Extended Data Fig. 4: NRP1 modulates the memory differentiation of tumor-primed effector CD8⁺ T cells but not effector polyfunctionality.

a–d, CD8⁺ T cells from primary B16-gp100 tumors and matched spleens were analyzed on D12 (n = 8 per genotype). (a) Numeration of total D^b-gp100⁺ cells, as well as their subsets (T_CM, T_EFF, MPECs, SLECs) within intratumoral CD8⁺ T cells; (b) Ratio between intratumoral Db-gp100⁺CD8⁺ MPECs and SLECs; (c-d) Representative flow cytometry plots (c, left) and quantification for the frequencies of D^b-gp100⁺ CD8⁺ T cells (c, right) and (d) Db-gp100⁺ T_CM in CD8⁺ T cells from spleen; e, The incidence (left) and growth (right) of 2° B16-gp100 tumor (+30d rechallenge). f, Representative flow plots for the expression of IRs on CD8⁺ TILs of 1° B16 tumors on D12 (left) and SPICE plots (right) visualization for co-expression of multiple IRs. g, Numeration of 5-IR-co-expressing cells (5-IR⁺) on D12 (n = 5), by frequency within CD8⁺ TILs (left) and absolute number per gram tumor mass (right). Data were representative from 2 independent cohorts. h, i Expression of cytokines (IFN-γ against GzmB, TNF-α and IL2, respectively) by the CD8⁺ TILs on D18 of 1° tumor. SPICE plots visualization for the multi-cytokine producing cells. i, Percentage of single, dual, or multi-cytokine producing cells within the CD8⁺ TILs as shown in h, n = 6 per genotype. Error bars, mean±s.e.m; Statistical significance was determined by two-tailed unpaired Student’s t test (a–d, g,i) or log-rank test (e, left) or two-way ANOVA with correction for multiple comparisons (e, right).

Source data