HPV16 E786-93纳米颗粒疫苗的免疫杀伤效果研究  

Study of Immune Killing Effect Induced by HPV16 E786-93 Nano-Vaccine

作　　者：胡张可 邓雄威[2] 赵佳琦 韩卢 石思伟 董晓筱 肖向茜[1] 盛望[1] HU Zhang-Ke;DENG Xiong-Wei;ZHAO Jia-Qi;HAN Lu;SHI Si-Wei;DONG Xiao-Xiao;XIAO Xiang-Qian;SHENG Wang(College of Life Science and Bioengineering,Beijing University of Technology,Beijing 100124;National Center for Nanoscience and Technology,Beijing 100190,China)

机构地区：[1]北京工业大学生命科学与生物工程学院,北京100124 [2]国家纳米科学中心,北京100190

出　　处：《生物技术通讯》2020年第2期135-141,共7页Letters in Biotechnology

基　　金：国家自然科学基金(31770999)。

摘　　要：目的:研究人乳头瘤病毒16型(HPV16)E786-93CTL表位肽与免疫佐剂CpG-ODN自组装纳米颗粒疫苗在肿瘤模型中的免疫效果。方法:化学合成HPV16 E786-93CTL表位肽,与CpG-ODN通过静电相互作用自我组装形成纳米颗粒,采用透射电镜、动态光散射等方法鉴定纳米颗粒的性质;构建治疗性肿瘤动物模型,观察纳米颗粒疫苗在动物模型体内的抗瘤效应;通过流式细胞术测定瘤内浸润性T细胞比例、外周血及脾脏内T细胞比例;体外培养小鼠骨髓来源树突状细胞(BMDC),测定经纳米颗粒刺激的BMDC培养上清中的IL-6和IL-12p40浓度。结果:构建了直径约210 nm大小均匀的纳米颗粒,该纳米颗粒疫苗能显著抑制荷瘤小鼠的肿瘤生长,且对于小鼠瘤内浸润性T淋巴细胞及脾脏内、外周血中的T淋巴细胞比例均有提升,同时能在体外有效刺激BMDC分泌细胞因子。结论:HPV16E786-93CTL表位肽联合CpG-ODN自组装纳米颗粒疫苗能有效杀伤肿瘤细胞,抑制肿瘤增长,改善体内免疫微环境。Objective:To study the immune effect of human papillomavirus type 16(HPV16)E786-93/CpG-ODN self-assembled nano-vaccine in tumor model.Methods:The HPV16 E786-93CTL epitope peptide was chemically synthesized and self-assembled with CpG-ODN by electrostatic interaction to form nanoparticles.The transmission electron microscopy and dynamic light scattering were used to characterize nanoparticles.Then the anti-tumor effect of nano-vaccines in therapeutic mouse models was observed.The proportion of infiltrating T lymphocytes in tumor,peripheral blood and spleen were determined by flow cytometry.Finally the concentrations of IL-6 and IL-12p40 in the culture supernatant of BMDC stimulated with nanoparticles were determined.Results:This type of vaccine formulation formed 210 nm nanoparticles.The nano-vaccine can significantly decreased tumor growth,and increased the tumor infiltrating T lymphocytes and the T lymphocytes in spleen and peripheral blood.At the same time,the nano-vaccine can effectively stimulate BMDC to secrete cytokines in vitro.Conclusion:HPV16 E786-93peptide-based nano-vaccine can effectively kill tumor cells,inhibit tumor growth,and improve the immune microenvironment in vivo.

关 键 词：人乳头瘤病毒16型(HPV16)E7 CPG-ODN 抑瘤效应 表位肽纳米疫苗 

分 类 号：R392.1[医药卫生—免疫学] R73-36[医药卫生—基础医学]