Additional file 1 of Peptide vaccine-conjugated mesoporous carriers synergize with immunogenic cell death and PD-L1 blockade for amplified immunotherapy of metastatic spinal

Abstract

Additional file 1: Fig. S1. Scanning electron microscopy image of as-prepared UCMS. Fig. S2. Purity of IDO peptide AL-9 determined by HPLC. Fig. S3. Mass spectrometric analysis of IDO peptide AL-9. Fig. S4. Fourier transform infrared spectra of as-prepared UCMS, UCMS@Pep, and UCMS@Pep-aPDL1. Fig. S5. (A) The fluorescent emission and (B) intensity value of FITC-labeled aPDL1 solution at different concentrations. Fig. S6. CLSM images of tumor cells treated with the complete culture medium and UCMS@Pep-RB for 4 h. The green fluorescence was emitted by UCMS@Pep-RB under irradiation with a 980 nm laser. Fig. S7. The cumulative release of RB from UCMS@Pep-aPDL1 under pH 7.4 and 5.5 at different time points. Fig. S8. Mean fluorescence intensity (MFI) of CRT expression in vitro (ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, n=3) Fig. S9. MFI of HMGB1 expression in vitro (ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, n=3)). Fig. S10. The blood circulation of UCMS@Pep-aPDL1 in mice after intravenous injection as determined by measuring Fe element at different time intervals (n=3). Fig. S11. Accumulation of UCMS@Pep-aPDL1 labeled by Cy7 in major organs and tumors and the related MFIs (n ≥ 3). Fig. S12. Results of routine blood count in mice treated with PBS and UCMS@Pep-aPDL1 (n ≥ 3). Fig. S13. Weight of tumors in every group. (*P < 0.05, **P < 0.01, ***P < 0.001, n≥3). Fig. S14. Overview of gating strategy for DCs (A) and Gating strategy of DCs (A) and T cells including CD4+ and CD8+ T cells and regular T cells (B). For DCs, briefly, firstly gating FSC-A versus FSC-H for single cells, followed by Live cells based on Zombie UV-negative cells (A, a-b). From viable cells, DCs (CD45+CD11c+) can be determined (A, c-e). Within the CD11c+ DCs, the maturation of DCs (CD80+CD86+) can be determined. For T cells, briefly, viable cells were determined by firstly gating FSC-A versus FSC-H for single cells, followed by lymphocytes based on FSC-A and SSC-A, and lastly CD45+ and Zombie UV-negative cells (B, a-c). From lymphocytes, CD4+ T cells (CD3+CD4+CD8−), CD8+ T cells (CD3+CD4-CD8+) can be determined (B, d-f). Within the CD4+ T cells, regulatory T cells can be determined as CD4+CD25+. Fig. S15. The quantitative results of DCs (A), CD4+ and CD8+ T cells (B,C) and Treg (D) filtration in tumor tissue (*P < 0.05, **P < 0.01, ***P < 0.001, n≥3). Fig. S16. Flow cytometric analyses of DCs, CD4 and CD8 T cells and Treg cells in the spleens of mice immunized using different interventions. Fig. S17. Kyn/Trp ratio in blood serum and tumor tissue. (*P < 0.05, **P < 0.01, ***P < 0.001, n≥3).