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目的了解4-1BB配体(4-1BBL)治疗小鼠实验性肝癌时可能引起的负调控因素,以及可溶性PD-1(sPD 1)与4-1BBL协同治疗肿瘤的效应和机制。方法按照5×105／只的剂量将H22肝癌细胞接种于Balb／c小鼠右后腿肌肉内,建立小鼠实验性肝癌模型,随机分成5组,每组12只。空白对照组注射等渗盐水;pcDNA3．1组、p4-1BBL组和 pPD-1A组分别注射质粒pcDNA3．1、p4-1BBL和pPD-1A;p4-1BBL+pPD-1A组为联合治疗组,同时注射质粒p4- 1BBL和pPD-1A。观察小鼠肿瘤生长情况及存活率,检测两种基因在瘤周组织中的表达,对各组小鼠残存瘤细胞进行表型分析,检测肿瘤组织中的淋巴细胞。结果单独或联合转染4-1BBL基因和sPD-1基因均显示出抗肿瘤作用．尤以联合转染组小鼠肿瘤的生长抑制效果最为明显,该组有42％的小鼠肿瘤被完全抑制(其他各组完全抑制率为0);至实验第6周,联合转染组小鼠存活率达到100％,明显高于pcDNA3．1组(30％)、1)4-1BBL组(65％)、pPD-1A组(62％)。流式细胞术检测结果表明,p4-1BBL组残存瘤细胞上负调节性分子B7-H1和B7-DC的表达水平明显高于其他各组瘤细胞, 联合治疗组小鼠瘤周组织中CD8(?)T淋巴细胞数量明显增加。结论4-1BBL在促进抗肿瘤免疫应答的同时．也会引起负调节因素的上调,sPD-1可与4-1BBL产生协同治疗效应,增强机体抗肿瘤作用。
Objective To investigate the possible negative regulators of 4-1BB ligand (4-1BBL) treatment in mice with experimental hepatocarcinoma and the effect and mechanism of soluble PD-1 (sPD 1) in combination with 4-1BBL on tumor. Methods H22 hepatoma cells were inoculated into the muscle of right hind leg of Balb / c mice at a dose of 5 × 105 / mouse. Experimental mice model of liver cancer was established and divided randomly into 5 groups of 12 rats. The blank control group was injected with isotonic saline. The pcDNA3.1 group, the p4-1BBL group and the pPD-1A group were injected with pcDNA3.1, p4-1BBL and pPD-1A respectively. The p4-1BBL + pPD- Plasmids p4-1BBL and pPD-1A were injected simultaneously. The growth and survival rate of the mice were observed. The expression of the two genes in the peritumoral tissues was detected. The phenotypes of the remaining tumor cells in each group were analyzed, and the lymphocytes in the tumor tissues were detected. Results The 4-1BBL gene and sPD-1 gene alone or in combination showed antitumor effects. Especially in the combined transfection group, the growth inhibition effect of the tumor in the mice was most obvious. In the group, 42% of the mice were completely inhibited (the other groups were completely inhibited by 0); by the sixth week of the experiment, the combined transfection group was small The survival rate of mice reached 100%, significantly higher than pcDNA3.1 group (30%), 1) 4-1BBL group (65%), pPD-1A group (62%). The results of flow cytometry showed that the expression levels of B7-H1 and B7-DC on the remaining tumor cells in p4-1BBL group were significantly higher than those in other groups. CD8 (superscript + ?) The number of T lymphocytes increased significantly. Conclusion 4-1BBL can promote anti-tumor immune response at the same time. Can also cause negative regulation factors, sPD-1 and 4-1BBL produce synergistic therapeutic effect, enhance the body’s anti-tumor effect.