摘要
目的:探讨经黄芪多糖诱生脐血来源的树突状细胞(DCs)体外介导的抗白血病细胞的细胞毒效应。方法:无菌条件下采集脐血,用淋巴细胞分离液分离获得脐血单个核细胞。分为两组,实验组:在含有浓度100mg/L黄芪多糖的10%胎牛血清的RPMI-1640完全培养液中培养;对照组:RPMI-1640完全培养液中培养。①在培养过程中用倒置光学显微镜和透射电镜观察细胞形态学变化;FCM技术检测培养第12天两组细胞的细胞免疫表型。②MTT比色法检测DCs对同种异体T细胞的增殖作用。③MTT比色法检测由DCs介导的对K562细胞的抑制作用。结果:①在培养的第72h后实验组细胞形态开始变化,随着培养时间的延长,树突状结构更加明显,第12天细胞呈典型的树突状细胞形态;对照组细胞生长缓慢,培养至第12天细胞呈梭形巨噬细胞形态。培养至10d的实验组细胞透射电镜下呈典型的树突状细胞形态。培养12天实验组细胞高表达DCs特异性抗原CD1a、CD80、CD83和CD86,与对照组对应比较差异均有显著性(P<0.01)。②在混合淋巴细胞反应中,经黄芪多糖诱生的DCs具有明显激发同种异体T淋巴细胞增殖的能力,且随数量的增加而作用增强。③经黄芪多糖诱生的DCs负载肿瘤抗原后可诱导肿瘤特异性CTL产生抗白血病细胞毒效应,而对照单独T细胞对K562细胞的杀伤作用很弱,各对应效靶比实验组和对照组之间相比有显著的统计学意义(P<0.05或0.01);各实验组间比较差异显著(P<0.05或0.01);各对照组之间比较无统计学意义(P>0.05)。结论:①黄芪多糖可诱导脐血来源的单核细胞(即DCs前体细胞)定向分化为功能性(即成熟)DCs;②黄芪多糖诱生的DCs具有强烈激发同种异体T淋巴细胞增殖的能力,且随数量的增加而作用增强;③黄芪多糖诱生脐血来源的DCs可负载肿瘤抗原诱导特异性CTL的产生,发挥显著的抗白血病细胞毒效应。
Objective ： To investigate the anti - leukemia effect mediated by DCs derived from the cord blood monocyte induced by the APS. Methods： （1）The cord blood monocytes were isolated and the mononuclear cells were prepared from cord blood with Ficoll - hypaque centrifugation method, then cultured in the RPMI - 1640 or cultured in the RPMI - 1640 with APS （concentration 100mg/L） respectively. The morphotype of DCs was identified by inverted optical microscope or transmission electron microscope. The phenotype （CD1 a, CD80, CD86, and CD83 ） of cultured 12 days DCs was identified by flow cytometry. （2）The capability of stimulating allo - lymphocyte proliferation was tested with mixed leukocyte reaction （MLR） by MTT. （3）The inhibitory action to 1（562 cells by MTT. Results： （1）Cultured for 72 hours , The morphous of cell of the experiment group began to change from round to irregularity, appearing rough cell face and barb pustute. The longer cell cultured, the more obvious the dendritic structure is. The cell cultured for 12 days had the most typical dendritic structure. The control group cell had no change and became the macrophage when cultured for 12 days. The ex- periment group cell cultured for 10 days showed typical dendritic morphotype by TEM. The experiment group cell cultured for 12 days significantly expressed the high level phenotype of DCs（ CD1 a, CD50, CD86, and CD83） by flowcytometry. The difference has the significance when compared with the control group （ P 〈 0.01 ）. （2）The capability of stimulating lymphocyte proliferation of the cord blood monocyte derived dendritic cells was enhanced markedly in APS group. With the amount of DCs going up,the capability of stimulation becomes stronger. （3）The DCs induced by APS charged tumor anti- gen can induce Tumor - CTL specifically to generate anti - leukemia effect, while there was no anti - leukemia effect in the T cells, as the control group. Especially, when the ratio of the reaction cells and target cells was 100：1, K562 cells was suppressed obviously. Conclusion ：（1） APS could induce the cord blood monocyte into functional DCs.（2）APS induced DCs can strongly stimulate the allo -lymphocyte proliferation and with the amount of DCs going up,the capability of stim- ulation becomes stronger. （3）The DCs induced by APS charged tumor antigen can induce Tumor - CTL specifically and can generate marked anti -leukemia effect.
出处
《中华中医药学刊》
CAS
2009年第11期2360-2365,共6页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省中医药管理局科研基金资助项目(2005C009)
杭州市卫生局科研基金资助项目(2004A008)
关键词
脐血
树突状细胞
黄芪多糖
药理学
白血病细胞
免疫治疗
cord blood
DCs
astragalus polysaccharides
pharmacology
leukemia cell
immunotherapy
