Consistent with H&E staining results, TUNEL immunofluorescent staining also Comparing with other treatments, the expressions of T cells cytokines IL-2 increased mostly in the EH@Ag2S +US-treated group. In order to further verify the immunomodulatory effect in different groups, IFN-γ and CRT were also detected. It is well-known that the activated CD8 + T cells can secrete immune cytokines like interferon-γ (IFN-γ) to kill tumor cells. In addition, calreticulin (CRT) is an important biomarker of ICD. The IHC staining results showed that EH@Ag2S +US-treated group led to an increase in INF-γ and CRT expression, suggesting that they had the potential to induce an immune response through EH@Ag2S-mediated both the immune checkpoint blockade therapy and sonodynamic therapy. These results suggested that the synergistic therapy showed more effective cytotoxic T cell response.