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目的探讨细胞因子在CD4~+CD25~+调节性T细胞免疫抑制机制中的作用,观察胃癌患者外周血中的CD4~+CD25~+调节性T细胞及其效应性T细胞CD4~+CD25~-T细胞产生具有不同生物活性的细胞因子的水平。方法采用免疫磁珠分选方法分离胃癌患者外周血中的CD4~+ CD25~+T细胞与CD4~+CD25~-T细胞后,将CD4~+CD25~+T细胞与效应性T细胞CD4~+CD25~-T细胞在体外分别单独培养及按不同比例共同培养后,再用酶联免疫吸附试验(ELISA)法检测单独及混合培养时细胞因子干扰素(IFN)-γ、白细胞介素(IL)-10及转化生长因子(TGF)-β分泌的水平。结果健康对照及胃癌患者CD4~+CD25~+T细胞分泌抑制性细胞因子IL-10、TGF-β均显著高于CD4~+CD25~-T细胞,而分泌细胞因子IFN-γ均显著低于CD~+CD25~-T细胞(P＜0．01)。CD4~+ CD25~+T细胞在体外可明显抑制CD4~+CD25~-T细胞产生IFN-γ的能力,且这种抑制能力呈效靶比关系(P＜0．01)；但CD4~+CD25~+T细胞与效应性T细胞CD4~+CD25~-T细胞在体外共培养对IL- 10及TGF-β的分泌无显著影响(P＞0．01)。结论CD4~+CD25~+调节性T在体外可能通过细胞因子的调节发挥对效应性T细胞的抑制作用。
Objective To investigate the role of cytokines in the immunosuppressive mechanism of CD4 ~ + CD25 ~ + regulatory T cells and to observe the relationship between CD4 ~ + CD25 ~ + regulatory T cells and CD4 ~ + CD25 ~ + regulatory T cells in peripheral blood of gastric cancer patients. -T cells produce levels of cytokines with different biological activities. Methods CD4 ~ + CD25 ~ + T cells and CD4 ~ + CD25 ~ -T cells were isolated from peripheral blood of patients with gastric cancer by immunomagnetic bead sorting. CD4 ~ + CD25 ~ + T cells and CD4 ~ + CD25 ~ -T cells were cultured in vitro and co-cultured in different proportions, and then detected by enzyme-linked immunosorbent assay (ELISA) cytokine interferon (IFN) -γ, interleukin (IL- IL) -10 and transforming growth factor (TGF) -β secretion. Results The secreting cytokines IL-10 and TGF-β secreted by CD4 ~ + CD25 ~ + T cells in healthy controls and gastric cancer patients were significantly higher than those in CD4 ~ + CD25 ~ -T cells, while the secretion of cytokines IFN- CD ~ + CD25 ~ -T cells (P <0.01). CD4 ~ + CD25 ~ + T cells can significantly inhibit the production of IFN-γ in CD4 ~ + CD25 ~ -T cells in vitro, and this inhibitory capacity is in a dose-versus-effect relationship (P <0.01) Coculture of CD25 + T cells and effector T cells CD4 ~ + CD25 ~ -T cells had no significant effect on the secretion of IL-10 and TGF-β (P> 0.01). Conclusion CD4 ~ + CD25 ~ + regulatory T may exert an inhibitory effect on effector T cells through the regulation of cytokines in vitro.