摘要
本研究探讨藏红花素(crocin)对白血病患儿骨髓树突状细胞(DC)体外增殖及功能的影响。采用Ficoll-Hypaque法分离急性白血病患者骨髓单个核细胞,实验分为6组。A组为空白对照组;B组加入终浓度为1.25mg/ml藏红花素;C组加入rhGM-CSF 75 ng/ml、rhIL-4 75 ng/ml、rhTNF-α50 ng/ml;D,E,F组分别加入与C组等量的细胞因子及藏红花素0.3125,1.25,5.0 mg/ml。培养至第9天,在倒置显微镜下观察各组细胞形态,对各组细胞进行计数;应用流式细胞仪检测各组细胞免疫表型;将DC与儿童急性白血病同种异体外周血淋巴细胞混合反应5d后观察淋巴细胞的增殖情况。结果表明,对照组及实验各组均得到一定数量典型的DC,实验各组细胞数目明显高于对照组,差异有显著统计学意义(P<0.01)。培养至第9天,流式细胞仪免疫分析显示C、D、E、F各组CD1a+、CD83+、HLA-DR+细胞比例明显高于A组(P<0.01),B组与A组之间比较差异无统计学意义(P>0.05)。混合淋巴细胞培养5 d后,随着刺激细胞的增加,A组及B组T细胞的刺激指数无明显增加(P>0.05);C组及E组T细胞的刺激指数明显增加,2组间差异有统计学显著意义(P<0.01)。而各组在刺激细胞数量一定的情况下,E组细胞刺激指数明显增加(P<0.01)。结论:藏红花素单用促进DC增殖的能力明显弱于与细胞因子的联合作用,但能协同细胞因子促进DC成熟。藏红花素诱导后的DC能促进T细胞增殖。
Abstract This study was purposed to investigate the effect of crocin on the proliferation in vitro and immune func- tion of dendritic cells （DC） derived from the bone marrow of children with acute leukemia. The mononuclear cells were isolated from bone marrow of leukemia children by Ficoll-Hypaque. The experiment was divided into six groups： blank control group （ A）, crocin 1.25 mg/ml group （ B ）, cytokines （ rhGM-CSF 75 ng/ml ~ rhIL-4 75 ng/ml ＋ rhTNF-et 50 ng/ml） group （ C）, cytokines ＋ crocin 0. 3125,1. 25 or 5.0 mg/ml groups （ D, E, F）. The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture. The DC of different groups were mixed with T cells just separated from peripheral blood of another children with acute lymphoblastic leukemia, and cultured with rhIL-2 200 U/ml for 5 d. The function of DC was detected by mixed lymphocyte reaction （MLR）. The results indicated that the test groups and control group all obtained a certain amount of typical DC, but the DC numbers in test groups were all higher than those in control group（P 〈0.01 ）. Cultured for 9 days, the rates of CDla~ ,CD83 ~ , and HLA-DR~ in group C,D,E,F were higher than group A（p 〈0.01 ）. There was no statistically significant difference between A and B groups （P 〉 0.05）. MLR showed that with the increasing of DC, the stimulation index of T cells in group A and B was not rising（ P 〉 0.05 ） ; the stimulated index of T cells in group C and E was significantly rising, there was statistically significant difference between them（ P 〈0.01 ）. When the number of stimulated cells was the same, the stimulation index of T cell in group E was the highest （P 〈0.01 ）. It is concluded that the capability of DC proliferationpromoted by crocin alone is lower than that of its combination with rhGM-CSF, rhlL-4 and rhTNF-a, but the crocin can synergically promote the maturity of DC cooperating with rhGM-CSF, rhlL-4 and rhTNF-a. The DC induced by crocin can particularly enhance the proliferation of T cells.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2012年第1期57-61,共5页
Journal of Experimental Hematology
