血吸虫硫氧还蛋白免疫原性研究 Ⅱ日本血吸虫硫氧还蛋白DNA疫苗小鼠保护性免疫研究  

Studies on immunogenicity of Schistosoma thioredoxin Ⅱ. Study on protective effect of Schistosoma japonicum thioredoxin DNA vaccine against challenging infection in mice

作　　者：曹建平[1] 徐馀信[1] 韩海勃[1] 宋光承[1] 李小红[1] 卢潍媛[1] 沈玉娟[1] 刘述先[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室世界卫生组织疟疾血吸虫病和丝虫病合作中心,上海200025

出　　处：《中国血吸虫病防治杂志》2005年第4期255-259,共5页Chinese Journal of Schistosomiasis Control

基　　金：国家高技术研究发展计划(863计划)(No.2004AA215240;2004AA2Z3520);国家自然科学基金(No.30371262);上海市科委"十五"科技攻关重大计划(03DZ19231)~~

摘　　要：目的观察日本血吸虫(大陆株)硫氧还蛋白DNA疫苗(pcDNA3-SjcTrx)在小鼠诱导抗血吸虫感染的免疫保护作用。方法制备pcDNA3-SjcTrx重组质粒,将30只雌性C57BL/6小鼠随机分为3组,每组10只:pcDNA3-SjcTrx核酸疫苗免疫组、pcDNA3空质粒对照组和攻击感染对照组。核酸疫苗免疫组每只小鼠经股四头肌注射100μg核酸疫苗,共注射3次,间隔2周。空质粒对照组每只小鼠在相应时间经股四头肌注射100μgpcDNA3空质粒,感染对照组则不注射任何质粒。于末次免疫后3周,每只小鼠经腹部感染(30±1)条日本血吸虫尾蚴。小鼠于攻击感染后42天剖杀,门脉灌注收集成虫,计数成虫数和肝内虫卵数。分别在免疫前、攻击感染前和小鼠剖杀前采血并分离血清,用ELISA检测血清中特异性IgG抗体。另取6只雌性C57BL/6小鼠经股四头肌注射核酸疫苗,分别于注射后24、48小时和72小时取肌肉注射部位局部组织制备冰冻切片,用免疫酶染色试验(IEST)检测注射局部组织抗原的表达情况,以注射pcDNA3空质粒者为对照。结果IEST结果表明该DNA疫苗在小鼠肌肉组织内表达,ELISA检测表明DNA疫苗免疫小鼠后产生明显的抗体(IgG)免疫应答,并诱导出对攻击感染的45.7%的减虫率和41.4%的肝组织减卵率(P<0.05)。结论日本血吸虫(大陆株)硫氧还蛋白DNA疫苗具有较好的免疫原性,在小鼠诱导出明显的免疫保护作用,可作为疫苗候选分子作进一步的研究。Objective To prepare DNA vaccine of Schistosoma japonicum （Chinese strain）thioredoxin（pcDNA3-SjcTrx） and investigate the protective immunity efficacy of the DNA vaccine against schistosomiasis in mice. Methods The recombinant plasmid pcDNA3-SjcTrx was prepared and used for immunization in mice. Thirty C57TBL/6 female mice were divided into three groups, 10 mice in each group ： pcDNA3-SjcTrx group, pcDNA3 control group and challenging group. In pcDNA3-SicTrx group, each mouse was immunized intramuscularly with 100μg pcDNA3-SjcTrx at week 0,2 and 4. In pcDNA3 control group and challenging control group, each mouse was immunized with 100μg pcDNA3 plasmid DNA and saline in the same immunized schedule, respectively.Three weeks after final iniection, each mouse was challenged with （30±1） cercariae of S. japonicum（Chinese strain）. At 42 days after challenge, all mice were sacrificed and the worms were collected by perfusion. The number of recovered worms and eggs from hepatic tissue of mice were counted. The sera were collected from mice before immunization, before challenge and before killing. The anti-SjcTrx antibodies were detected with ELISA assay. Another six 6-week old C57BL/6 mice were immunized with pcDNA3-SjcTrx as above （i. m. ）, and the muscle tissues of injection area were treated for frozen section at the hours 24, 48, 72 after immunization. The antigen of pcDNA3-SjcTrx expressed in muscle of mice were detected by immunoenzymatic staining technique （IEST）. Results IEST result showed that pcDNA3-SjcTrx were expressed in the muscle tissue injection area. ELISA results showed that specific antibodies against SjcTrx developed in mice.Compared with challenging control group, the worm reduction rate and egg reduction rate in liver tissues of pcDNA3-SjcTrx group were 45.7% and 41.4%, respectively （P〈0.05）. Conclusion The DNA vaccine of Schistosoma japonicum （Chinese strain） thioredoxin had good immunogenicity and induced certain immunoprotective capacities against Schistosoma

关 键 词：日本血吸虫 硫氧还蛋白 DNA疫苗 小鼠 免疫 保护作用 

分 类 号：R383.24[医药卫生—医学寄生虫学]