目的研究人参皂苷Rg3联合细胞因子对单核细胞来源树突状细胞(DC)的诱导及其免疫功能的影响。方法选取8例健康志愿者分离外周血单核细胞,利用人参皂苷Rg3联合细胞因子(GM—CSF、IL-4、TNF-α)培养14d诱导Dc,光镜和电镜观察细胞形态,用流式细胞仪检测细胞免疫表型,同种异体混合淋巴细胞反应(MLR)检测其抗原呈递能力。结果(1)单核细胞经人参皂苷Rg3联合细胞因子诱导均产生了不同比率的具有树突状细胞形态学特征的DC。这些细胞表达DC的表面分化抗原,能刺激产生MLR反应.(2)Rg3(5.0μg/ml)联合细胞因子(GM.CSF150ng/ml,IL-450ng/ml,TNF—α 40ng/mt)使单核细胞-DC获得率(以CD80、CD86双阳性定义DC)由43.31%提高到59.75%,并提高了DC刺激淋巴细胞的增殖能力。结论人参皂苷Rg3联合细胞因子可以将正常人单核细胞体外诱导成为形态、表型和功能符合DC特征的细胞,可使单核细胞诱导DC产率增加,并提高其抗原呈递功能。
Objective To investigate the impact of Rg3 combined cytokines in inducing dendritic cells (DCs) generation from monocyte and enhancing antigen presentation. Methods Isolated peripheral blood mononuclear cells from 8 healthy volunteers, using the Rg3 combined cytokines ( GM-CSF, IL-4, TNF-αct) cultured for 14 days to induce DC. Optical and electron microscopy were used to observe the morphologie features of DCs. Immunophenotype of DCs were analyzed by flow cytometry. The function of antigen presentation was observed by allo- mixed lymphocyte reaction (MLR). Results ( 1 ) Rg3 and cytokins in all groups could induce DCs with typical dendritic morphology. The generated cells expressed the surface differentiation antigens of DC, had the function of DC, and could stimulate MLR. (2) Rg3 in combination of cytokins had the ability to enhance DCs generating from monocytes,the generating rate increased from 43.31% to 59. 75%. At the same t/me Rg3 in combination of cytokins could enhance lymphocyte proliferation stimulated by DCs. Conclusions Rg3 in combination with cytokins could induce monocytes from health adult to DC. Ginseng saponin Rg3 can increase DCs production and enhance their antigen presentation.