单用A23187诱导外周血单个核细胞成树突状细胞及其介导耐药肿瘤细胞杀伤的实验研究  

作　　者：王宝中[1] 王丽萍[2] 王彦文[1] 张丽华[1] 孙桂明[1] 

机构地区：[1]聊城市人民医院肿瘤科,252000 [2]聊城市人民医院保健科,252000

出　　处：《当代医学》2010年第10期12-13,共2页Contemporary Medicine

摘　　要：目的探索一种快速、高效的由外周血单个核细胞(PBMC)获取树突细胞(DC)的方法,并探索该种来源DC对耐药肿瘤细胞的杀伤作用。方法通过单用A23187或联合细胞因子(GM-CSF、IL-4及TNF-a)将PBMC诱导分化为DC,观察细胞形态;流式细胞术检测诱导之DC表面分子表达;MTT法检测DC刺激异基因淋巴细胞增殖及其介导靶细胞的杀伤能力。结果上述两种细胞因子组合均能诱导PBMC向成熟DC分化,均高表达CD1a、CD80、HLA-DR、CD83和CD86;A23187组诱导72h获得DC数量高于细胞因子组的诱导率,也高于细胞因子组诱导7d的诱导率(P值均<0.05);两组所获DC在对异基因淋巴细胞增殖及介导T细胞对耐药肿瘤细胞的杀伤无显著差异(P值均>0.05)。结论单用A23187可快速(72h以内)诱导PBMC向成熟DC转化,且能介导杀伤耐药肿瘤细胞;与联合细胞因子相比,该方法是一种快速而高效的获取DCs的方法。Objective To explore a rapid and efficient way to generate dendritic cells from peripheral blood mononuclear cells （PBMC）, and to detect the cytotoxicity induced by these DCs to tumor cells of drug resistance. Methods PBMC were cultured with calcium ionphere A23187 alone or a DC differentiation cocktail consisting of GM-CSF, IL-4 and TNF-a, respectively. The expression of surface markers of induced DCs was analyzed by flowcytometry. The DCs stimulating the proliferation of allogenetic naive T cells and inducing cytotoxicity of T cells were determined by MTT assay. Results Microscopic examination revealed that under both the culture conditions PBMC became displaying DC morphology. At 72 hours in the culture systems containing A23187, there were higher proportions of cells with dendritic morphology, than that in the cocktail system （P〈0.05）. And the same did when cultured for 7 days, P〈0.05. In addition, both the groups of DCs were high expression of CD1a, CD80, HLA-DR, CD83 and CD86, and had no significant difference in allogenetic T cell proliferation response and induced T cell cytotoxicity to tumor ceils of drug resistance （P〉0.05）. Conclusion A23187 treatment is a rapid and efficient in vitro strategy for inducing dendritic cell from PBMC which can induce cytotoxicity to tumor cells of drug resistance.

关 键 词：A23187 MCF-7/ADM 树突状细胞 混合淋巴细胞反应 MTT试验 

分 类 号：R392[医药卫生—免疫学]