Fig 2: D665-G3c treatment induced permanent allograft acceptance.(A) Treatment protocol of D665 and G3c in mouse heart transplantation. D665 (250 μg per mouse) and G3c (250 μg per mouse) were intraperitoneally injected on days −3 and 0 sequentially. Heart transplantation was performed on day 0. (B) The survival of cardiac allografts in each group (n = 9 to 12 for each group). The graft survival of each group was evaluated using Kaplan-Meier curves and log-rank tests. ****P < 0.0001. (C) Representative hematoxylin and eosin staining of cardiac grafts in each treatment group on POD7 (n = 6 for each group). Scale bars, 100 μm. (D) A representative FCM analysis of CD4+CD25+Foxp3+ Treg cells in the splenocytes of each treatment group on POD3 and POD7. (E) A representative FCM analysis of IL-10/IFN-γ–co-producing CD4+ Tr1 cells in the splenocytes (SPs) and GILs of each treatment group on POD7 (n = 4 for each group). (F) A representative FCM analysis of CD4+ T and CD8+ T cells in the splenocytes and GILs of each treatment group on POD7 (n = 4 for each group). (G) Representative CD8 (blue), 5-bromo-2′-deoxyuridine (BrdU) (red), and type IV collagen (yellow) triple immunohistochemistry staining of heart grafts in each group on POD7 (n = 4 for each group). Scale bars, 100 μm. (H) Survival of cardiac allografts in each group (n = 6 for each group). The graft survival of each group was evaluated using Kaplan-Meier curves and log-rank tests. ***P < 0.001.

Fig 3: Agpat4 silencing in CRC suppresses T cell-dependent tumor progression. a Eight-week-old male NCG mice were intraperitoneally inoculated with sh-Agpat4- or sh-NC-transfected MC-38 cells (4 × 106 cells in 100 μl of PBS per mouse). Twelve days later, the mice were sacrificed, and the tumor nodules were collected and weighed. (n = 6). b Agpat4-silenced MC-38 cells increased the percentage of CD8+ T cells in epididymal fats. Six-week-old mice were intraperitoneally engrafted with sh-Agpat4- or sh-NC-transfected MC-38 cells (4 × 106 cells in 100 μl of PBS per mouse) on day 0 and were sacrificed for the analysis of total T cells, CD4+ T cells and CD8+ T cells in the stromal vascular fraction (SVF) of epididymal fats on days 0, 3, and 5. T cells were marked by the phenotype CD45+CD3+CD11b−. CD4+ T cells were defined by the phenotype CD45+CD3+CD11b−CD8−CD4+. CD8+ T cells were defined by the phenotype CD45+CD3+CD11b−CD4−CD8+. Representative results are shown. c Percentage of IFN-γ+ cells in the CD8+ T cells described above in b. (n = 5). d Six-week-old male C57BL/6 mice were intraperitoneally inoculated with MC-38 cells (4 × 106 cells in 100 μl of PBS per mouse) on day 0 and then treated with anti-CD8 antibody or IgG as a control 3 times (days 2, 4, and 6) at a dose of 100 μg/day. Mice were sacrificed for tumor observation on day 14. (n = 5). The data in a–d represent the means ± s.e.ms. (***P < 0.005; Student’s t test)

Fig 4: Colec11–/– mice exhibit less immunosuppressive TME.Tumors excised from Colec11+/+ (WT) or Colec11–/– (KO) mice (d14) were used for analyzing TME. (A–C) Tumor infiltrates analyzed by flow cytometry. (A) CD45+ cells. (B) Subsets of tumor-infiltrating leukocytes analyzed by flow cytometry. Data were analyzed by unpaired t test (n = 18 mice per group, pooled from 4 experiments). Each dot represents an individual mouse. (C) A bar chat representing proportion of subsets in CD45+ cells shown in B. (D) Representative microscopy images of immunochemical staining for CD11b (green)/CD3 (red)/DAPI (blue) and F4/80 (green)/CD3 (red)/DAPI (blue). Scale bar: 50 μm. (E) Representative microscopy images of immunochemical staining for CD8 (red)/DAPI (blue) in tumor edge and core areas. Scale bar: 50 μm. (F). qPCR analysis in tumor tissues. Data were analyzed by unpaired t test (n = 8 mice per group). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Fig 5: Systemic antitumor immunity induced by aAGd-NW-sensitized radiation.a Schematic illustration of aAGd-NWs sensitized radiation-mediated in situ vaccine. Created with BioRender.com. b–d Tumor growth curves (b), isolated tumor photographs (c), and tumor weights (d) of BALB/c mice immunized with RT and aAGd-NWs+RT treated tumor cells, respectively (n = 8 mice). e Flow cytometry analysis of CD11c+ and H2Kb-SIINFEKL+ DCs in TDLNs after Vehicle, GGd-NCPs, aAGd-NWs, Vehicle+RT, GGd-NCPs+RT, aAGd-NWs+RT treatments (n = 3 mice). f Flow cytometry analysis of DCs maturation (CD80+ and CD86+ gated within CD11c+) in TDLNs (n = 8 mice). g Tumor growth curves of CT26 colorectal tumor-bearing mice (n = 8 mice). Black and red arrows indicated drug/radiation and anti-CD8a/anti-PD-L1 treatments, respectively. RT (5 Gy × 2) was performed on day 0 and 6, respectively. Anti-CD8a/anti-PD-L1 antibodies (10.0 mg kg−1 × 4 with fractions delivered 3 days apart) were administered via intraperitoneal injection 6 h after X-ray irradiation. h, i Percentages of CD4+ (h) and CD8+ (i) T cells infiltrating within tumor tissues as detected by flow cytometry (n = 4 mice for the combinational group and n = 6 mice for other groups). j Relative multiples of IFN-γ secretion in the tumor tissues as detected by ELISA kit (n = 4 mice for the combinational group and n = 6 mice for other groups). All data were shown as mean ± SD. Two-tailed Student’s t-test was used to calculate statistical differences between two groups, and one-way ANOVA analysis of variance was used for multiple groups. p values > 0.05 represented nonsignificance (N.S.) and p values < 0.05 represented statistically significant. Source data are provided as a Source Data file.