大鼠调节性树突状细胞的培养与生物学特性鉴定  

Culture and biological characterization of rat regulatory dendritic cells

作　　者：高晓燕[1,2] 黄必军[2] 丘惠娟[1] 

机构地区：[1]中山大学肿瘤防治中心,广州510060 [2]华南肿瘤学国家重点实验室

出　　处：《中华实验外科杂志》2012年第11期2252-2254,共3页Chinese Journal of Experimental Surgery

摘　　要：目的培养大鼠调节性树突状细胞（rDC）并探讨其生物学特性。方法培养3种不同树突状细胞（DC）：（1）通过加入1×10^-6mol/L地塞米松（DXM）获得未成熟DC（imDC）；（2）通过加入1mg／L脂多糖（LPS）活化获得成熟（matDC）；（3）通过1×10^-6mol／LDXM预处理1mg／LLPS活化获得rDC。流式细胞仪检测各种DC表型，酶联免疫吸附试验（ELISA）法测量细胞因子水平。结果rDC表现为部分成熟的细胞表型，细胞表面CIM0表达较imDC组明显增加，CD86和人类主要组织相容性复合体（MHC）-Ⅱ表达与imDC类似。rDC分泌产生白细胞介素（IL）-12、IL-10水平都显著高于imDC，但IL-12生成上调在很大程度上被DXM预处理抵消，因此，反映了机体免疫反应状况的IL-10／IL-12比值较后者有显著升高（1．15±0．33、0．43±0．08；P〈0．05）。结论DXM预处理LPS活化可以诱导大鼠骨髓细胞获得rDC，表现为部分成熟的细胞表型，细胞表面CIM0表达及分泌1L-10／IL-12比值较imDC组增高。Objective To investigate the culture and biological characterization of rat regulatory dendritic ceils （rDCs）. Methods DCs were cultured for 9 days from the bone marrow （BM） -derived cells, supplemented with 20 ng/mL rGM-CSF and 8 μg/L rIL-4. Imature DCs （imDCs） were generated by adding 1 ×10^-6 mol/L DXM at day 6. To obtain rDCs, 1 mg/L lipopolysaceharide （LPS） was added at day 7 for activating imDCs after pretreatment with DXM. DCs phenotype was established by using flow cytometry. The production of proinflammatory cytokines was measured by using enzyme linked immunosorbent assay （ELISA）. Results The rDCs showed a partial mature phenotype, including a slight up-regulation of CD40 while low expression of MHC- H and CD86. The production of proinflammatory cytokines, in particu- lar IL-12, by rDCs was significantly less than that by matDCs, while rDCs and matDCs produced similar a- mounts of cytokine IL-10, leading to an increased IL-10/IL-12 ratio （ P 〈 0. 05 ） for rDCs. Conclusion The combination of GM-CSF/IL-4 and activation with LPS after pretreatment with DXM can induce DCs dif- ferentiation from BM-derived cells, which is a convenient and effective way for rDCs culture in vitro.

关 键 词：调节性树突状细胞 抗原递呈细胞 细胞因子 

分 类 号：R392[医药卫生—免疫学]