CpG ODN对人胃癌BGC823细胞RNA转染的脐血树突状细胞诱导CTL活性影响的研究  

Human cord blood dendritic cells with gastric carcinoma BGC 823 cells RNA transfection induce CTL antitumor activity by enhanced though CpG-ODN

作　　者：杜英[1] 黄玉敏[1] 孙蕾[1] 董子明[2] 

机构地区：[1]郑州大学基础医学院微生物学与免疫学教研室,河南郑州450052 [2]郑州大学基础医学院病理生理学教研室,河南郑州450052

出　　处：《中国现代医学杂志》2007年第11期1312-1315,1319,共5页China Journal of Modern Medicine

基　　金：河南省重大科技攻关项目(No:001170209)

摘　　要：目的探讨肿瘤细胞RNA转染脐血树突状细胞对CTL特异性抗肿瘤作用的影响,以及CpG ODN对脐血DC的免疫佐剂作用。方法分离脐血单个核细胞,经SCF、GM-CSF和IL-4诱导和BGC823RNA转染形成成熟DC,加入CpG ODN,通过形态学、表面标志和DC诱导T细胞增殖和CTL杀伤活性判断CpG ODN对DC功能的影响。结果肿瘤RNA转染后DC高表达MHC-I、MHC-II和CD54、CD80和CD86,对T细胞的促增殖作用和CTL杀伤活性显著增强,P<0.01。CpG ODN能显著促进DC功能。结论从脐血中诱导出DC,经肿瘤细胞RNA转染,可使DC表达MHC分子、协同刺激分子、黏附分子以及活化T细胞能力显著增强,CpG ODN具有增强DC抗原提呈功能的作用。[Objective] To investigate the human cord blood dendritic cells with gastric carcinoma BGC 823 cells RNA transfection induce CTL antitumor activity by enhanced through CpG-ODN. [Methods] The RNA of BGC823 cells were prepared by Trizol reagent. The cord blood monocytes were cultured to produced DCs with recombinant human stem cell factor （rhSCF）, recombinant human granulocyte-macrophage colony-stimulating factor （rhGM-CSF） and recombinant human interleukin-4 （rhIL-4）. The DCs were collected and tumor cell total RNA was transfected and CpG ODN was added in DC culture system. MHC-Ⅰ, MHC-Ⅱ and co-st/mulatory molecules on the surface of DCs were analyzed by FCM. The Mixed lymphocyte reation （MLR） and cytotoxicity of CTLs to BGC823 cells was assayed by MTT colorimetry. [Results] DCs transfected with total tumor ribonucleic acid （RNA） exhibited more MHC- Ⅰ, MHC-Ⅱ, CD54, CD 80 and CD86 than before DCs transfection, P 〈0.01. Mature DC with tumor RNA transfection induced superior tumor cytolytic activities compared with that immature DC. CpG ODN can enhance significantly that the DC activity, there are different comparing with control. [Conclusion] DCs can be obtained in vitro from human cord blood monocytes and express highly MHC-Ⅰ, MHC-Ⅱ and co-stimulatory molecules, and present tumor antigen to T cells and enhance the cytotoxicity of CTLs. Tumor RNA transfection and using CpG ODN is better methods in DC mature process and induction on CTL antitumor activity.

关 键 词：树突状细胞 BGC823 细胞毒活性 CpG ODN 脐血 

分 类 号：R735.2[医药卫生—肿瘤]