PEDF对小鼠树突细胞表型及免疫功能影响的实验研究

Effects of PEDF on phenotypic and immunologic function of dentritic cells

目的探讨色素上皮衍生因子(PEDF)对小鼠骨髓源性树突细胞(BMDCs)表型及其免疫功能的影响。方法从小鼠骨髓中分离单个核细胞(MNC),用小鼠重组粒细胞巨噬细胞集落刺激因子(rmGM-CSF)、小鼠重组白细胞介素-4(rmIL-4)培养5d后将其分为5组,实验组分别加入50、100、200ng/mL PEDF,阳性对照中加1μg/mL脂多糖(LPS),阴性对照中加等量的RPMI1640培养基(对照组),继续培养3d。流式细胞术(FCM)分析DCs表面CD11c、CD80、CD86表达情况,混合淋巴细胞反应(MLR)检测BMDCs对T细胞的刺激能力,ELISA法检测其培养上清液中IL-12水平。结果 PEDF处理后的BMDCs明显上调了CD11c、CD80和CD86的表达,并能明显增强T细胞的活性,促进其分泌IL-12。结论 PEDF可明显上调体外培养的小鼠BMDCs免疫标记分子的表达,并增强其免疫活性。

Objective To explore the effects of pigmentary epithelium derived factor （PDEF） on the phenotypic and immunologic function of murine-derived dentritic cells（BMDCs） .Methods Mononuclear cells（MNCs） isolated from murine bone marrow were cultured in RPMI1640 medium containing rmGM-CSF and rmIL-4 for 5 d ,and were divided into five groups .MNCs were stimulated for 3 d with either 50 ,100 ,200ng/mL PEDF ,1 μg/mL LPS（positive control） or RPMI1640（negative control） .The expression of CD11c ,CD80 and CD86 on DCs surface were analyzed by the fluorescence activated cell sorting （FCM ） .The ability of PEDF-induced BMDCs to stimulated T cell maturation were determined by the CCK-8 method and the level of IL-12 in the culture supernatant was detected by ELISA .Results The PEDF-treated BMDCs expressed high levels of CD11c ,CD80 and CD86 ,enhanced the immunolog-ical activities of T lymphocyte and its secretion of IL-12 when compared with untreated DCs .Conclusion PEDF can significantly up-regulate the expression of DCs immunological labelled molecule in in vitro cultured murine and increase its immunological com-petence .