Fig. 6

The rWTC-MBTA vaccine stimulates anti-metastasis adaptive immunity. Spleens were collected and isolated at the endpoint of the 4T1 subcutaneous metastasis animal model (a). CD4⁺ and CD8⁺ T cells were examined by flow cytometry in splenocytes from different treatment groups. b, Percentage of T-cells (CD4 ⁺ or CD8⁺) in splenocytes from different groups. c, Percentage of central memory (CD44⁺CD62L⁺) T cells in splenocytes of different treatment groups. d, Percentage of effector memory (CD44⁺CD62L⁻) CD4 and CD8 T cells in splenocytes. e-i, All the analyses are based on the co-culture 4T1 tumor cells and indicated splenocytes from different treated animals. e, 4T1 tumor cell number count after co-culture. f & g, Concentrations of cytokines IFN-𝛾 (f) and TNF-𝛼 (g) from co-culture supernatants as measured by ELISA. h&i, Percentage of Gran B⁺, IFN-𝛾⁺, TNF-𝛼⁺, or CD107⁺ CD4⁺ (h) or CD8⁺ T cells (i) was determined by flow cytometry. The analysis from e to i was conducted after co-culturing splenocytes (from different treated mice) with 4T1 tumor cells. Ordinary one-way ANOVA with multiple was used to assess statistical significance. All data are represented as mean ± SD. P-values are shown for each treated group compared to the rWTC-MBTA vaccine group (n≥3/group). *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001