抗原致敏DC与CIK共培养体外抗小细胞肺癌的研究-东南大学学报医学版

抗原致敏DC与CIK共培养体外抗小细胞肺癌的研究

单位：新疆医科大学附属肿瘤医院检验科, 新疆乌鲁木齐 830011

分类号：R-33;R734.2

出版年·卷·期（页码）：2017·36·第二期（225-229）

摘要：

目的：观察小细胞肺癌NCI-H209细胞株抗原致敏树突细胞（DC）与同源细胞因子诱导杀伤（CIK）细胞共培养后的DC-CIK细胞体外对小细胞肺癌NCI-H209的杀伤活性，为小细胞肺癌的特异性细胞免疫治疗提供实验依据。方法：将健康成人外周血单个核细胞来源的DC经NCI-H209细胞株抗原致敏后与同源CIK细胞共培养，实验分3组：NCI-H209抗原致敏DC与CIK共培养组（A组）、未致敏DC与CIK共培养组（B组）和单纯CIK组（C组）。流式细胞仪检测DC及CIK免疫表型，使用CCK-8法检测3组效应细胞对NCI-H209细胞的杀伤活性。结果：体外诱导培养出的抗原致敏DC和未致敏DC经流式检测，均高表达CD80、CD86、CD11c。CIK细胞随着培养天数的延长主要效应细胞CD3⁺CD8⁺、CD3⁺CD56⁺表达持续升高，至第14天流式检测结果示：A组和B组CD3⁺CD8⁺表型分别为71.8%和65.2%，C组CD3⁺CD56⁺达到31.1%。CCK-8法检测结果发现，相同效靶比下A组对NCI-H209细胞株的杀伤活性明显高于B、C组，且随着效靶比增加，其杀伤效应亦显著增加。效靶比为5：1时A组杀伤率为（68.82±2.55）%，B组杀伤率为（56.64±2.06）%，C组杀伤率为（55.98±2.63）%（P=0.007）；效靶比为10：1时A组杀伤率为（74.06±2.64）%，B组杀伤率为（60.11±2.74）%，C组杀伤率为（58.34±1.88）%（P=0.005）；效靶比为20：1时A组杀伤率为（79.17±3.51）%，B组杀伤率为（65.33±2.48）%，C组杀伤率为（60.47±2.85）%，（P=0.001）。结论：用NCI-H209细胞裂解物分离的混合蛋白致敏DC可以提高CIK细胞杀伤NCI-H209细胞效应，为小细胞肺癌的综合治疗提供新的策略。

Objective: To observe the dendritic cells(DC)-homologous cytokine induced killer(CIK) cells in vitro killing activity of small cell lung cancer NCI-H209 after the co-culture of the sensitized DC by the small cell lung cancer NCI-H209 cells antigen and CIK cells, provide the experimental basis for the treatment of small cell lung cancer of specific cellular immunity. Methods: The DC cells from healthy adult human peripheral blood mononuclear cells were sensitized by NCI-H209 cell line and co-cultured with CIK cells, the effector cells were divided into three groups: the sensitized DC by NCI-H209 antigen and CIK cells co-culture group(group A), the unsensitized DC and CIK cells co-culture group(group B), simple CIK group(group C). DC and CIK cells were detected by flow cytometry, and the cytotoxicity of three groups of effector cells to NCI-H209 cells was detected by CCK-8 assay. Results: In vitro culture of antigen-sensitized DC and unsensitized DC were detected by flow cytometry, high expressions of CD80, CD86, CD11c were found. CIK cells with prolongation of culture days, CD3⁺ CD8⁺, CD3⁺ CD56⁺ expressions of the main effector cells remained elevated, on the 14th day flow detection showed that CD3⁺ CD8⁺ phenotypes in the group A and group B were 71.8% and 65.2%, CD3⁺ CD56⁺ in the group C was 31.1%. By CCK-8 method, the results showed that group A was significantly higher than that of the group B and group C, and the killing effect of the group A was also significantly increased with the increase of the target ratio. When the effect target ratio was 5:1, the killing rate was(68.82±2.55)% in the group A, (56.64±2.06)% in the group B, (55.98±2.63)% in the group C(P=0.007); when the effect target ratio was 10:1, the killing rate was(74.06±2.64)% in the group A, (60.11±2.74)% in the group B, (58.34±1.88)% in the group C(P=0.005); when the effect target ratio was 20:1, the killing rate in the group A was(79.17±3.51) %, (65.33±2.48)% in the group B, (60.47±2.85)% in the group C(P=0.001). Conclusion: The sensitized DC by NCI-H209 can improve the killing effect of CIK cells, and provide a new strategy for the comprehensive treatment of small cell lung cancer.

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