壳聚糖介导增强幽门螺杆菌Lpp20 DNA疫苗黏膜免疫效果的研究  

Chitosan-DNA nanoparticles enhancing the immune response of Helicobacter pylori Lpp20 DNA vaccine following the mucosal immunization

作　　者：曹斌[1] 张艳[1] 刘志杰[1] 于文[1] 余敏君[1] 

机构地区：[1]南华大学病原生物研究所,衡阳421001

出　　处：《中华微生物学和免疫学杂志》2011年第2期140-145,共6页Chinese Journal of Microbiology and Immunology

基　　金：湖南省自然科学基金课题（06JJ2093）;湖南省高校科技创新团队资助项目（湘教通[2010]212号）

摘　　要：目的 探讨壳聚糖包裹DNA疫苗黏膜免疫效果.方法 采用复凝聚法制备载幽门螺杆菌脂蛋白 Lpp20基因的壳聚糖（CS）纳米粒（NPs）,并对CS/DNA纳米粒的特质进行研究 用载基因壳聚糖纳米粒黏膜免疫（滴鼻和口服）小鼠,检测免疫小鼠的细胞和体液免疫水平.结果 裸质粒pcDNA3.1（＋）/Lpp20与CS/DNA NPs通过黏膜免疫均能诱导小鼠产生有效的免疫应答.CS/DNANPs滴鼻和口服免疫组诱导的抗体明显升高,与裸质粒组相比有明显差异（P＜0.05）,同时CS/DNANPs滴鼻和口服免疫组小鼠脾淋巴细胞经特异性抗原刺激后,刺激指数及培养上清中IFN-γ和IL-4含量明显高于裸质粒组、壳聚糖组和PBS组,且滴鼻免疫组高于口服组.结论 壳聚糖纳米粒能增强pcDNA3.1（＋）/Lpp20核酸疫苗的黏膜免疫（滴鼻和口服免疫）效果 载Lpp20基因壳聚糖纳米粒滴鼻免疫比口服免疫能诱导更强的细胞和体液免疫应答.Objective To investigate the immune response of mucosal immunization of new chitosan（CS） nanoparticles coating DNA vaccine. Methods The chitosan nanoparticles containing plasmid DNA encoding H. pylori lipoprotein Lpp20 gene were prepared using a complex coacervation method and then its speciality were analyzed. We then administered the naked plasmid DNA and chitosan-DNA nanoparticles to 6-week-old female BALB/c mice by intranasal or oral mucosal routes to observe the humoral and cellular immune responses. Results Naked plasmid pcDNA3.1 （ ＋ ）/Lpp20 and chitosan-pcDNA3. 1 （ ＋ ）/Lpp20 nanoparticles both induced effective immune response in mice through mucosal vaccination. Specific IgG and sIgA antibodies of chitosan-pcDNA3. 1 （ ＋ ）/Lpp20 nanoparticles groups were higher than that of naked pcDNA3.1 （ ＋ ）/Lpp20 group. The concentration of cytokines IFN-γ and IL-4 in cultural supernatant of T lymphocytes from chitosan-pcDNA3. 1 （ ＋ ）/Lpp20 nanoparticles immunized mice increased greatly than that of control groups. After stimulated by corresponding antigen, the stimulation index of intranasal or oral delivery of chitosan-pcDNA3. 1 （ ＋ ）/Lpp20 nanoparticles group was significantly higher than that of pcDNA3.1（ ＋ ）/Lpp20 group, CS group and PBS control group. Moreover, systemic and mucosal immune responses in mice induced by intranasal immunization were stronger than that of oral immunization. Conclusion Chitosan nanoparticles enhanced the immune response of pcDNA3.1 （ ＋ ）/Lpp20 DNA vaccine by intranasal or oral administration in BALB/c mice. Compared to oral administration, intranasal delivery of chitosan-pcDNA3.1 （ ＋ ）/Lpp20 DNA nanoparticles could induce stronger cellular and humoral immune responses in BALB/c mice.

关 键 词：壳聚糖纳米粒 幽门螺杆菌 LPP20 核酸疫苗 免疫原性 

分 类 号：R392[医药卫生—免疫学]