消瘰丸治疗淋巴瘤基于PD-1/PD-L1信号通路的机制研究

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。

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

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.