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目的 探讨来氟米特(LEF)处理前后系统性红斑狼疮(SLE)患者树突状细胞(DC)表面标志及功能的改变,揭示LEF治疗SLE的作用机制,为开展“抑制性DCs”治疗SLE奠定实验基础。方法 (1)分离SLE患者外周血单核细胞,用细胞因子诱导DC成熟, LEF组再加入A7717262(来氟米特的活性代谢产物)培养。第9天收集DC细胞,流式细胞仪检测CD80、CD83、CD86和HLA DR的表达。(2)分别将A771726处理或不处理的第9天DC和T细胞进行培养, 72h后用MTT法检测DC刺激淋巴细胞增殖的能力,FACS检测T细胞亚群和ELISA检测培养上清中IL 10和IFNγ水平。结果A771726处理后虽DC形态无改变,但DC表达CD83、CD86和HLA DR百分数较对照组均明显降低(72 70±1 77vs 79 36±4 80, 63 50±14 06vs. 83 91±9 81, 80 44±12 56vs. 90 51±8 63,P值均<0 01)。A771726处理后的DC,其刺激T细胞增殖相应的吸光度值明显降低,混合培养的上清液中IL 10水平较无A771726处理的DC与T细胞的混合培养上清液明显降低,而IFNγ两者间无显著差异;但见CD+4 CD+25CTLA+4 T细胞百分比增高。结论 LEF在体外可抑制SLE患者外周血DC的成熟;未成熟DC能抑制T细胞增殖及T细胞向Th2 细胞转化,诱导CD+4 CD+25CTLA+4 T细胞产生,从而纠正SLE患者的部分免疫紊乱。
Objective To investigate the changes of dendritic cells (DCs) surface markers and functions in patients with systemic lupus erythematosus (SLE) before and after leflunomide (LEF) treatment and to reveal the mechanism of action of LEF on SLE. Lay the foundation for the experiment. Methods (1) Peripheral blood mononuclear cells were isolated from patients with SLE, DCs were induced to mature by cytokines, and A7717262 (active metabolite of leflunomide) was added into LEF group. DCs were collected on the 9th day, and the expressions of CD80, CD83, CD86 and HLA DR were detected by flow cytometry. (2) DCs and T cells were treated with or without A771726 on day 9 respectively. After 72 hours, the ability of DCs to stimulate lymphocyte proliferation was detected by MTT assay. T cell subpopulations were detected by FACS and IL 10 And IFNγ levels. Results After treatment with A771726, the morphology of DCs was not changed, but the percentage of CD83, CD86 and HLA DR expression in DCs was significantly lower than that in control group (72 70 ± 1 77 vs 79 36 ± 4 80, 63 50 ± 14 06 vs 83 91 ± 9 81, 80 44 ± 12 56 vs. 90 51 ± 8 63, P <0.01). A771726-treated DCs significantly reduced the corresponding absorbance values for stimulating T cell proliferation, and the IL-10 levels in the mixed culture supernatants were significantly lower than those of the mixed culture supernatants of DCs and T-cells without A771726 treatment, whereas both IFNy There was no significant difference between the two groups; however, the percentage of CD + 4 CD + 25CTLA + 4 T cells was increased. Conclusions LEF can inhibit the maturation of peripheral blood DC in SLE patients in vitro. Immature DC can inhibit the proliferation of T cells and the transformation of T cells to Th2 cells, and induce the production of CD + 4 CD + 25 CTLA + 4 T cells to correct the partial immunity of SLE patients disorder.