K562-DC融合瘤苗刺激的CIK／NK细胞在荷瘤NOD／SCID小鼠体内的杀瘤活性  

The cytotoxicity of CIK/NK cells stimulated by K562-DC fusion vaccines in NOD/SCID mice model for human erythroleukemia

作　　者：黎阳[1] 黄绍良[1] 张绪超[1] 方建培[1] 吴燕峰[1] 魏菁[1] 黄文革[1] 周敦华[1] 黄科[1] 林永潮[1] 

机构地区：[1]中山大学附属第二医院儿科,广州510520

出　　处：《中华血液学杂志》2008年第1期39-43,共5页Chinese Journal of Hematology

基　　金：国家自然科学基金（30500469）;卫生部临床重点基金（2004468）;广州科学仪器协作共用网基金（2006034）

摘　　要：目的探讨经K562细胞-树突细胞（DC）融合瘤苗刺激的脐血源性细胞因子诱导的杀伤（CIK）／自然杀伤（NK）细胞在荷瘤NOD／SCID小鼠体内的杀伤效能及其安全性。方法通过不同细胞因子的组合分别诱导脐血单个核细胞（MNC）成为DC和CIK／NK细胞，通过聚乙二醇（PEG）将灭活K562细胞与DC融合形成K562-DC融合瘤苗。以10：1比例在CIK／NK细胞培养体系中加入K562-DC融合瘤苗，制备K562-DC融合瘤苗刺激的CIK／NK细胞。小鼠均经尾静脉接种K562细胞1×10^6诱导荷瘤鼠；于接种肿瘤细胞后24h按不同分组分别经尾静脉输注K562-DC融合瘤苗刺激的CIK／NK细胞1×10^7、CIK／NK细胞1×10^7，同时设相应的非荷瘤鼠对照。比较各组小鼠的生存率、存活时间；用流式细胞术检测小鼠外周血、肝、肺组织中人CD13^＋细胞和外周血人CD56^＋细胞。结果在接种1×10^6 K562细胞后未经处理的小鼠39d内全部死亡，8只鼠中肉眼可见瘤块的小鼠5只，其中位于肝脏的4只、脾脏的1只。接种K562细胞后接受K562-DC瘤苗刺激的CIK／NK细胞治疗的8只小鼠第65天时死亡1只，抗瘤有效率为87．5％；接受CIK／NK细胞治疗小鼠死亡2只，时间分别为接种K562细胞后第56天及第62天，抗瘤有效率为75．0％，均未发现肉眼可见瘤块。该两组小鼠存活时间分别为（69．4±1．8）d、（67．2±5．3）d，两组间差异无统计学意义（P〉0．05），均高于未予治疗的荷瘤对照组[（30．4±4．6）d]（P〈0．01），荷瘤后接受K562-DC瘤苗刺激的和未经瘤苗刺激的CIK／NK细胞治疗的两组其余小鼠存活均超过70d。经CIK／NK细胞治疗的两组存活小鼠外周血，肝、肺组织中人CD13^＋细胞率差异无统计学意义，均显著低于荷瘤未治疗组（P〈0．01），而治疗的两组存活小鼠外周血人CD56^＋细胞率与输注不同CIK／NK细胞的非荷瘤对照组相比差异无统计学意义（P〉Objectives To study the in vivo efficacy and the safety of cord blood derived CIK/NK cells stimulated by K562-dendritic cells （DC） fusion vaccines in NOD/SCID mice model for human erythroleukemia. Methods DC and CIK/NK cells were both derived from cord blood mononuclear cells. DC were fused with inactivated K562 leukemia cell by PEG to produce K562-DC fusion vaccines. K562-DC fusion vaccines were co-cuhured with CIK/NK cells to prepare K562-DC fusion vaccine stimulated CIK/NK cells. NOD/SCID mice were inoculated with 1 ×10^6 K562 cells, 24 hours later, 1×10^7 vaccines stimulated CIK/ NK cells and 1×10^7 CIK/NK cells were transfused into the NOD/SCID mice. NOD/SCID mice without inoculation of K562 cells were used as control group. CD13 and CD56 positive cells were assayed by flow cytometry. Results All the leukemia NOD/SCID mice without therapy died within 39 days, tumor was found in 5 of 8 mice. One of 8 leukemia mice treated with K562-DC fusion vaccines stimulated CIK/NK cells died at the 65^th day, the anti-tumor response rate was 87.5%. Two of the leukemia mice treated with CIK/NK cells died at the 56^th and 65^th day respectively, the anti-tumor response rate was 75%. There was no significant difference in survival time between these two groups, and both survivals were longer than that of the control group. There was no significant difference in CD13 positive cells in the survival mice between these two groups, and both of that were less than that of the control mice. There was no significant difference in CD56 positive cells between the two treated groups and the control group. Conclusions Cord blood derived CIK/ NK cells stimulated by inactivated tumor cells retain the cytotoxicity and do not develop tumor in vivo.

关 键 词：K562细胞-树突细胞融合瘤苗 杀伤细胞 天然 NOD/SCID小鼠 

分 类 号：R686[医药卫生—骨科学]