中文摘要：

目的探讨雷公藤甲素（TPT）对分泌IL-10的树突细胞（DC）亚群CD11clowCD45RBhighDC功能的影响。方法采用磁珠分选技术获得C57BL/6小鼠脾脏CD11clowCD45RBhighDC和CD4＋T淋巴细胞。CD11clowCD45RBhighDC分别加入1、10、20ng/ml TPT后进行培养,以不加TPT的细胞作为对照,流式细胞仪检测细胞表面分子CD40、CD80、CD86、I-a/e的表达,ELISA法检测培养液中IL-10的含量。将CD4＋T淋巴细胞分为正常对照组（未作任何处理）、未刺激组（与未经TPT处理的CD11clowCD45RBhighDC混合培养）、高浓度TPT刺激组（与经20ng/ml TPT处理后的CD11clowCD45RBhighDC混合培养）、高浓度TPT刺激＋抗体1（抗IL-10抗体）组、高浓度TPT刺激＋抗体2（IL-10的同型对照抗体）组,采用MTT法测定CD4＋T淋巴细胞的增殖活性,流式细胞仪检测CD4＋T淋巴细胞培养液中IL-4和IFN-γ的含量。结果与对照组相比,TPT能显著降低CD11clowCD45RBhighDC表面分子CD40和I-a/e的表达,增强CD86的表达及IL-10的分泌,其中IL-10的分泌水平随TPT浓度的增加而增加。高浓度TPT刺激组和高浓度TPT刺激＋抗体2组细胞增殖活性及IFN-γ分泌水平明显低于未刺激组,IL-4分泌水平明显高于未刺激组,而高浓度TPT刺激＋抗体1组细胞增殖活性及IFN-γ分泌水平明显高于未刺激组,IL-4分泌水平明显低于未刺激组。结论 TPT可促进CD11clowCD45RBhighDC IL-10的分泌,诱导CD4＋T淋巴细胞向Th2型分化,并可通过激活CD11clowCD45RBhighDC介导机体的免疫抑制活性。

英文摘要：

Objective To explore the influence oftriepoxide （TPT） on the function ofCDllc WCD4SRBhgh dendritic cell （DC） subset which produces IL-10. Methods Splenic CDllcIWCD4SRBh@ DCs and CD4＋ T cells were harvested from CS7BL/6 mice by magnetic bead sorting. CD 1 lctWCD4SRBhJgh DCs were treated with various doses of TPT （0, 1, 10, 20ng/ml）, and those without TPT as controls. Flow cytometry was used to determine the expression of CD1 lclWCD4SRBhgh DC surface molecules, including CD40, CD80, CD86, I-a/e, and IL-10 levels in CDI lcCD4SRBhgh DC culture supernatants were determined by sandwich enzyme-linked immunosorbent assays （ELISA）. The CD4 T cells were divided into the following S groups： control group, untreated group （TPT-untreated CD1 l cWCD4SRBhigh DC＋CD4＋ T cells）, high-dose TPT-treated group （20ng/ml TPT-treated CD1 lclWCD4SRBhSh DC＋CD4＋ T cells）, high-dose TPT-treated ＋ antibody I group （20ng/ml TPT-treated CD1 lcCD4SRBhsh DC＋CD4＋ T cells and IL-10 antibody） and high-dose TPT-treated ＋ antibody II group （20ng/ml TPT-treated CD1 lcWCD4SRBhgh DC＋CD4＋ T cells and IL-10 antibody isotype）. Proliferative activity of CD4＋ T lymphocytes was determined by MTT, whereas IL-4 and IFN- / contents in CD4＋ T cell culture supernatants were determined by flow cytometry. Results TPT markedly downregulated the expression of CD40 and I-a/e while upregulating the expression of CD86 on CDllcWCD45RBhsh DC surface and IL-10 secretion. The enhancement of IL-10 secretion level hinged upon the rise of TPT concentration. The proliferative activity and IFN- / secretion level in high-dose TPT-treated group and high-dose TPT-treated ＋ antibody lI group were markedly lower than those in the untreated group with significantly higher IL-4 secretion levels in TPT, whereas the proliferative activity and IFN- secretion level in high-close TPT-treated group and high-dose TPT-treated ＋ antibody I group were significantly higher than those in the untreated group with markedly lower IL-4 secr