雷公藤红素对 CD1＋4单核细胞来源的树突细胞分化及成熟的影响

摘要

Objective To Make the differentiation and activation models of dendritic cells ( DCs ) and to investigate the effect of celastrol on the differentiation and maturation of human DCs in vitro .Methods Fresh human buffy coat was obtained , and peripheral blood mononuclear cells were isolated .Purified CDl4+monocytes were analyzed by FACSCalibur to confirm the purity of CD1+4 cells.The purified CD1+4 cells were cultured in complete 1640 medium supplemented with GM-CSF and IL-4 for 5 days and 7 days.For DC maturation, cells were stimulated with 5?g/mL lipopolysaccharide (LPS) at day 5.The control group was not added celastrol , but the celastrol group was added 10 and 20 nM celastrol, respectively. Then immature DCs ( iDC ) and mature DCs ( mDC ) were collected to detect the mean fluorescent intensity ( MFI ) of CD80 , CD83 , CD86 , HLA-DR and the absorb capacity of FITC-dextran by flow cytometry .Results Compared with the control group, the MFI of CD80 , CD83 , CD86 and HLA-DR of iDC and mDC in the celastrol group was lower , and the treat-ment of 20 nM was lower than the treatment of 10 nM, all P<0.05.The MFI of FITC-dextran in the negative control group (10.1 ±2.1) was the lowest, and was the highest in control group (243.7 ±31.3);in addition, the MFI of FITC-dextran in the 20 nM celastrol group (72.4 ±10.2) was lower than that of the 10 nM celastrol group (186.3 ±22.6), all P<0.05.Conclusions Celastrol inhibits CD1+4 monocytes differentiating into DCs , further inhibits phenotypic maturation of LPS-induced DCs , and suppresses the antigen-presenting function of iDC .%目的：制作免疫树突细胞（ DC）分化和激活模型，观察雷公藤红素对人DC体外分化和成熟的影响。方法采集浓缩人白细胞，淋巴细胞分离液分离单个核细胞，利用免疫磁珠法分离CD1＋4细胞，流式细胞分选仪确认分选纯度。将分选的CD1＋4细胞在含IL-4和GM-CSF的完全1640培养基中分别培养5 d（分化）和7 d（5 d后加入脂多糖刺激成熟）。对照组不添加雷公藤红素，雷公藤红素组分别加入10、20 nM的雷公藤红素。收集未成熟DC（ iDC）和成熟DC（ mDC），用流式细胞仪检测细胞表面标志CD80、CD83、CD86、HLA-DR平均荧光强度（ MFI）及细胞对FITC-dextran的摄取能力。结果雷公藤红素组iDC、mDC表面标记CD80、CD83、CD86及HLA-DR的MFI低于对照组，且雷公藤红素20 nM组低于10 nM组，P均＜0．05。阴性对照组、雷公藤红素10、20 nM组、对照组细胞摄取FITC-dextran的MFI分别为10．1±2．1、186．3±22．6、72．4±10．2、243．7±31．3，其中阴性对照组最低，对照组最高，雷公藤红素20 nM组低于10 nM组（P均＜0．05）。结论雷公藤红素可抑制CD1＋4单核细胞分化为DC，并抑制脂多糖诱导的DC表型成熟，还可抑制iDC的抗原提呈功能。