HIV-1 Tat_(49-57)/HPV16 E7_(49-57)/pGM-CSF纳米颗粒疫苗抗肿瘤研究

The anti-tumor immunological effects of a novel self-assembled nanoparticle vaccine with HIV-1 Tat_(49-57)/HPV16 E7_(49-57) fusion peptide and GM-CSF DNA in mice

目的探讨HIV-1 Tat49-57-HPV16 E749-57CTL融合多肽与pORF9-mGMCSF质粒自组装成的纳米颗粒的抗肿瘤能力。方法合成Tat49-57-HPV16 E749-57融合肽,以富含正电荷的Tat49-57与表达mGM-CSF的DNA质粒通过静电引力作用自组装成纳米粒。凝胶阻滞实验、DNase I保护实验、透射电镜和Western blot等方法对纳米颗粒进行鉴定。构建预防性及治疗性肿瘤动物模型观察Tat-E7/pGM-CSF纳米颗粒疫苗在动物模型体内抗瘤效应。结果构建Tat-E7/pGM-CSF纳米颗粒,确定最适多肽/DNA电比r=2.0;当r=2.0时所制备的颗粒为类圆形,大小均匀一致,绝大部分颗粒直径均分布于20~80 nm之间。该纳米颗粒疫苗显著制了荷瘤小鼠的肿瘤生长,延长其存活时间,提高了存活率。结论 Tat-E7/pGM-CSF融合性纳米颗粒疫苗动物模型的在体抗肿研究证实了其具有良好效果。

This study aimed to design a novel peptide-based self-assembled nanoparticle HPV16 vaccine through combining cell-penetrating peptide HIV-1 Tat49-57 that was fused with HPV16 E749-57 CTL epitope and GMCSF gene,and investigate how it improves the immune response and the therapeutic outcome of anti-tumor ex vivo and in vivo.Nanoparticles were prepared and identified by transmission electron microscope,gel retardation and DNase I protection assays.The in vivo anti-tumor effectiveness of nanoparticle Tat-E7/pGM-CSF was investigated in the prophylactic and therapeutic mouse models.This type of vaccine formulation formed 20-80 nm nanoparticles at charge ratio 2.0,and this vaccine type was associated with decreased tumor growth and enhanced long-term survival in animals.These results suggest the nanoparticle Tat-E7/pGM-CSF designed in this study is a promising novel approach to enhance the potency of peptide-based cervical cancer vaccines.