类风湿性关节炎中树突状细胞亚群分析

Analysis of dendritic cell subsets in rheumatoid arthritis

目的分析类风湿性关节炎患者外周血和关节液中的DC(dendritic cell,DC)亚群以及DC亚群与炎症产生的关系。方法收集来自西南医院体检中心26例健康人外周血作为对照组,来自西南医院风湿免疫科28例RA患者的外周血及滑膜液作为实验组,其中RA患者的外周血和滑膜液来自同一RA患者。采用人外周血淋巴细胞分离液密度梯度离心法分离提取对照组和实验组外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)和滑膜液单个核细胞(synovial fluid mononuclear cells,SFMCs),流式细胞术检测如下DC亚群:浆细胞样DC(CD123^(+)DC)、髓系DC(CD11c^(+)DC),进一步分析CD11c^(+)DC亚群,CD1c^(+)DC和CD141^(+)DC。结果首先我们分析比较健康人外周血、RA患者外周血和RA患者滑膜液中各DC亚群,结果发现:健康人外周血CD123^(+)DC亚群的比例明显高于RA患者外周血(P<0.0001),而RA患者外周血和滑膜液CD123^(+)DC亚群的比例无显著性差异;而RA患者滑膜液中CD11c^(+)DC亚群占比例明显高于RA患者外周血(P<0.01);进一步对CD11c^(+)的亚群进行分析,结果发现RA患者滑膜液中CD1c^(+)DC所占比例明显高于RA患者外周血(P<0.05);而RA患者滑膜液中CD141^(+)DC亚群所占比例也显著高于RA患者外周血(P<0.001);最后我们分析了RA患者滑膜液中CD11c^(+)、CD1c^(+)以及CD141^(+)DC亚群与RA患者滑膜液中活化CD4^(+)CD45RO^(+)T细胞相关性,结果发现:CD1c^(+)DC的比例与CD4^(+)CD45RO^(+)T细胞比例有显著正相关(P<0.05)。结论RA患者滑膜液中CD11^(+)DC、CD1c^(+)DC和CD141^(+)DC亚群的比例明显高于患者外周血,提示mDC可能在RA的局部炎症中起着比较重要的作用。

This study was performed to investigate the percentage of dendritic cell(DC)subsets in the peripheral blood and synovial fluid of patients with rheumatoid arthritis(RA)and the relationship between DC subsets and local inflammation.We collected the peripheral blood of 26 healthy donors as a control group,and the peripheral blood and synovial fluid of 28 RA patients as experimental groups.Peripheral blood mononuclear cells(PBMCs)and synovial fluid mononuclear cells(SFMCs)were separated by the density gradient centrifugation using lympohprepTM.DC subsets plasma cell-like DC(CD123^(+)),myeloid DC(CD11c^(+)CD1c^(+)and CD11c^(+)CD141^(+))were analyzed by flow cytometry.Data showed that the percentage of CD123^(+)DC in peripheral blood of the healthy individual was significantly higher than that in the RA patient(P<0.0001),while there no significant difference in the percentage of CD 123+DC in peripheral blood and synovial fluid of RA patients.As to RA patients,the percentage of CD11c^(+)DC in synovial fluid was significantly higher than that of peripheral blood(P<0.01).Further analysis of CD11c^(+)DC subsets showed that the percentages of not only CD1c^(+)DC in synovial fluid of RA patients was significantly higher than that of peripheral blood(P<0.05),and the percentage of CD141^(+)DC was also significantly higher than that of peripheral blood(P<0.001).Correlation analysis showed that the percentage of CD1c^(+)DC and CD4+CD45 RO+T cell were positively correlated(P<0.05).Taken together,the percentage of CD11c^(+)DC,CD1c^(+)DC and CD141^(+)DC in synovial fluid of RA patients were significantly higher than that in peripheral blood,and mDC may play an important role in RA local inflammation.