC-OVA-bearing Rag^−/− mice. See also Figures S4G–S4I.

(R) Proportion of MHCII^hi or MHCII^lo macrophages from (Q). Data are mean ± SEM; n = 4 mice per group; ***p < 0.001; Student’s t test.

(S) Proportion of Arg1⁺ macrophages from (Q). Data are mean ± SEM; n = 4 mice per group; **p < 0.005; Student’s t test.

(T) Proportion of MHCII^loCD206⁺FRβ⁺ macrophages from (Q). Data are mean ± SEM; n = 4 mice per group; **p < 0.005; Student’s t test. See also Figure S5.