Figure 7.

Jianpi Huayu Decoction (JHD) weakens the immunosuppressive ability of Myeloid-derived suppressor cells (MDSCs) in vitro. (A) Relative expression of ROS in MDSCs was analyzed by flow cytometry (n = 3). Representative flow cytometry data and statistical diagram are shown. (B) JHD ameliorated the proliferation inhibitive ability of MDSCs on CD4⁺ T cells. Briefly, MDSCs which isolated from H₂₂ hepatocellular carcinoma bearing mice by MACS technique were pretreated with JHD (500μg/ml) or not. Naïve CD4⁺ T cells were isolated from 6-weeks BALB/c mice by Naïve CD4⁺T cells Isolation Kit, and activated by plate-coated anti‐CD3 antibody (5μg/ml, Biogems) and soluble anti‐CD28 antibody (2μg/ml, Biogems) for 12 hours. Then CD4⁺ T cells and MDSCs were co-cultured at different ratios (CD4⁺ T cells: MDSCs = 1:1, 1:2) for 48h, and CD4⁺ T cells co-cultured with DC (6 × 10^{^5} cells/well) as positive control. Proliferation of CD4⁺ T cell was measured by carboxyl fluorescein diacetate succinimide (CFSE) using flow cytometry, with the gate of CD4⁺T cells (n = 3). (C, D) Expression of STAT-3 and iNOS in spleen were measured by immunohistochemistry staining (×400 magnification, n = 5). Scale bar = 50μm. *: p< 0.05; **: p < 0.01; ns, not significant.