乳腺癌细胞抗原和自身淋巴结树突状细胞对细胞因子诱导的杀伤细胞作用研究

The Research of the Effect of Breast Carcinoma Antigen and Dendritic Cell on the Cytokine Induced Killer

目的：从乳腺癌患者淋巴结 中分离和定向诱导培养扩增 树突状细胞（ＤＣ），观察其经自身肿瘤细胞抗原冲击后对自身细胞因子诱导的杀伤细胞（ＣＩＫ） 杀伤活性的影响。方法：取手术切除肿瘤周围转移的淋巴结，提取单个核 细 胞，将贴壁生长的细胞用细胞因子基因重组粒巨集落生长因子（ｒｈＧＭ－ＣＳＦ）、基因重组白介 素４（ｒｈＩＬ－４）、基因重组α－肿瘤坏死因子（ｒｈＴＮＦ－α）联合诱导培养ＤＣ，然后用 自身肿瘤细胞抗原冲击ＤＣ并作用ＣＩＫ，最后检测ＣＩＫ杀伤３种靶细胞（自身乳腺癌细胞、Ｋ５６２和 ＳＧＣ－７９０１细胞）的活性。结果：乳腺癌患者转移淋巴结中的贴壁细胞，经 细胞因子联合诱导培养，ＤＣ含量可达１５．３％～３７．０％。经自身肿瘤抗原冲击的淋巴结ＤＣ活化的Ｃ ＩＫ，杀伤自身肿瘤细胞活性达７１．３％，单纯ＣＩＫ为３７．０％，两者比较差异有显著性（Ｐ＜０ ．０１）；而对Ｋ５６２和ＳＧＣ－７９０１细胞杀伤活性无明显差异。结论：肿瘤周围转 移淋巴结贴壁细胞在体外能定向诱导扩增出大量ＤＣ；ＤＣ经自身肿瘤抗原冲击后能明显提高ＣＩ Ｋ对自 身肿瘤细胞的杀伤活性。

Objective: To Observe the effect of DC, which was assaulted by tumor cell antigen on the activity-killing of kill cell induced by cy tokine, and insolated cultured and amplified by orientally inducing from lymph n odes of the patients with breast carcinoma. Methods: We removed the ma tastatic lymph nodes around the tumor, extracted the mononuclear cells from whic h DC was induced and cultured combined with cytokines rhGM-CSF, rhIL-4, and rh TNF-α. The DCs through assault of the tumor cell antigen activated the CIK. Th e killing activity of CIK on three target cells (Breast Carcinoma, K562 and SGC -7901) was measured. Results: In the cultured flask after induction o f above cytokines, DCs raised to 15.3%-37.0%. The killing activity on tumor fro m CIK, which was activated by the DCs assaulted from tumor cell antigen was signi ficantly increased(73.1%). Meanwhile, the killing activity of CIK ,which was nor activated by DC, was significantly lower(37%) than that activated by DC(P<0 .01). There was no significant difference in killing activity of CIK on both K56 2 and SGC-7901 cells. Conclusion: The DCs can be induced and amp lified from the metastatic lymph nodes aroumd tumor. The killing activity of CIK , which was activated by the DCs assaulted from tumor cell antigen, increased. I t provides a theoretical basis for the tumor of immune treatment of DC.