CpG ODN1826刺激人外周血树突状细胞对胃癌细胞杀伤效应的研究

目的探讨CpGODN1826在体外增强树突状细胞（dendriticcells,DC）抗胃癌作用的影响。方法分离正常人外周血DC,用GM-CSF和IL-4培养至第5天分组：GM-CSF＋IL-4组、GM-CSF＋IL-4＋TNF-α组、GM-CSF＋IL-4＋LPS组和GM-CSF＋IL-4＋CpGODN组。自外周血单个核细胞（PBMC）诱导分离DC,流式细胞仪检测其表面标志,MTT法测定其刺激同种异体淋巴细胞增殖的影响,检测其活性及对胃癌细胞的杀伤效应,ELISA检测各组分泌IL-12情况。结果体外培养第10天,CpGODN组出现大量典型的DC形态;流式细胞仪检测CpGODN组显著高表达CD40、CD1a、CD80、CD86、MHC-Ⅱ和分泌IL-12;在体外能强烈刺激同种异体混合淋巴细胞的增殖,显著增强DC杀伤MKN-45细胞的活性。结论 CpGODN可显著地诱导人外周血DC分化成熟,增强DC对胃癌细胞的杀伤作用。

The study on killing activity of human peripheral blood dendritic cells by CpG ODN1826 on MKN-45

Objective To explore whether CpG ODN1826 can affect the antitumor activity of dendritic cells （ DC） against gastric cancer in vitro. Methods DC was isolated from normal human peripheral blood by GM-CSF ＋ IL-4,GM-CSF ＋ IL-4 ＋ TNF-α,GM-CSF ＋ IL-4 ＋ LPS and GM-CSF ＋ IL-4 ＋ CpG ODN. The surface signal of DC was analysed by fluorescence-actived cell sorting（ FACS） ,and the abilities to stimulate proliferation of allogenic lymphocyte by DC and antitumor experiment were detected by MTT assay,and IL-12 released by every DC was detected by ELISA. Results A number of cells had typical morphology characteristics on the 10th day of CpG ODN group and LPS group,the CpG ODN group cells expressed the highest level of CD40,CDla,CD80,CD86,MHC-Ⅱand secretion of IL-12,and was potent to stimulate the proliferation of allogenic lymphocytes,and enhanced the antitumor activity of DC. Conclusions CpG ODN may evidently enhance the differentiation and maturation of DC and increase the antitumor capacity by stimulating PBMC in vitro.