树突细胞与细胞因子诱导的杀伤细胞共培养增强其体内外抗肿瘤活性  

Coculture of dendritic cell with-induced killer results in a significant increase in cytotoxic activity of CIK to tumor cells in vitro and in vivo

作　　者：葛薇[1,2] 李长虹[1,2] 张伟[1,2] 韩钦[1,2] 邓为民[1,2] 陈磊[1,2] 尤胜国[1,2] 赵春华[1,2] 

机构地区：[1]中国医学科学院,中国协和医科大学血液学研究所血液病医院,天津300020 [2]中国医学科学院实验血液学国家重点实验室

出　　处：《中华血液学杂志》2004年第5期277-280,共4页Chinese Journal of Hematology

基　　金：卫生部科技专项基金资助项目 (wkz 2 0 0 0 1 3 4) ;天津市科技发展计划资助项目 ( 0 13 11112 1)

摘　　要：目的探讨与树突细胞 (DC)共培养能否增强正常人细胞因子诱导的杀伤细胞 (CIK)的体内外抗瘤活性。方法　分别按照常规方法从正常人外周血单个核细胞诱导DC和CIK细胞 ,将NB4白血病细胞冻融物 (LCL)冲击或未冲击的DC与CIK细胞共培养 (LCL DC +CIK、DC +CIK) ,以CIK细胞单独培养作为对照。用流式细胞术分析细胞表型 ,酶联免疫斑点法 (ELISPOT)测定分泌IFN γ的细胞数 ,51 Cr释放实验测定体外细胞毒活性 ,同时用NB4细胞系建立荷瘤裸鼠模型研究其体内抗肿瘤活性和归巢情况。结果　在培养第 1 5天 ,LCL DC +CIK与CIK细胞单独培养相比 ,增殖速率明显提高 [(1 8.2± 2 .1 )倍vs(1 1 .6± 2 .3)倍 ,P <0 .0 5 ],CD3 + CD56+ 表达水平也明显提高 [(5 1 .0 5± 2 .6 3) %vs(30 .1 8± 1 .4 5 ) % ,P <0 .0 5 ],分泌IFN γ的细胞数量明显增高 [(86 .33± 5 .5 1 ) / 1 0 4细胞vs(4 4 .6 1± 3.0 5 ) / 1 0 4细胞 ,P <0 .0 5 ],同时LCL DC +CIK对NB4、K5 6 2、KG1a的体外细胞毒活性增强。体内实验显示与单独培养CIK细胞相比 ,LCL DC +CIK细胞共培养后 ,可明显抑制接种瘤细胞裸鼠的成瘤率 ,提高裸鼠的长期无瘤存活率 (1 0 0 %vs 6 6 .7% ,P <0 .0 5 ) ,以DiI标记的LCL DC +CIK细胞在接种后 7d内可在脾脏、淋巴结及肿瘤?Objective To explore whether coculture of dendritic cells (DC) with cytokine induced killer (CIK) lead to an increase of cytotoxicity against tumor cells in vitro and in vivo. Methods DC and CIK were prepared from human peripheral blood mononuclear cells (PBMC) by conventional methods, the DC pulsed with or without NB4 leukemia cell lyses (LCL)was cocultured with the CIK (LCL DC+CIK and DC+CIK), CIK was used as control. Cells phenotypes were analyzed by flow cytometry, secretion of IFN γ was determined by ELISPOT assay, and cytotoxicity was assyed in vitro with 51 Cr release assay. A human leukemia cell NB4 bearing nude mice model was established to test in vivo antitumor efficacy and cell homing. Results Compared with CIK, LCL DC+CIK got a significant increasing of proliferation rate \[(18.2±2.1)times [WTBX]vs (11.6±2.3) times, P<0.05\] and CD 3 +CD 56 + expression rate \ and CD 3 +CD 56 + expression rate \[(51.05±2.63)% [WTBX]vs (30.18±1.45)% , P<0.05\], and the number of IFN γ secreting cells was increased significantly \, and the number of IFN γ secreting cells was increased significantly \[(13.86±3.28)/10 4 cells [WTBX]vs (8.74±2.53)/10 4 cells, n=12, P<0.05\].Meanwhile , LCL DC+CIK led to an increase of cytotoxic activity to NB4, K562, and KG1a cells, and showed significant inhibition of the growth of transplanted tumor cells and increased tumor free survival rate of nude mice (100% .Meanwhile , LCL DC+CIK led to an increase of cytotoxic activity to NB4, K562, and KG1a cells, and showed significant inhibition of the growth of transplanted tumor cells and increased tumor free survival rate of nude mice (100% vs 66.7%, P<0.05), DiI labeled LCL DC+CIK were detected in spleen, lymph node and tumor within a week after injection. There was no significant different in antitumor activity between LCL DC+CIK cell and DC+CIK cell. Conclusion Coculture of CIK with DCs can promote the efffect of CIK against tumor in vitro and in vivo. DC CIK is

关 键 词：树突细胞 细胞因子 杀伤细胞 抗肿瘤活性 DC CIK 细胞株 

分 类 号：R73-36[医药卫生—肿瘤]