脐血CD3AK细胞体外抗肿瘤活性的实验研究

目的　探讨抗ＣＤ３ 单克隆抗体（ＡｎｔｉＣＤ３ＭｃＡｂ）＋ 重组人白细胞介素２（ｒＩＬ２） 激活的脐血杀伤细胞（ＣＤ３ＡＫ） 细胞对白血病细胞株Ｋ５６２ 、ＨＬ６０ 的杀伤作用。方法　采用间接玫瑰花结法测定脐血单个核细胞表面分化抗原，ＥＬＩＳＡ双抗夹心法测定肿瘤坏死因子α（ＴＮＦα）、ＩＬ６ 和ＩＬ８ ，用β液闪仪测定３Ｈ胸腺嘧啶核苷标记的靶细胞杀伤活性。结果　①经ＡｎｔｉＣＤ３ＭｃＡｂ＋ ｒＩＬ２ 激活的脐血单个核细胞免疫表型发生明显变化；②细胞因子水平较未激活组明显增高；③脐血ＣＤ３ＡＫ 细胞对Ｋ５６２、ＨＬ６０细胞均有杀伤作用，最佳杀伤条件：脐血单个核细胞浓度为１ ×１０６／ｍｌ，ＡｎｔｉＣＤ３ＭｃＡｂ １ μｇ／ｍｌ，ｒＩＬ２ １０００ Ｕ／ｍｌ，作用时间７２ 小时，效靶比１００∶１。结论　脐血单个核细胞能被ＡｎｔｉＣＤ３ ＭｃＡｂ＋ ｒＩＬ２ 激活为杀伤细胞，激活后有较强的细胞毒作用，且对白血病细胞株有明显的杀伤作用，该研究为应用活化脐血进行肿瘤的过继免疫治疗提供理论依据。

Killing of leukemic cells by Anti CD_3McAb plus rIL 2 activated cord blood

OBJECTIVE: To investigate the killing efficacy of cord blood monocyte activated by Anti-CD3 monoclonal antibody(McAb) in K562 and HL-60 cell lines. METHODS: By using indirect rosette forming test, antigen expression rate of monocyte was detected. The levels of interleukin-8 (IL-8), IL-6 and tumor necrosis factor alpha (TNF-alpha) were measured by ELISA. The killing efficacy in 3H-thymidine target cells was detected by beta-liquid scintillator. RESULTS: After being activated by Anti-CD3McAb and rIL-2, the antigen expression of monocyte was changed markedly. The levels of IL-8, IL-6 and TNF-alpha were increased; The Anti-CD3 McAb activated killer cells produced significant lysis in K562 and HL-60 cell lines and the optimum conditions were as follows: concentration of cells, 1 x 10(6)/ml; Anti-CD3McAb, 1 microgram/ml; rIL-2 1,000 U/ml; time of culture, 72 hrs; ratio of effector/target: 100:1. CONCLUSION: Anti-CD3 McAb activated killer cells exhibited a strong cytotoxicity to leukemic cells. Our study provides a theoretical basis for the adoptive immunotherapy of leukemia by activated cord blood.