【摘要】 目的探讨香加皮羽扇豆烷乙酸酯(CPLA)对人外周血单个核细胞(PBMC)来源的树突状细胞(DC)在体外分化成熟及免疫活性的影响。方法从人外周血分离单个核细胞,与细胞因子GM-CSF、IL-4共培养,于第5天加入DC的促成熟刺激剂TNF-α(阳性对照组)或CPLA。倒置显微镜和透射电镜下观察DC的形态;应用流式细胞术检测成熟DC的表面标志CD1a、CD83、CD80和CD86的表达情况;用ELISA检测DC培养上清中IL-12和IFN-γ的含量;用MTT法测定DC刺激T细胞增殖的能力。结果培养10d后,经CPLA刺激的PBMC呈现出典型DC的形态学特征;成熟DC的特征性表面分子CD1a、CD83、CD80和CD86表达水平均明显上调(P<0.05);细胞培养上清中IL-12和IFN-γ含量明显增高(P<0.05);刺激T细胞增殖的能力明显增强(P<0.05)。结论CPLA可诱导PBMC来源的DC分化成熟,并可促进其细胞因子的分泌,增强DC的免疫调节活性。更多还原

【Abstract】 AIM: To investigate the effect of lupane acetate of cortex periplocae (CPLA) on the differentiation, maturation and immune activity of human peripheral blood mononuclear cell (PBMC)-derived dendritic cells (DCs) in vitro. METHODS: PBMC isolated from human peripheral blood was cultured with GM-CSF, IL-4 for 5 d and stimulated with TNF-α (as positive control) or CPLA to induce DCs. The morphological characteristics of DC were observed under inverted microscope and transmisson electron microscope. The expressions of CD1a, CD83, CD80 and CD86 were analyzed by flow cytometry. IL-12, IFN-γ production in the culture supernatant of DCs was detected by ELISA. MTT method was used to determine the proliferation of T cells stimulated by DCs. RESULTS: After 10-days culture with cytokines and CPLA, PBMC developed into mature DCs with typical morphological characteristics and high expressions of CD1a, CD83, CD80 and CD86 on the cellular surface (P<0.05). CPLA enhanced IL-12 and IFN-γ production by DCs (P<0.05). CPLA-treated DCs markedly stimulated proliferation of T cells (P<0.05). CONCLUSION: CPLA may induce the differentiation and maturation of DC, up-regulate cytokines production and increase the immune activity of DC.[更多还原