目的：探讨自体树突状细胞（dendritic cell，DC）激活的细胞因子诱导的杀伤（cytokine-induced killer，CIK）细胞在晚期肾细胞癌（renal cell carcinoma，RCC）治疗中的临床效果与安全性。方法：采集22例2011年7月至2012年6月期间南京医科大学附属苏州医院肿瘤内科收治的22例RCC Ⅳ期患者\[男性12例、女性10例，中位年龄60.8岁（21~79岁）\]外周血单个核细胞（peripheral blood mononuclear cell，PBMC），体外制备成DC-CIK细胞。流式细胞术分析DC-CIK细胞中CD3+T、CD8+T、CD4+T、NK（CD3-CD56+）和NKT（CD3+CD56+）细胞的比例，MTT法检测DC-CIK细胞对白血病K562细胞（对NK细胞敏感）和肾癌786-0细胞（对NK细胞不敏感）的杀伤活性。受试患者于常规治疗（手术+化疗+放疗/细胞因子治疗）结束后4周进行DC-CIK细胞治疗，每次静脉回输细胞数约（5.0±0.5）×108个，5次为1疗程，共3个疗程，分别于疗程开始前与结束后1周内监测患者外周免疫学指标（淋巴亚群、细胞因子谱），严密观察并记录治疗过程中的不良反应。结果：DC-CIK细胞组成为CD3+细胞占（86.92±5.32）%、CD3+CD8+CD56+（NKT细胞）占（52.04±7.33）%、CD8-CD56+（NK细胞）占（785±315）%，DC-CIK细胞对786-0细胞和K562细胞的体外杀伤率相仿（效靶比为3∶1时，杀伤率分别为（16.5±1.7）%和（18.4±1.9）%，P=0.014）。患者接受DC-CIK细胞治疗后，外周血淋巴细胞亚群（CD3+T、CD4+T、CD8+T、CD3-CD56+NK细胞）比例无显著变化（P>0.05）；外周血中IL-2、IL-12和IFN-γ细胞因子水平较治疗前明显提升（均P＜0.05），而TNF-α 和IL-10变化不明显（均P＞0.05）。2例患者发生一过性发热，持续4~6 h恢复，1例出现短期乏力。结论：DC-CIK细胞体外能有效杀伤786-0细胞和K562细胞。 输注后可提升部分RCC患者免疫水平，安全性良好，可作为晚期RCC患者辅助治疗之一。

Objective : To investigate the clinical effect and safety of autologous dendritic cells (DCs) stimulated by cytokine-induced killer (CIK) cells in patients with advanced renal cell carcinoma (RCC). Methods: During July 2011 to June 2012, peripheral blood mononuclear cells (PBMCs) were collected from 22 patients (12 males and 10 females, median age 60.8 years \[21-79 years\]) with advanced renal carcinoma in the Department of Medical Oncology of Suzhou Hospital affiliated to Nanjing Medical University, and DC-CIK cells were individually prepared from these PBMCs. Flow cytometry was performed to analyz the proportion of CD3+T, CD8+T, CD4+T, NK（CD3-CD56+）and NKT（CD3+CD56+）cells in the DC-CIK cells. The cytotoxicity of DC-CIK cells on leukemia K562 cells (sensitive to NK cells) and renal carcinoma 786-0 cells (insensitive to NK cells) was detected by MTT assay. DC-CIK cells therapy was started when the conventional therapy (surgery+chemotherapy+radiotherapy/ cytokine therapy) finished and continued for 4 weeks. Each time, about (5.0±0.5)×108 DC-CIK cells were returned through intravenous transfusion, and each patient received 5 times (one cycle) of DC-CIK cells intravenous infusion. The immunological index such as lymphocyte subsets and cytokine concentration in the peripheral blood were detected before and after DC-CIK infusion. The adverse reactions during the treatment were closely observed and recorded. Results: In the total number of DC-CIK cells, the proportion of CD3 positive (CD3+) lymphocytes was (86.92±5.32)%, while the proportions of CD3+CD56+NKT cells and CD3-CD56+ NK cells were (52.04±7.33)% and(7.85±3.15)% respectively. DC-CIK cells showed similar in vitro killing activities on 786-0 cells and K562 cells in vitro, with a killing rate of (16.5±1.7) % and (18.4±1.9) % respectively (P=0.014), when the ratio of effect cells and target cells was 3 ∶1. After patients received DC-CIK cell therapy, the proportion of peripheral lymphocyte sub-group (CD3+T, CD4+T, CD8+T and CD3-CD56+NK cells) showed no significant change (P>0.05); compared to the cytokine levels prior to DC-CIK cells infusion, the levels of interleukine 2 (IL-2), interleukin 12 (IL-12) and interferon gamma (IFN-r) increased significantly (P<0.05), while the levels of tumor necrosis factor-α (TNF-α) and interleukine 10 (IL-10) changed unremarkably (P>0.05). Two patients experienced a transient fever lasting 4-6 hours and one felt a short time of fatigue after DC-CIK cells infusion. Conclusion: DC-CIK cells can kill 786-0 cells and K562 cells effectively. The DC-CIK cells infusion may improve immune response of some patients with a safe level, and it can be one of the assistant treatment methods for advanced RCC patients.

苏州市科技计划资助项目（No.SS0524)