人外周血树突状细胞诱导Th1/Th2细胞分化的研究

Research on human peripheral blood dendritic cell Inducing Th1/Th2 differentiation

目的 :研究人外周血树突状细胞 (DC)与Th0细胞在不同比例条件下 ,诱导Th0向Th1/Th2分化的关系。方法 :健康人外周血单个核细胞经传统的方法诱导得到DC ,将DC与自体的Th0细胞以 1∶4和 1∶30 0的比例混合培养 4天 ,收集Th细胞 ,经CD3和CD2 8抗体扩增 4 8h ,ELISA法测上清中IFN γ和IL 4的浓度。 结果 :从外周血中诱导出DC高表达CD83(73.4 % )、CD86 (90 .3% )、HLA DR(6 8.5 % )和CD4 0 (73.7% ) ,DC与Th0以 1∶4混合 ,产生的IFN γ与 1∶30 0混合组比有显著差异〔(34± 4 .3) μg/Lvs(3.5± 1.2 ) μg/L〕 ,产生的IL 4亦有显著差异〔(2 5 ± 4 .3)ng/Lvs (35 0± 12 0 )ng/L〕。 结论 :人外周血DC与Th0高比例混合时 ,诱导Th0向Th1分化 ;低比例混合时 ,诱导Th0向Th2分化。显示人外周血DC在调控T淋巴细胞免疫应答类型上具有可塑性 ,为临床移植物抗宿主病 (GVHD)免疫干预治疗奠定基础。

Objective: To study human peripheral blood dendritic cells(DC) inducing Th0 differentiation to Th1/Th2 at different DC/Th0 ration。 Methods: Mononuclear cells in healthy human peripheral blood were induced to DC through tradition method,4 days of culture of mixed DC/Th0 different ration( 1∶4 and 1∶300),harvesting Th,after 48 h of CD3 mAb and CD28 mAb expantion,in supernant IL-4 and IFN-γ measurement were determined using ELASA。 Results:Expresse CD83(73.4%), CD86 (90.3%), HLA-DR(68.5%),CD40(73.7%).DC/Th0 ration(1∶4) produced IFN-γ (34±4.3) μg/L, a little amount of IL- 4(25±4.3) ng/L. DC/Th0 ration(1∶300) produced IFN-γ(3.5±1.2) μg/L, a large amount of IL- 4(350±120) ng/L. Conclusion:Highly mixed DC/Th0 ration induced Th0 differentiation to Th1, low mixed DC/Th0 ration induced Th0 differentiation to Th2, DC regulating immunology response type were plastic which provided the basis for clinic GVHD therapy.