Extended Data Fig. 10: IL-10 expression promotes stemness in mouse and human CAR-T cells.

a, The experimental timeline for Fig. 6a-c. b-d, The experimental setting is described in Fig. 6d (n = 5 mice). Shown are frequencies of CD62L^hiCD44^lo cells among total CAR-T cells (b) and Sca-1⁺CD122⁺ cells among CD62L^hiCD44^lo CAR-T cells (c) in blood. d, Shown are the frequencies of IL-7Rα⁺KLRG-1^– among total CAR-T cells in blood. e-i, BALB/c mice were inoculated with 4T1-EGFRvIII-Luc (5 × 10⁴, i.v.), sublethally lymphodepleted by irradiation on day 5, and received i.v. adoptive transfer of IL-10 EGFRvIII CAR-T cells (3 × 10⁶), or EGFRvIII CAR-T cells (3 × 10⁶) on day 6 (n = 5 mice). On day 18, mice were killed for phenotype analyses of CAR-T cells in spleen and bone marrow by flow cytometry. e, Experimental timeline. f, h, Average frequencies of CD62L^hiCD44^lo cells among total CAR-T cells in spleen (f) and bone marrow (h). g, i, Average frequencies of Sca-1⁺CD122⁺ cells among CD62L^hiCD44^lo CAR-T cells in spleen (g) and bone marrow (i). j, The experimental setting is described in Fig. 6d. Shown are single-cell expression of key marker genes over the UMAP representation of the map. k-m, NSG mice were inoculated (s.c.) with PANC1-CD19 cells (2 × 10⁶) and received i.v. adoptive cell transfers of CD19 hCAR-T cells (1 × 10⁶) or IL-10 CD19 hCAR-T cells (1 × 10⁶) on day 8 (n = 4 mice). On day 63, mice were killed for phenotype analyses of CAR-T cells in spleen and bone marrow by flow cytometry. k, Experimental timeline. l, m, Shown are counts of CD45RA⁺CD27⁺CD62L⁺CD95⁺CCR7⁺ human CD8⁺ CAR-T cells in spleen (l) and bone marrow (m). n, The experimental setting is described in Fig. 6k. Shown are single-cell expression of key marker genes over the UMAP representation of the map. All data represent the mean ± s.e.m. and are analyzed by two-tailed Student’s t-test (f, g, h, i, l, m), one-way ANOVA with Tukey’s multiple-comparisons test (b-d).

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