Wu Hao, Chen Bin, Peng Baogang
Department of General Surgery, Guangzhou Twelfth People's Hospital, Guangzhou, Guangdong 510620, P.R. China.
Department of Hepatobiliary Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China.
Exp Ther Med. 2018 Feb;15(2):1914-1917. doi: 10.3892/etm.2017.5633. Epub 2017 Dec 14.
This study investigated the effects of intratumoral injection of immunoactivator after microwave ablation on antitumor immunity in a mouse model of hepatocellular carcinoma. Hepatocellular carcinoma cell line Hepa1-6 was subcutaneously injected into C57/B6 mice to establish a mouse model of hepatocellular carcinoma. When tumor diameter reached 8 mm, microwave ablation was performed for 3 min with temperature controlled at 55°C. Cytokine sustained-release microspheres (CytoMPS) containing human interleukin-2 (hIL-2) and mouse granulocyte macrophage colony-stimulating factor (mGM-CSF) were injected into the tumor of mice in the experimental group (n=5) at 3, 7 and 14 days after ablation, while sustained-release microspheres containing no cytokine were used in the control group (n=5). Mice were sacrificed on the 17th day after ablation, and CD4 and CD8 T cells in peripheral blood were counted by flow cytometry. Spleen was collected from the mice to isolate lymphocytes. Lactate dehydrogenase (LDH) release assay was used to determine the cytotoxicity of spleen cells to Hepal-6 cells. Injection of CytoMPS after ablation increased the percentage of CD4 and CD8 T cells in peripheral blood. Cytotoxicity of CD8 CTL to Hepal-6 is significantly higher in experimental group than in control group (P<0.01). The results showed that intratumoral injection of CytoMPS containing hIL-2 and mGM-CSF can significantly increase the proportion of CD4 and CD8 T cells in blood and increase the cytotoxicity of CTL cells to tumor cells in mice with hepatocellular carcinoma.
本研究在小鼠肝细胞癌模型中,探讨了微波消融后瘤内注射免疫激活剂对抗肿瘤免疫的影响。将肝癌细胞系Hepa1-6皮下注射到C57/B6小鼠体内,建立小鼠肝细胞癌模型。当肿瘤直径达到8 mm时,进行微波消融3分钟,温度控制在55℃。在消融后第3、7和14天,将含有人类白细胞介素-2(hIL-2)和小鼠粒细胞巨噬细胞集落刺激因子(mGM-CSF)的细胞因子缓释微球(CytoMPS)注射到实验组(n=5)小鼠的肿瘤中,而对照组(n=5)使用不含细胞因子的缓释微球。在消融后第17天处死小鼠,通过流式细胞术计数外周血中的CD4和CD8 T细胞。从小鼠体内采集脾脏以分离淋巴细胞。采用乳酸脱氢酶(LDH)释放试验来测定脾细胞对Hepal-6细胞的细胞毒性。消融后注射CytoMPS可增加外周血中CD4和CD8 T细胞的百分比。实验组中CD8 CTL对Hepal-6的细胞毒性显著高于对照组(P<0.01)。结果表明,瘤内注射含有hIL-2和mGM-CSF的CytoMPS可显著增加肝癌小鼠血液中CD4和CD8 T细胞的比例,并增强CTL细胞对肿瘤细胞的细胞毒性。