Fig 2: C-agarose+CpG Treatments Change the Ratio of Immune Cell Subsets and Cytokine Levels in the TDLN from a Murine 4T1 Breast Cancer Model(A) Lymph node cells were stained with PE-CD4 (CD4+ T cells), APC-CD8 (CD8+ T cells), APC-CD11c (DCs), or PE-CD49b (NK cells) and examined by flow cytometry. (B) The ratio of CD4+ T cells, CD8+ T cells, DCs, and NK cells. (C) The homogenates of lymph nodes were harvested to determine the levels of IL-1α, IL-1β, TNF-α, CXCL9, CXCL10, IL-2, IFN-γ, and granzyme B by ELISA. n = 10 mice per group. Data are representative results from five independent experiments and are expressed as the mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001. m-ODN, mismatched oligonucleotide; TDLN, tumor-draining lymph node.

Fig 3: SDT plus aPD-L1 therapy activating systematic antitumor immunity. A Representative flow-cytometry plots showing the tumor-infiltrating leucocyte cells, including CD45 + cells (CD45 +) (A), CD8 + T cells (CD45 + CD3 + CD8 +) (B) and NK cells (CD45 + CD3-CD49b +) (C) in orthotopic tumors and the quantitative data (D–F). The tumor sections were stained using H&E, TUNNEL, Ki-67, and CRT (G) and the quantitative data (H–J)

Fig 4: The BacMam-based IL-15:IL-15Rα-B16F10-OVA vaccine induced strong tumor antigen-specific T cell and NK cell responses in mice. (A–C) The mice were immunized with IL-15:IL-15Rα-B16F10-OVA (IL-15:IL-15Rα BV), control-B16F10-OVA (control BV), or no virus-infected B16F10-OVA (no virus), as shown in Figure 3A (n = 4~5) and challenged with live B16F10-OVA cells. (A) Splenocytes were collected at day 22 and stained to analyze for the percentage of effector memory T cells (TEM, CD44+CD62L–), central memory T cells (TCM, CD44+CD62+), OVA-specific CD8+ T cells (H-2Kb/OVA (SIINFEKL)-Tetramer+CD3+CD8+), and the NK subset (total NK cells; CD49b+, effector NK; CD49b+CD11b+). (B) PBMC were harvested on day 17 and 22 and stained to analyze OVA-specific CD8+ T cells (H-2Kb/OVA (SIINFEKL)-Tetramer+CD3+CD8+). (C) The TILs from the vaccinated mice were collected at day 22 and stained to analyze effector memory and central memory T cells and NK subsets. Note: * p < 0.05, ** p < 0.01, OVA+CD8; OVA-specific CD8+ T cells.

Fig 5: Role of innate lymphoid cytotoxic cells in toll-like receptor (TLR)9-mediated protection against plasmacytoma growth. (A) Flow cytometry analysis of CD49b+ cells in pooled spleen cells from 2 BALB/c mice with and without treatment with 2 mg anti-ASGM1 antibody. Experiment representative of two independent experiments. (B) Survival of groups of BALB/c mice (n = 7) either mocked-treated (open circles) or treated only with CpG-ODN (closed circles), or with CpG-ODN and anti-ASGM1 treatment (closed squares), was monitored daily after i.p. administration of TEPC.1033.C2 cells. Experiment representative of three independent experiments. *P = 0.0153 and **P = 0.0015 by Log-rank (Mantel–Cox) test.