Ren Wei, Wang Zhihua
Cancer Research Institute of Harbin Medical University, Harbin 150081, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Aug;24(8):1373-80.
To investigate the immunologic characteristics and cytotoxicity of the RetroNectin-activated cytokine-induced killer cells (CIK) against drug-resistant lung cancer cell lines DDP-A549 (DDP: Cisplatin). Peripheral blood mononuclear cells (PBMC) were collected from healthy donors and divided into two groups: group I and group II. Seeded samples of group I into culture flask precoated with RetroNectin and CD3MAb to induce the CIK cells while seeded the group II into culture flask precoated with CD3MAb. In both groups, IFN-gamma was put into the flask on the same day and then IL-2 on the second day. The proliferation of CIK cells was tested by cytometirc analysis. The cytotoxicity activity of CIK cells was determined by MTT assays. The phenotype changes of CIK cells were identified by flow cytometric analysis. Scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to view the cytotoxicity against DDP-A549 of CIK cells and the changes of DDP-A549. The total CIK cells significantly increased by 524.77 fold in cell proliferation number due to the activation to CIK cells of RetroNectin. The expression rate of CD3+CD56+ cells was (31.40 +/- 1.91)%. The cytotoxicity of CIK cells showed statistically significance between DDP-A549 and the sensitive strains of parental generation A549 (P < 0.01). There was no significant difference of CIK cells' cytotoxicity between two groups when the effector: target ratio was fixed (P > 0.05). RetroNectin can significantly improve the proliferation activity of CIK cells. There was no evident influence to the cytotoxicity of CIK cells. CIK cells may be used as the immuotherapy to lung adenocarcinoma owing to its significant inhibition to the proliferation of DDP-A549.
探讨RetroNectin激活的细胞因子诱导的杀伤细胞（CIK）对耐药肺癌细胞系DDP-A549（DDP：顺铂）的免疫特性和细胞毒性。从健康供体采集外周血单个核细胞（PBMC），分为两组：I组和II组。将I组样本接种到预先包被RetroNectin和CD3单克隆抗体的培养瓶中诱导CIK细胞，而将II组接种到预先包被CD3单克隆抗体的培养瓶中。两组均在同一天向培养瓶中加入γ干扰素，第二天加入白细胞介素-2。通过细胞计数分析检测CIK细胞的增殖情况。通过MTT法测定CIK细胞的细胞毒性活性。通过流式细胞术分析鉴定CIK细胞的表型变化。使用扫描电子显微镜（SEM）和透射电子显微镜（TEM）观察CIK细胞对DDP-A549的细胞毒性以及DDP-A549的变化。由于RetroNectin对CIK细胞的激活作用，CIK细胞总数在细胞增殖数量上显著增加了524.77倍。CD3 + CD56 +细胞的表达率为（31.40±1.91）%。CIK细胞对DDP-A549的细胞毒性与亲代A549敏感株之间具有统计学意义（P < 0.01）。当效应细胞：靶细胞比例固定时，两组CIK细胞的细胞毒性无显著差异（P > 0.05）。RetroNectin可显著提高CIK细胞的增殖活性。对CIK细胞的细胞毒性无明显影响。CIK细胞因其对DDP-A549增殖的显著抑制作用，可作为肺腺癌的免疫治疗方法。
