Tat-E7/pORF-mGM-CSF颗粒疫苗的特异性CTL效应研究  

Immunological effects of a novel self-assembled nanoparticle vaccine containing HIV-1 Tat_(49-57)/HPV16 E7_(49-57) fusion peptide and GM-CSF DNA

作　　者：张月[1] 玄秀云[2] 郝飞[3] 唐隽[1] 

机构地区：[1]蚌埠医学院附属解放军第105医院皮肤科,合肥230001 [2]山西医科大学微生物与免疫教研室,太原030001 [3]第三军医大学附属西南医院皮肤科,重庆400038

出　　处：《免疫学杂志》2017年第5期383-388,共6页Immunological Journal

基　　金：安徽省自然科学基金(1408085MH149);南京军区医学科技创新课题(ZX10)

摘　　要：目的前期制备并鉴定了Tat-E7/pORF-mGM-CSF颗粒疫苗,并在小鼠体内观察到良好的抗肿瘤效果,但其体内具体分子机制不清。本研究拟进一步利用该疫苗免疫小鼠,分析其体内激活T细胞应答的分子机制。方法通过免疫组化检测该纳米颗粒能否将外源基因转入哺乳动物细胞并在动物活体内正确表达;通过CCK-8淋巴细胞增殖实验、细胞内染色及流式细胞术(FCM)、ELISPOT实验、体外特异性细胞毒CTL-LDH释放实验分析疫苗诱导的短期及记忆性免疫应答和其介导的CTL毒性效应。结果当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;该纳米颗粒可将mGM-CSF基因导入哺乳动物细胞,并可在体外培养细胞和小鼠体内正确表达。该纳米颗粒疫苗在体外和体内均很大程度上提高了抗原表位特异性的免疫原性,可诱导T淋巴细胞增殖、分化,显著增强抗原特异性CTL应答及其产生的细胞毒效应。结论我们在本研究中设计并合成的Tat-E7/pGM-CSF纳米颗粒疫苗,通过体外免疫学研究证实了其具有良好效果,其机制可能是该颗粒疫苗的免疫可以维持高水平的特异性CD8^+记忆T细胞。We prepared and identified the Tat-E7/pORF-mGM-CSF nanoparticles vaccines previously,which has good antitumor effect in vaccined mice, but its specific mechanism is still unknown. In this study, we intend to find the molecular mechanism that how the vaccine induces T cell response. We detected the functions of inducting T lymphocyte proliferation, differentiation, antigen specific CTL production mediated cytotoxicity in vaccined mice. Homogeneous and dense particles with approximate diameters of 20-80 nm were observed by TEM.mGM-CSF could express in lymph nodes at the injection site and skin tissue in mice, which proved that the nanoparticles vaccine could carry exogenous genes into mammalian cells. The all results of proliferation assays,ELISPOT assays and intracellular cytokine staining and flow cytometry show that the vaccine could induce the effect of antigen specific CTL response. In conclusion, the Tat-E7/pGM-CSF vaccine can generate substantially improved epitope-specific immunity ex vivo and in vivo.

关 键 词：HPV 纳米颗粒 HIV Tat49-57 GM-CSF 

分 类 号：R392[医药卫生—免疫学]