Fig. 6: Neoadjuvant cDC1-based vaccination enhances CD8⁺ and CD4⁺ memory T cell responses upon tumor recurrence.

a Experimental overview for (a–h): Mice grafted with MC38 cells (i.d.) were injected at day 3 and 8 with control PBS, anti-PD1 antibody or 10⁶ MC38-TCL loaded cDC1s (i.d), and tumors resected at day 10. 30 days later, mice were re-challenged with MC38 cells. Tumor naive mice only received MC38 cells at day 40. Secondary tumors were analyzed on day 5 after grafting. b Frequency of CD8⁺ or CD4⁺ T cells in secondary MC38 tumors (n = 6 or 7 [cDC1] biological replicates/group of one representative of two independent experiments). c CD45⁺ and CD8⁺ or CD4⁺ T cells sorted from secondary MC38 tumors and analyzed by scRNAseq (2 replicates per condition). Multiparametric t-SNE representation and annotation is shown. d Violin plots of tissue resident memory (Trm) hallmark gene expression within most abundant T cell clusters from (c). e t-SNE plot with distribution of T cells for each experimental group. Grey dots show all detected cells, black to yellow-colored areas show T cell density/experimental group. f Frequency of the most abundant clusters within secondary MC38 tumors (n = 2 biological replicates pooling 3 mice/group). g Experimental overview for (h, i): XCR1^DTRvenus mice received diphtheria toxin (DT) at days −2, −1, 0, 2, 5 i.p. and 10⁶ B16-OVA-TCL loaded cDC1s or PBS at days 0 and 5 (i.d. in ear) before analysis of ear skin at day 30. Frequency of CD4⁺ CD69⁺ CXCR6⁺ and CD4⁺ CD69⁺ CD103^- Trms (h, n = 4 [control], 5 [cDC1], 5 [cDC1+DT] biological replicates/group, representative experiment from 2 independent experiments) and representative flow cytometry plots (i, gated on CD45⁺ CD4⁺ cells) are shown. j Ranked pathway enrichment analysis comparing gene expression of CD4⁺ Trms with cells of CD4⁺ Tem and CD4⁺ IL7R clusters. k Violin plots showing expression of genes significantly (adj p value ≤0.001) enriched in CD4⁺ Trms of analysis in (j). Data are presented as mean ± SEM (dots represent individual data points) unless indicated otherwise. Statistical analysis by one-way ANOVA and Tukey post-hoc test (b, h) and Wilcox test (k). Source data are provided as Source Data file.