Figure 6. The concentrations of cytokine profiles in ascites and kinetic changes of various lymphocytes in splenocytes of WF-3-challenged mice treated with PBS or CD25 monoclonal antibody.

(A) Concentrations of various cytokines in ascites. A1, IL-6; A2, IL-12; A3, TNF-α; A4, IFN-γ; A5, TGF-β. Note: Compared to mice treated with PBS only, the concentrations of cytokines like IL-6, IL-12, TNF-α, and IFN-γ were significantly elevated in mice treated with kinetic low-dose CD25 antibody than those in the other groups. The concentrations of TGF-β significantly decreased in the kinetic low-dose CD25 Ab group compared to the other groups. (B) Percentages of various lymphocytes. B1, CD3⁺CD4⁺ help T lymphocytes. Note: The percentages of CD4⁺ T lymphocytes in the splenocytes were not different between the PBS and CD25 Ab depletion groups on day 14 after WF-3 tumor challenge. However, the kinetic low-dose CD25 Ab depletion group had higher percentages of CD4⁺ helper T cells than the PBS and the other two sequential CD25 Ab depletion groups after 49 days of tumor challenge. B2, CD3⁺CD8⁺ cytotoxic T lymphocytes. Note: The percentages of CD8⁺ T cells were not different between the PBS and CD25 Ab depletion groups on 14 days after tumor challenge. The kinetic low-dose CD25 Ab depletion group had higher percentages of CD8⁺ cytotoxic T cells than the other groups on day 49 after tumor challenge. B3, CD4⁺CD25⁺Foxp3⁺ Treg cells. Note: The percentages of Treg cells in the splenocytes were not different among these groups on day 14 after tumor challenge. However, the kinetic low-dose CD25 Ab depletion group had lower percentages of Treg cells than the other groups on day 49. B4, ratios of cytotoxic T lymphocytes/Treg cells. Note: The ratios of CD8⁺ cytotoxic T cells/Treg cells in the kinetic low-dose CD25 Ab depletion group were highest among the four groups. (C) Mesothelin-specific IFN-γ ELISPOT assays of splenocytes in the PBS and CD25 Ab-treated groups. C1, the representative figures of mesothelin-specific IFN-γ ELISPOT assays of splenocytes; C2, bar figures of the numbers of IFN-γ-secreting, mesothelin-specific T lymphocytes in various groups. Note: The kinetic low-dose CD25 Ab depletion group generated the highest numbers of IFN-γ-secreting, mesothelin-specific T lymphocytes compared to the other groups. (D) Survivals of mice treated with PBS, neutralizing TGF-β, neutralizing IL-10 or CD25 monoclonal antibodies. D1, Diagrammatic representation of the different treatment protocols of neutralizing TGF-β, neutralizing IL-10 or CD25 monoclonal antibody; D2, Survival curves of mice treated with PBS, neutralizing TGF-β, neutralizing IL-10, neutralizing TGF-β and neutralizing IL-10 or CD25 monoclonal antibody. Note: The mice treated with kinetic low-dose CD25 Ab had the longest survival time after WF-3 tumor challenge when compared to the other mice treated with PBS, neutralizing TGF-β, neutralizing IL-10 or neutralizing TGF-β and neutralizing IL-10 monoclonal antibodies.