目的观察不同细胞因子诱导对于体外培养的树突状细胞(DC)免疫刺激活性的影响。方法通过1000U/ml人粒细胞-巨噬细胞集落刺激因子(rhGM—CSF)和500U/ml白细胞介素(IL)4体外诱导的人单个核细胞来源的DC,经1000U/ml肿瘤坏死因子(TNF)-α、1mg/L脂多糖(LPS)、1mmol/L丁酸钠、细胞因子鸡尾酒法诱导成熟,分别以流式细胞仪、FITC—Dxtran内吞检测、混合淋巴细胞反应(MLR)、酶联免疫吸附试验(ELISA)检测DC的表面标志、内吞能力、刺激淋巴细胞增殖能力和IL-12分泌的变化。结果鸡尾酒法诱导下的DC成熟标志显著上调,内吞能力减弱186.74±38.66,刺激淋巴细胞增殖能力增强18.23±2.22,并且IL-12的分泌能力增加(656.18±38.52)ng/L,说明其显著促进DC成熟。而丁酸钠诱导下的各项成熟指标均下调,导致DC免疫刺激源性减弱。结论细胞因子鸡尾酒法是诱导DC成熟的最佳方法;而丁酸钠可以抑制DC成熟,改变DC的免疫状态。
Objective To investigate the immune effect of different cytokines on immature dendritic cells (DC) in vitro. Methods The human monocyte-derived DCs were induced in the presence of recombinant human GM-CSF (rhGM-CSF, 1000 U/ml) and interleukin (IL)-4 (500 U/ml). The effects of tumor necrosis factor (TNF)-α (1000 U/ml), lipopolysaccharide (LPS) (1 rag/L), the cocktail of cytokines (TNF-α, IL-6, IL-1β, PGE2 ), and sodium butyrate (1 mmol/L) on the DCs were detected by flow cytometry (FCM), endocytic activity, T cells stimulatory proliferation capacity, and IL-12 production. Resuits The cocktail of cytokines could up-regulate the major histocompatibility complex (MHC) class II and costimulatory molecules of DCs, decrease the endocytic activity 186. 74 ± 38.66, induce a stage of T- cell stimulation 18. 23 ± 2. 22 and promote the T helper cell type 1-skewing factor IL-12 production (656. 18 ±38.52) ng/L. But the sodium butyrate induced reverse result on DCs compared to cocktail of cytokines. Conclusion The most effective method for inducing dendritic cells maturation was cocktail of cytokines. The sodium bntyrate could inhibit the development of mature DCs, resulting in impaired DC function, and modify the outcome of the subsequent immune response.