Fig 2: Involvement of ferroptosis in the Mn-MOF-induced improved tumor immunity. (A-I) The numbers of CD4+ T cells (A), CD8+ T cells (B), CD8+CD69+ T cells (C), CD8+IFN-γ+ T cells (D), CD8+GzmB+ T cells (E), CD11c+CD80+ dendritic cells (F), CD11c+CD86+ dendritic cells (G), MDSCs (H) and Tregs (I) in tumor tissues of H22 tumor-bearing mice after intravenous injection of PBS, Mn-TCPP, MOF or Mn-MOF at Zr dosage of 5 mg/kg (the corresponding Mn dosage was 1 mg/kg) in the presence or absence of intraperitoneal injection of UAMC-3203 at 20 μg/kg dosage, followed by US irradiation (1.0 MHz, 1 W/cm2, 50% duty cycle) at tumors for 10 min. The data are presented as mean ± s.d. (n= 5). *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 3: Apoptosis and improved tumor immune microenvironment induced by combination of anti-PD-1 antibody and Met@Man-MPs in organotypic slices from one liver cancer patient-derived tumor.a The ratio of apoptotic tumor cells (CD45− cells) after the tumor slices from patient-derived tumor were treated with anti-human PD-1 antibody, Met@Man-MPs derived from THP-1-origniated macrophages or Met@Man-MPs plus anti-human PD-1 antibody at the concentration of anti-PD-1 antibody and Met of 20 and 40 μg mL−1 in the presence of PBMCs for 36 h, respectively. Data are presented as mean ± s.d. (n = 3 biologically independent samples; one-way ANOVA followed by Tukey’s HSD post-hoc test). (b–g) Percentages of M1-like TAMs (b), M2-like TAMs (c), CD8+ T (d), CD8+CD69+ T (e), CD8+IFN-γ+ T (f), and CD8+GzmB+ T cells (g) in the tumor slices from patient-derived tumor after treatment with anti-human PD-1 antibody, Met@Man-MPs derived from THP-1-origniated macrophages or Met@Man-MPs plus anti-human PD-1 antibody at the concentration of anti-PD-1 antibody and Met of 20 and 40 μg mL−1 in the presence of PBMCs for 36 h, respectively. Data are presented as mean ± s.d. (n = 3 biologically independent samples; one-way ANOVA followed by Tukey’s HSD post-hoc test). Source data are provided as a Source Data file.

Fig 4: CD274-L2A-derived soluble PD-L1 is not immunosuppressive.(A) Quantification of soluble PD-L1 by ELISA in supernatants of B-3T3 and HEK293T cells retrovirally transduced with CD274-L2A. Mean (± SEM) concentration from three independent experiments are shown. (B) Coomassie Brilliant Blue stain, under native or reducing (β-ME/SDS) PAGE conditions, of serum-free supernatants from HEK293T, CHO and B3 cells transfected or not with CD274-L2A. (C–D) Primary CD8+ T cells were labelled with CTV and stimulated with CD3- and CD28-coated beads for 72 hr, alone or co-cultured with HEK293T, HEK293T.CD274v1 or HEK293T.CD274-L2A cells transfected with the indicated amount of plasmid DNA. T cells were stained for intracellular GzmB at the end of the culture period. Representative histograms and scatter plots are shown in C; quantification of CTVlo and GzmB+ cells of three healthy donors according to the amount of transfected plasmid DNA is shown in D. (E) Percentage of activated (CD25+CD69+) Jurkat cells in the presence of conditioned media from parental HBL-1 cells, IFN-γ-stimulated HBL-1 cells, parental B-3T3 cells or B-3T3.CD274-L2A transduced cells. Cells were stimulated with CD3- and CD28-coated beads for 24 hr, with 10 µg/mL of a PD-L1-blocking antibody added where indicated. Mean (± SEM) proportion from three independent experiments are shown. (F) Percentage of activated (CD25+CD69+) Jurkat.PDCD1 cells after co-cultured with parental HEK293T, HEK293T.CD274v1, or HEK293T.CD274-L2A cells. Cells were stimulated with CD3- and CD28-coated beads for 24 hr, with 10 µg/mL of a PD-L1-blocking antibody added where indicated. Mean (± SEM) proportion from three independent experiments are shown.

Fig 5: T/NK cell clusters in NPC.a t-SNE plot showing 10 clusters of 17,263 T/NK cells (indicated by colors). b t-SNE plot, color coding for the expression of the marker genes (gray to red) for the indicated cell subtypes. c Average expression of selected T cell function-associated genes of naïve markers, inhibitory receptors, cytokines and effector molecules, co-stimulatory molecules, and Treg markers in each cell cluster. d Potential developmental trajectory of CD4+ T cells (n = 5694) inferred by analysis with Monocle 2. Arrows show the increasing directions of certain CD4+ T cell properties annotated with the signatures shown in e. e Traceplots of (left) CD4+ T cell activation signature along activation component and (right) terminal differentiation signature along terminal differentiation component for the CD4+ T cells. Cells are projected along the component, with the blue line indicating the moving average of the expression of signatures (a sliding window of length equal to 5% of the total number of CD4+ T cells was used), and the shaded area displaying SEM. Signatures used are presented in Supplementary information, Table S9. f Potential developmental trajectory of CD8+ T cells (n = 6975) inferred by analysis with Monocle 2. Arrows show the increasing directions of certain CD8+ T cell properties annotated with the signatures shown in g. g Traceplots (as in e) of (left) CD8+ T cell activation signature along activation component and (right) terminal differentiation signature along terminal differentiation component for the CD8+ T cells. Signatures used are presented in Supplementary information, Table S9. h Spearman correlation between the activity of CD8+ T cells (n = 6975), as measured by average granzyme expression (GZMA, GZMB and GZMH), and the expression of CD8+ T cell-specific genes. Genes encoding known immune checkpoint molecules are highlighted in blue. CD4+ Tconv, conventional CD4+ T cell; CD8+ Tdys, dysfunctional CD8+ T cell; NK, natural killer. i Heatmap showing the activity of TFs in each T/NK cell subtype. The TF activity is scored using AUCell. j Peripheral CD8+ T cells and NK cells were transfected with negative control (NC), EOMES-specific, RUNX3-specific, or XBP1-specific siRNAs, followed by immunoblot analysis to determine protein expression of Eomes, Runx3, or XBP1. β-actin is the loading control. The experiments were repeated independently for three times with similar results. k Left, representative histograms depicting the expression of GZMB and perforin on peripheral CD8+ T cells transfected with NC and EOMES-specific (n = 6 donors, n = 5 independent experiments), RUNX3-specific (n = 6 donors, n = 6 independent experiments), and XBP1-specific siRNAs (n = 6 donors, n = 5 independent experiments). Right, percentage of GZMB+ or perforin+ cells in siNC and siEOMES (n = 6), siRUNX3 (n = 6), and siXBP1 (n = 6) CD8+ T cells. l Left, representative histograms depicting the expression of GZMB and perforin on peripheral NK cells transfected with NC and EOMES-specific (n = 6 donors, n = 5 independent experiments), RUNX3-specific (n = 6 donors, n = 5 independent experiments), and XBP1-specific siRNAs (n = 6 donors, n = 5 independent experiments). Right, percentage of GZMB+ or perforin+ cells in siNC and siEOMES (n = 6), siRUNX3 (n = 6), and siXBP1 (n = 6) NK cells. *P < 0.05, **P < 0.01 (paired Student’s t-test).