Wang Yuliang, Wang Yinlong, Mu Hong, Liu Tao, Chen Xiaobo, Shen Zhongyang
Department of Clinical Laboratory Medicine, Tianjin First Central Hospital, Key Laboratory for Critical Care Medicine of the Ministry of Health, Tianjin 300192, P.R. China.
Department of Hernia and Abdominal Wall Surgery, Union Medicine Center, Tianjin 300121, P.R. China.
Mol Med Rep. 2015 May;11(5):3361-7. doi: 10.3892/mmr.2015.3239. Epub 2015 Jan 22.
Dendritic cell (DC)‑based cancer immunotherapy requires an immunogenic tumor‑associated antigen and an effective therapeutic strategy. Glypican 3 (GPC3) is a valuable diagnostic marker and a potential therapeutic target in hepatocellular carcinoma (HCC). The present study investigated whether DCs transduced with the GPC3 gene (DCs‑GPC3) and co‑cultured with autologous cytokine‑induced killer cells (CIKs) may induce a marked specific immune response against GPC3‑expressing HCC cells in vitro and in vivo. Human DCs were transfected with a green fluorescent protein plasmid with GPC3 by nucleofection and then co‑cultured with autologous CIKs. Flow cytometry was used to measure the phenotypes of DCs and CIKs. The co‑cultured cells were harvested and incubated with HCC cells and the cytotoxicity of the CIKs was assessed by nonradioactive cytotoxicity assay. The anti-tumor activity of these effector cells was further evaluated using a nude mouse tumor model. The results demonstrated that DCs‑GPC3 significantly promoted the autologous CIKs differentiation, as well as anti‑tumor cytokine interferon‑γ secretion. In addition, DCs‑GPC3‑CIKs significantly enhanced the cytotoxic activity against GPC3‑expressing HepG2 cells, indicating a GPC3‑specific marked immune response against HCC cells. The in vivo data indicated that DCs‑GPC3‑CIKs exhibited significant HepG2 cell‑induced tumor growth inhibition in nude mice. The results of the present study provided a new insight into the design of personalizing adoptive immunotherapy for GPC3‑expressing HCC cells.
基于树突状细胞(DC)的癌症免疫疗法需要一种具有免疫原性的肿瘤相关抗原和一种有效的治疗策略。磷脂酰肌醇蛋白聚糖3(GPC3)是肝细胞癌(HCC)中有价值的诊断标志物和潜在的治疗靶点。本研究调查了用GPC3基因转导的DC(DC-GPC3)与自体细胞因子诱导的杀伤细胞(CIK)共培养是否可以在体外和体内诱导针对表达GPC3的HCC细胞的显著特异性免疫反应。通过核转染用携带GPC3的绿色荧光蛋白质粒转染人DC,然后与自体CIK共培养。流式细胞术用于检测DC和CIK的表型。收获共培养的细胞并与HCC细胞孵育,通过非放射性细胞毒性试验评估CIK的细胞毒性。使用裸鼠肿瘤模型进一步评估这些效应细胞的抗肿瘤活性。结果表明,DC-GPC3显著促进自体CIK的分化以及抗肿瘤细胞因子干扰素-γ的分泌。此外,DC-GPC3-CIK显著增强了对表达GPC3的HepG2细胞的细胞毒活性,表明对HCC细胞有针对GPC3的显著特异性免疫反应。体内数据表明,DC-GPC3-CIK在裸鼠中对HepG2细胞诱导的肿瘤生长具有显著抑制作用。本研究结果为针对表达GPC3的HCC细胞的个体化过继性免疫疗法的设计提供了新的见解。