SLE患者外周血DC的表面标志及其分泌IL-12和IFN-α的研究

Study on the surface markers on peripheral blood dendritic cells and their secretion of IL-12 and IFN-α in SLE patients

目的:分析SLE患者外周血中树突状细胞(DC)的表面标志,探讨DC和SLE发病之间的关系。方法:采用密度梯度离心法分离外周血单个核细胞(PBMC),在培养瓶中贴壁培养3h,吸去悬浮的细胞,联合应用GMCSF、IL4和TNFα刺激正常人及SLE患者外周血DC的增殖及分化成熟。用流式细胞仪分析DC的表面标志,用ELISA法检测培养9d的培养上清中IL12和IFNα的水平。结果:SLE患者DC表面CD1a、CD11c、CD40、CD83和CD123表达的阳性率,分别为:(58.88±7.64)%、(54.4±10.88)%、(37.29±8.08)%、(57.76±11.54)%和(13.14±4.44)%;健康志愿者(对照组)分别为:(47.71±4.01)%、(43.12±8.82)%、(28.59±7.07)%、(48.31±8.79)%和(9.85±3.97)%,两者相差明显(P<0.05)。SLE患者DC上CD80表达的阳性率为(55.16±10.12)%与对照组[(47.95±12.21)%]相差不明显(P>0.05)。培养上清中IL12的水平[(9.78±0.76)ng/L],较正常对照组[(7.49±0.74)ng/L]明显升高(P<0.05);SLE患者组IFNα的水平为[(2.95±0.61)ng/L]与对照组[(2.70±0.29)ng/L]相比升高不明显(P>0.05)。SLE患者非活动期与活动期培养上清中IL12和IFNα的水平无明显差异。结论:SLE患者的DC可能是通过其抗原递呈功能的增强及分泌IL12而参与本病的发病过程。

AIM: To analyze the surface markers on peripheral blood dendritic cells (DCs) in SLE patients and explore the relationship between the DCs and pathogenesis of SLE. METHODS: The peripheral blood monouclear cells (PBMCs) were separated by density gradient centrifugation. After culture of 3 hours in tissue culture flask, the suspended cells were removed and GM-CSF, IL-4 and TNF-α were used to stimulate the proliferation and maturation of the peripheral blood DCs from normal persons and SLE patients. The surface markers on the DCs were analyzed by flow cytometry and the levels of IL-12 and IFN-α in supernatants were measured by ELISA after culture of 9 days. RESULTS: The positive percentages of CD1a, CD11c, CD40, CD83 and CD123 expressed on DCs of SLE patients were (58.88±7.64)%, (54.4±10.88)%, (37.29± 8.08)% , (57.76±11.54)% and (13.14±4.44)%, respectively, whereas those of in normal subjects were (47.71± 4.01)% , (43.12±8.82)%, (28.59±7.07)%, (48.31± 8.79)% and (9.85±3.97)%, respectively, (P<0.05). But the positive proportion of CD80 expression was (55.16±10.12)% in SLE group and (47.95±12.21)% in the control group, without significant difference (P>0.05). The level of IL-12 in SLE group was (9.78±0.76) ng/L, higher than that in normal group. The level of IFN-α in SLE group(2.95±0.61) ng/L was not significant different from that in control group (2.70±0.29) ng/L (P>0.05). And there was no significant difference in IL-12 and IFN-α levels between non-active and active stages of SLE patients. CONCLUSION: The DCs may be involved in the pathogenetic process of SLE possibly by means of enhancement of antigen presenting function of DCs and secretion of IL-12.