多西环素对树突状细胞功能的影响

Regulation of Th1/Th2 Polarization by Doxycycline Via Modulating Dendritic Cells

目的探讨多西环素对树突状细胞的免疫表型和功能的影响。方法采集健康人的外周血,密度梯度离心法获得外周血单个核细胞,用rh IL-4和rh GM-CSF诱导分化为树突状细胞,通过LPS诱导树突状细胞成熟。树突状细胞诱导成熟过程中向培养基中分别加入0μg/m L、100μg/m L和300μg/m L的多西环素,通过流式细胞术检测树突状细胞表面HLA-DR、CD83和CD80的表达,用ELISA检测树突状细胞分泌TNF-α、IL-6、IL-12和IL-10,与T细胞共培养检测刺激同种异型T细胞的增值能力。结果与对照组相比,100 ng/m L和300 ng/m L多西环素处理组诱导树突状细胞表达HLA-DR表达下调。多西环素可下调树突状细胞分泌IL-1β、IL-12、TNF-α和IL-6,也可抑制树突状细胞刺激同种异型T细胞增殖,并呈剂量依赖性。结论多西环素作用于树突状细胞成熟过程,导致树突状细胞下调HLA-DR的表达,抑制树突状细胞分泌IL-1β、IL-6、IL-12和TNF-α,下调其刺激同种异型T细胞的增值能力。

Objective To explore the effect of doxycycline on the phenotypes and functions of dendritic cells. Methods PBMCs from healthy donors were generated using density gradient centrifugation. PBMCs were cultured in complete RPMI medium containing GM-CSF and IL-4 and were differentiated into DCs. DCs were divided into three groups and treated by LPS or LPS together with 100ng/mL or 300ng/mL doxycycline. The surface phenotype molecule, HLA-DR, CD83 and CD80, were determined by flow cytometry. Cytokines in supernatants were detected by ELISA. Re- sults HLA-DR expression of DCs treated with 100ng/mL and 300ng/mL doxycycline was down-regulated significantly compared to control group. Doxycycline treatment reduced the secretion of IL-1/3, IL-12, TNF- a and IL-6, and inhibit- ed allogeneic T cell proliferation by DCs in the dependence of dose. Conclusion In the DC maturation process, doxycy- cline lowered HLA-DR expression of DCs, and inhibited DC secretion of IL-1 /3, IL-6, IL-12 and TNF- a, and de- creased its ability to stimulate allogeneic T cells.