日本血吸虫双价DNA疫苗的构建及其免疫保护性研究  

Construction and Expression of Schistosoma japonicum Bivalent DNA Vaccines and Their Efficiencies of Protective Immunity

作　　者：胡媛[1] 石佑恩[1] 朱晓华[1] 宁长修[1] 朱红刚[1] 

机构地区：[1]华中科技大学同济医学院病原生物学系,武汉430030

出　　处：《中国寄生虫学与寄生虫病杂志》2006年第2期97-101,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家高技术研究发展计划(863计划)资助项目(No.2004AA2-Z3212)~~

摘　　要：目的研究防治日本血吸虫病的双价DNA疫苗的免疫保护效果。方法构建共表达双价DNA疫苗pVI-VO2-mcs-SjFABP-Sj23和pVIVO2-mcs-Sj23-SjFABP,并用BamHI/EcoRI和BspHI/AvRⅡ进行双酶切和测序鉴定。通过免疫荧光法(IFAT)检测质粒在BALB/c小鼠骨骼肌细胞的表达。70只小鼠随机分为7组,每组10只,分别肌肉注射生理盐水、pVIVO2-mcs、pVIVO2-mcs-Sj23、pVIVO2-mcs-SjFABP、pVIVO2-mcs-Sj23-SjFABP、pVIVO2-mcs-SjFABP-Sj23和pVIVO2-mcs-Sj23-SjFABP+多糖佐剂,免疫1次。免疫后4周用40±2条尾蚴攻击感染,感染后45d剖杀,计数各组小鼠成虫数和肝虫卵数。结果双酶切反应和测序分析证明成功构建双价质粒DNA。IFAT结果提示双价质粒DNA可在小鼠骨骼肌细胞胞膜和胞浆中表达。双价DNA疫苗免疫小鼠后获得41.2%～53.8%的减虫率和47.0%～53.8%肝减卵率;pVIVO2-Sj23-SjFABP+多糖佐剂组获得68.9%的减虫率和84.0%肝减卵率,保护力显著高于单价疫苗和双价疫苗(P<0.05﹚。结论共表达的双价DNA疫苗在小鼠体内可产生较好的抗血吸虫感染的免疫保护效果。多糖佐剂组保护效果明显高于单价和双价疫苗组。Objective To study the efficiency of protective immunity afforded by the constructed bivalent DNA vaccines of Schistosoma japonicum. Methods The plasmids pVIVO2-mcs-SjFABP-Sj23 and pVIVO2-mcs-Sj23-SjFABP, co-expressed bivalent DNA vaccines, were constructed and identified. The presence of bivalent DNA vaccine in the mouse muscle cells was also tested by indirect immunofluorescent antibody tests （IFAT）. 70 BALB/c mice were randomly divided into seven groups to be injected with normal saline, pVIVO2-mcs, pVIVO2-mcs-Sj23, pVIVO2-mcs-SjFABP, pVIVO2-mcs- Sj23-SjFABP, pVIVO2-mcs-SjFABP-Sj23 plasmid DNA, and a mixture of pVIVO2-mcs-Sj23-SjFABP plasmid DNA and amylose adjuvant, respectively. At day 45 after challenge the mice were sacrificed. The number of adult worms and hepatic eggs were counted. Result Successful construction of co-expressed bivalent DNA vaccines were identified by restriction analysis and sequencing. It was confirmed by IFAT that the bivalent DNA vaccine was expressed in the plasma and on the surface of muscle cells from mouse. The worm reduction rate was 41.20%-53.85% and the egg reduction rate was 47.02%-53.83% bivalent DNA groups. Furthermore, the worm and egg reduction rates in pVIVO2-mcs- Sj23-SjFABP plasmid DNA and amylose adjuvant group were 68.89% and 84.04% respectively, significantly higher than single antigenic DNA vaccine and bivalent DNA vaccine （P〈0.05）. Conclusion The co-expressed bivalent DNA vaccines of Schistosoma japonicum can induce immune protection in mice. The protective immunity of amylose adjuvant group is higher than that of the univalent DNA and bivalent vaccines.

关 键 词：日本血吸虫 SJ23 SjFABP DNA疫苗 佐剂 

分 类 号：R392.11[医药卫生—免疫学] R532.21[医药卫生—基础医学]