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目的 探讨氧化修饰低密度脂蛋白 (oxidized lowdensitylipoprotein ,oxLDL)和低密度脂蛋白 (lowdensi tylipoprotein ,LDL)对人单核细胞源的树突状细胞 (monocytederiveddendriticcells,MDCs)免疫功能的影响。 方法 采用免疫磁珠法分离人外周血CD14 + 单核细胞 ,经含rhGM CSF(10 0ng/mL)和rhIL - 4 (2 0ng/mL)的Cell gro培养 ,使其分化为MDCs。分别以PBS和LPS(5 0 0 μg/mL)作阴性和阳性 (成熟 )对照 ,观察经5 0 μg/mL的oxLDL和LDL干预 4 8h后 ,流式细胞术检测MDCs表型 (CD1a、CD4 0、CD86、HLA DR) ,混合T淋巴细胞反应检测MDCs对淋巴细胞增殖的影响 ,FITC Dextran检测MDCs吞噬功能 ,ELISA检测细胞培养上清细胞因子 (IL -12、IL - 10和TNF α)浓度。结果 与PBS相比 ,5 0 μg/mLoxLDL干预的MDCs,不但可明显上调共刺激分子CD86的高表达 ,而且反映MDCs成熟程度的CD1a也表达增强 ,与此同时经oxLDL处理的MDCs吞噬作用却明显减弱 ,进一步的混合T淋巴细胞反应显示经过oxLDL刺激的MDCs促T细胞增殖作用明显增强 (P <0 .0 5 ) ;此外 ,oxLDL可明显促进MDC分泌细胞因子TNF α、IL - 10和IL - 12 (P <0 .0 5 ) ,但明显弱于LPS组 ,而LDL无此作用。结论 oxLDL在促进DCs成熟的同时 ,DCs激活T淋巴细胞的能力也明显增
Objective To investigate the effects of oxidized low density lipoprotein (oxLDL) and low density lipoprotein (LDL) on the immune function of human monocyte derived dendritic cells (MDCs). Methods Human peripheral blood mononuclear cells (CD14 +) were isolated by immunomagnetic beads method and cultured in Cell Gro containing rhGM CSF (10 0 ng / mL) and rhIL - 4 (20 ng / mL) to differentiate into MDCs. The negative and positive (mature) controls were treated with PBS and LPS (500 μg / mL) respectively. After 48 h intervention with 50 μg / mL oxLDL and LDL, the phenotypes of MDCs (CD1a and CD4 0, CD86 and HLA DR). The mixed lymphocyte reaction was used to detect the effect of MDCs on lymphocyte proliferation. The phagocytosis of MDC was detected by FITC Dextran. The levels of cytokines (IL - 12, IL - 10 and TNFα) . Results Compared with PBS, MDS induced by 50 μg / mL oxLDL not only significantly up-regulated the expression of costimulatory molecule CD86, but also enhanced the expression of CD1a, which reflects the maturation of MDCs. Meanwhile, phagocytosis of MDCs treated with oxLDL was obvious The further mixed T lymphocyte reaction showed that oxLDL - stimulated proliferation of T cells stimulated by oxLDL was significantly enhanced (P <0.05). In addition, oxLDL could significantly promote the secretion of cytokines TNFα, IL - 10 and IL - 12 (P <0.05), but significantly weaker than LPS group, but no such effect of LDL. Conclusions OxLDL can promote the maturation of DCs and the ability of DCs to activate T lymphocytes is also significantly increased