摘要
目的:观察不同浓度三氧化二砷(ATO)对系统性红斑狼疮(SLE)患者外周血体外培养树突状细胞分化成熟和功能的影响,探讨其作用机制,初步阐明DC是否为ATO治疗SLE的作用靶点。方法:分离SLE患者外周血单个核细胞,用粒细胞巨噬细胞集落刺激因子(granulocyte macrophage colony stimulating factor,GM-CSF)、白细胞介素-4(interleukin-4,IL-4)细胞因子诱导DC成熟,加入不同浓度ATO培养。培养第9d收集DC细胞,流式细胞仪检测CD80、CD86和HLA-DR的表达。MTT法检测DC刺激淋巴细胞增殖的能力,ELISA法检测混和淋巴细胞反应培养上清IL-10和IFN-γ水平。结果:1.ATO处理后的DC表达CD80、CD86和HLA-DR百分数较对照组明显降低,P均<0.05,且随ATO浓度的增加DC表达CD80、CD86和HLA-DR百分数降低;2.ATO处理后的DC与T细胞混合培养,其刺激T细胞增殖相应的OD值较对照组明显降低,且随浓度增加降低越明显。3.其混合培养的上清液中IL-10水平较无ATO处理的DC与T细胞的混合培养上清液明显降低,P<0.05,而IFN-γ水平无统计学差异,P<0.05结论:ATO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,从而纠正SLE患者的部分免疫紊乱。
Objective：To detect the effects of different concentration of arsenic trioxide（ATO） on differention and maturation and function of peripheral blood dendritic cells（DCs） in SLE patients,and to explore its mechanism,clarifing whether the DC is the therapeutic targets to use ATO for SLE patients.Methods：The mononuclear cells were isolated from peripheral blood in SLE patients and developed into DCs by adding GM-CSF and IL-4.In ATO group,different concertrations of ATO was then added after adding the above cytokines.DCs were harvested after 9 days culture.CD80,CD86 and HLA-DR surface markers on DCs were detected by flow cytometry.MTT assay detects mixed lymphocyte reaction.IL-10 and IFN-γin the supernatant of MLR were detected by EL ISA. Results：1.The percentage of CD80 、CD86 and HLA-DR expression on the DCs treated with ATO were lower than those on the DCs without ATO-treated,all P0.05,and with the increasing of concertrations,the percentage of their reduction.2.The OD value related with T lymphocytes proliferation were lower in the MLR with the DCs treated with ATO than that in the MLR with the DCs without ATO treated,both P0.05,and with the increasing of concertrations,OD value was significantly lower.3.IL-10 level in the MLR supernatant of the group treated with ATO was lower than that in the control group,P0.05.but IFN-γlevel has no significant difference,P0.05）.Conclusion：ATO can inhibit DCs maturation,and then the immature DCs can also inhibit T cells proliferation and refrain T cells from dividing into Th2 subtype in SLE patients.
出处
《牡丹江医学院学报》
2010年第6期18-23,共6页
Journal of Mudanjiang Medical University
基金
广西自然科学基金(桂自科0991136)
广西青年科学基金(桂科青0542055)
广西卫生厅重点课题(桂卫重200829)
广西卫生厅自筹课题(桂卫Z2001048)资助项目
