To investigate alterations in the tumor microenvironment after TRT and immunotherapy and thus further elucidate the potential mechanisms of TRT combined with immunotherapy, PD-L1+ immune cells (CD45+/PD-L1, CD11b+/PD-L1), PD-L1+ neoplastic cells (CD45-/PD-L1), CD8+ T cells and tumor-infiltrating regulatory T cells (CD4+/Foxp3+) were assessed dynamically using both flow cytometry and multiplexed immunofluorescence. Here, FOXP3 is linked to neoplasm.