梅毒螺旋体膜蛋白纳米核酸疫苗的优化及接种策略研究

项目名称： 梅毒螺旋体膜蛋白纳米核酸疫苗的优化及接种策略研究

项目编号： No.30800996

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 赵飞骏

作者单位： 南华大学

项目金额： 22万元

中文摘要： 本研究在筛选构建Tp膜蛋白Gpd单基因核酸疫苗在新西兰兔体内进行免疫活性初步研究的基础上，进一步构建Tp Gpd-IL-2基因融合壳聚糖纳米核酸疫苗；证实细胞因子佐剂hIL-2能否有效辅佐疫苗强化细胞免疫；同时探索验证核酸疫苗肌注初免-CpG ODN-蛋白黏膜免疫加强的疫苗接种策略能否全面激发机体免疫，特别是黏膜免疫；建立Tp感染的兔动物模型，确切评价该类疫苗的免疫效果。研究结果显示各Tp Gpd DNA疫苗组经肌肉注射免疫新西兰兔均可诱生较强的体液和细胞免疫应答。IL-2基因与Tp Gpd抗原基因不管是融合表达还是非融合表达均能显著增强TpGpd单基因疫苗的免疫效应和免疫保护作用。CS纳米颗粒包裹Tp Gpd DNA疫苗对疫苗的免疫效应及抗Tp皮肤感染有一定促进作用。CS纳米颗粒包裹及IL-2基因佐剂的联合应用能刺激pcD/Gpd 疫苗产生更强的免疫效应及更好的抗Tp皮肤感染效果。采用pcD/Gpd-IL-2疫苗肌注初免，CpGODN+ Gpd-IL-2蛋白鼻饲加强免疫的接种策略既能刺激机体产生较强的体液和细胞免疫效应，还能刺激较强的黏膜免疫效应，激发更有效的免疫保护作用。

中文关键词： 梅毒螺旋体;核酸疫苗;白细胞介素2;Gpd;膜蛋白

英文摘要： This study is based on the work that we have constructed the single-gene DNA vaccine and finished its study about immune activity, we construct a fusion DNA vaccine of Tp Gpd and interleukin 2 (IL-2) vectored by chitosan (CS) nanoparticles. It is then determined whether IL-2 as a molecular adjuvant and CS nanoparticles as vectors could enhance the immune response and protective efficacy of Tp Gpd vaccine in Tp-challenged rabbits.At the same time, we verify the Vaccination strategies that the Primary immunization of DNA vaccine be carried out by intramuscular injection and the Second immunization of recominant protein vaccine with CpG ODN be complied with mucosal immunization can enhance and overall stimulate the Immune effector, especially mucosal immunity. Conclusions ：Strong cellular immunity and humoral immunity responses could be induced by all Tp Gpd DNA vaccines in New Zealand Rabbits.Both pcD/Gpd-IL-2 vaccine and pcD/Gpd+pcD/-IL-2 vaccine could induce more powerful immune response and immune protection than pcD/Gpd vaccine. Immunization with Tp Gpd DNA vaccines wrapped with CS nanoparticles properly promote immune response and of the appropriate Gpd DNA vaccines.Immunization with pcD/Gpd-IL-2 vaccine by the Vaccination strategie of Primary immunization of pcD/Gpd-IL-2 vaccine carried out by intramuscular injection and the Second immunization of CpGODN with Gpd-IL-2 recombinant protein accine complied with mucosal immunization nasal immunization could induce a best immune response and lowest Ratios of Tp positive skin lesions(0%) and ulcer lesions(3.33%) .

英文关键词： Treponema pallidum；Outer membrane protein；nucleic acid vaccine；Gpd；interleukine 2