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目的探讨复制缺损型腺病毒载体(Ad)介导异种AFP修饰的树突状细胞(DCs)诱发抗肝癌免疫、打破肿瘤免疫耐受的效果。方法从HepG2和Hepa16细胞中克隆人和小鼠AFP,插入Ad中构建AdhAFP和AdmAFP。用AdhAFP或AdmAFP感染小鼠骨髓来源的DC后,在无或有删除CD4或CD8情况下免疫C57BL/6小鼠,7d后取脾细胞行51Cr释放实验,检测特异性CTL杀伤活性;或给免疫小鼠接种Hepa16肝癌细胞,观察荷瘤小鼠成活情况。结果AdhAFP/DCs免疫小鼠1周后其细胞毒性T淋巴细胞杀伤活性明显强于AdmAFP/DCs。AdhAFP/DCs免疫小鼠后1周接种5×106Hepa16肝瘤细胞,2个月后仍然有80%的小鼠无瘤生长;而接种1×106Hepa16细胞至AdmAFP/DCs免疫小鼠,2个月后小鼠成活率为20%。删除小鼠CD4或CD8T细胞均使AdhAFP/DCs诱发的抗肿瘤免疫反应消失。结论Ad介导异种AFP修饰的DCs能有效地打破肿瘤的免疫耐受,诱发强烈的抗原特异性细胞免疫反应,这种特异性细胞免疫反应是CD4和CD8依赖性的。
Objective To investigate the effect of replication-deficient adenovirus vector (Ad) -mediated heterologous AFP-modified dendritic cells (DCs) in inducing anti-hepatocellular immunity and breaking tumor immune tolerance. Methods Human and mouse AFP were cloned from HepG2 and Hepa16 cells and inserted into Ad to construct AdhAFP and AdmAFP. After infection of mouse bone marrow-derived DCs with or without AdhAFP or AdmAFP, C57BL / 6 mice were immunized with or without CD4 or CD8 depletion, and splenocytes were harvested 51 days after 7 days to detect specific CTL-killing activity, or immunized Mice were inoculated with Hepa16 hepatoma cells to observe the survival of tumor-bearing mice. Results AdhAFP / DCs immunized mice after 1 week cytotoxic T lymphocyte cytotoxic activity was significantly stronger than AdmAFP / DCs. Immunized mice with AdhAFP / DCs were inoculated with 5 × 106Hepa16 hepatoma cells one week later, and 80% of mice remained tumor-free after 2 months. When mice were immunized with 1 × 106Hepa16 cells to AdmAFP / DCs for 2 months Mouse survival rate of 20%. The deletion of AdhAFP / DCs-induced antitumor immune response in mouse CD4 or CD8T cells disappeared. Conclusion Ad-mediated AFP-modified DCs can effectively break down the immune tolerance of tumor and induce a strong antigen-specific cellular immune response, which is CD4 and CD8 dependent.