消瘰丸治疗淋巴瘤基于PD-1/PD-L1信号通路的机制研究

Mechanism research on the treatment of lymphoma with Xiaoluo pill based on PD-1/PD-L1 signaling pathway

[目的]初步探究古方消瘰丸治疗淋巴瘤基于程序性死亡因子(PD-1)/程序性死亡因子配体1(PD-L)信号通路上的免疫调控机制。[方法]从健康人、淋巴瘤患者的外周血中分离外周血单个核细胞,体外培养刺激后,应用流式细胞仪、CFSE等技术,以PD-1/PD-L1信号通路为靶通路,通过是否阻断PD-1/PD-L1信号通路与消瘰丸药物干预,观察消瘰丸对T细胞不同亚细胞的活性作用。[结果]淋巴瘤患者T淋巴瘤细胞在消瘰丸干预下,CD4^+T细胞PD-1(+)和CD69表达降低,而CD8^+T细胞PD-1(+)和CD69表达增加;PD-L1阻断剂和消瘰丸联合使用对淋巴瘤T细胞中的CD4^+细胞CD69抑制作用比PD-L1阻断剂组和消瘰丸组都明显;与PD-L1阻断剂组相比,消瘰丸组CD8^+T细胞活性增加更明显,而同时加入两者,与消瘰丸组相比,增加并不明显。[结论]消瘰丸对大量异常增殖的CD4^+T细胞可以抑制PD-1(+)的高表达,消瘰丸与PD-L1阻断剂二者对于CD4^+PD-1(+)和CD69表达有协同降低的作用;消瘰丸对于有细胞毒作用的CD8^+T细胞活性有增强作用,可以促进PD-1(+)的表达。

[Objective] To investigate the immunoregulatory mechanism based on PD-1/PD-L1 signaling pathway in the treatment of Lymphoma with Xiaoluo pill. [Methods] Peripheral blood mononuclear cells（PBMC） were isolated from the peripheral blood of healthy people and lymphoma patients. After the cells were cultured in vitro, the activity of different subcellular cells of T cells was observed by whether to block the PD-1/PD-L1 signal pathway and drug intervention, which was detected by flow cytometry, CFSE and other technologies based on PD-1/PD-L1 signal target pathway. [Results] The expressions of PD-1（＋） and CD69 were decreased in CD4^＋ T cells and increased in CD8^＋ T cells in patients with T lymphoma. The inhibitory effect on CD69 in CD4^＋ cells in lymphoma T cells in combination therapy of PD-L1 blockade and Xiaoluo pill was significantly higher than that in PD-L1 blockade group and Xiaoluo pill group alone. Compared with the PD-L1 blockade group, the activity of CD8^＋ T cells increased significantly in Xiaoluo pill group. And compared with Xiaoluo pill group, the activity of CD8^＋ T cells in combination therapy of PD-L1 blockade and Xiaoluo pill was not significantly increased. [Conclusion] Xiaoluo pill could inhibit the high expression of PD-1（＋） on a large number of aberrant proliferative CD4^＋ T cells;both Xiaoluo pill and PD-L1 blockade have a synergistic effect on PD-1（＋） and CD69 expression in CD4^＋ T cells; Xiaoluo pill has a potent effect on CD8^＋ T cell activity and could promote PD-1（＋） expression.