Fig. 6: (S)-CRZ and (R)-CRZ reduce lipid peroxides in CD8⁺ T cells to promote anti-tumor effect.

A (S)-CRZ (1 μM) or Lip-1 (1 μM) do not suppress the proliferation of B16-F10 cells, whereas (R)-CRZ (1 μM) exhibits cytotoxicity to B16-F10 cells. Statistical significance was assessed by comparing each group with the DMSO group at 96 h, distinguished by color. B Lip-1 (2 µM), (S)-CRZ (2 µM) and (R)-CRZ (2 µM) treatment reduce lipid-ROS in CD8⁺ T cells isolated from TdLNs and nTdLNs of B16-F10 tumor subcutaneous bearing mice. Lymph cells from lymph nodes separated from tumor-bearing mice were treated with indicated compounds for 3 h followed by flow cytometry analysis. Lipid-ROS was detected by fluorescence of C11-BODIPY 581/591 at the FITC channel. Data in A, B were presented as the mean ± S.D., n = 3 independent repeats. (C–F) Intraperitoneal injection of (S)-CRZ and (R)-CRZ suppress growth of B16-F10 xenograft tumors. B16-F10 tumor growth with indicated treatment (DMSO; 5 mg/kg (S)-CRZ; 5 mg/kg (R)-CRZ; 10 mg/kg Lip-1) (C). Body weight of mouse with indicated treatment (D). Photograph of B16-F10 tumors after excision from each group (E). Tumor weight of different treatment groups after excision (F). G (S)-CRZ (5 mg/kg) and (R)-CRZ (5 mg/kg) treatment significantly reduce lipid-ROS in CD8⁺ T cells from TdLNs of B16-F10 tumor subcutaneous bearing mice. CD8⁺ T cells were isolated from TdLNs of tumor-bearing mice with intraperitoneal drug treatment for 16 d. Lipid-ROS was detected by fluorescence of C11-BODIPY 581/591 in the FITC channel. Data in C–G are presented as the mean ± S.D., n = 5 independent repeats.