Fig. 4. The hydrogel microsphere vaccine amplified the recruitment, activation and migration of CD103⁺CD11b⁻ cDC1s.

a Schematic diagram of the in vivo experiments. b Representative images of H&E staining and CD45-labelling IHC staining of pancreatic cancer tissues after different combination therapy. Five biologically independent samples were performed to confirm data stability. c Analysis of the intratumour release of FLT3L and CD40L by the hydrogel microsphere vaccine or direct intratumour injection of dual cytokines. n = 3 biologically independent samples. d Representative images of flow cytometry analysis showing subpopulations of tumour-resident DCs across different groups. e Quantification of the proportion of tumour-resident CD24⁺CD64⁻ total DCs and CD103⁺CD11b⁻ cDC1s across different treatment groups (n = 5 for each group). f Quantification of the expression of CD86 and CD40 on the surface tumour-resident cDC1s across different groups (n = 5 for each group). g Quantification of the cellular density of tumour-resident CD24⁺CD64⁻ DCs and CD103⁺CD11b⁻ cDC1s across different treatment groups (n = 5 for each group). The hydrogel microsphere vaccine significantly amplified the cellular density of tumour-resident cDC1s and increased the cDC1-to-cDC2 ratio in pancreatic cancer. h Identification of the correlation between the cellular density of tumour-resident total DCs or cDC1s and the tumour weight across all tumour-bearing mice. i, j Representative images of flow cytometry analysis showing total CD11c⁺MHC II⁺ myeloid cells and subpopulations of DCs in the TdLNs and spleens across different groups. k, l Quantification of the density of CD11c⁺MHC II⁺ myeloid cells and CD103⁺CD11b⁻ cDC1s in TdLNs (k) and spleens (l) in mice bearing pancreatic cancer. The hydrogel microsphere vaccine significantly amplified the density of cDC1s and increased the cDC1-to-cDC2 ratio in the TdLNs and spleens in mice bearing pancreatic cancer. m Representative images showing IF-stained CD103, CD31, CD4 or CD8-positive cells in pancreatic tumour tissue (left panel), TdLNs and spleens in tumour-bearing mice treated with the hydrogel microsphere vaccine or Group1 control microspheres. n Quantification of the density of tumour-resident OVA_257-246-H-2Kb-specific cDC1s in hydrogel microsphere vaccine-treated mice and Group1-treated mice. e–g, k, l, n n = 5 biological independent samples for each group. Data are presented as mean values +/− SEM. Two-tailed unpaired t test. h n = 25 biological independent samples, data are tested using Pearson’s test with two-tailed P values. Source data are provided as a Source data file.