中国寄生虫学与寄生虫病杂志
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中国寄生虫学与寄生虫病杂志 2006, Vol. 24 Issue (6) :440    DOI:
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日本血吸虫Sj23DNA疫苗的增效研究
朱艳红1;韩庆霞2;任伟2;牛安欧2;李柳哲2
华中科技大学同济医学院 1 医学生物学系; 2 病原生物学系, 武汉 430030
Studies on the Protective Effect of the Mutant of Sj23 DNAVaccine against Schistosomiasis
ZHU Yan-hong1;HAN Qing-xia2;REN Wei2;NIU An-ou2;LI Liu-zhe2
Departments of Biology1 and Parasitology2, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

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摘要 目的 探讨缺失肿瘤转移有关基因ME491同源序列的日本血吸虫Mr23 000膜蛋白基因(Sj23DNA)突变体疫苗的免疫效果。 方法 将Sj23DNA中与人黑素瘤细胞膜上的ME491同源序列进行基因缺失突变,再将突变基因转染真核细胞人胚肾HEK293细胞,通过间接荧光抗体试验(IFAT)检测其表达情况。将pcDNA3-Sj23突变体经股四头肌注射免疫小鼠(100 μg/只),免疫后4周取注射部位肌肉作冰冻切片,IFAT检测目的基因的表达。40只BALB/c小鼠随机分成4组(每组10只),分别经股四头肌注射pcDNA3-Sj23突变体、pcDNA3-Sj23、空白质粒pcDNA3(均100 μg/只)和生理盐水(30 μl/只)免疫1次。免疫后4周用尾蚴攻击感染(40±2条/只)。第42天剖杀,肠系膜静脉灌注法及挤压法计数成虫,取肝脏计算每克肝组织虫卵数(EPG)。以减虫率和减卵率评价其免疫保护力。 结果 pcDNA3-Sj23突变体疫苗可在真核细胞HEK293中瞬时表达。pcDNA3-Sj23突变体在小鼠注射部位骨骼肌细胞上出现较强荧光,空白对照组未见特异性荧光。pcDNA3-Sj23突变体获得40.3%减虫率和42.8%减卵率,pcDNA3-Sj23获得33.1%减虫率和28.9%减卵率。两组差异均有统计学意义(P值均<0.05)。 结论 与pcDNA3-Sj23相比,缺失ME491同源序列的cDNA3-Sj23突变体疫苗能诱导BALB/c小鼠产生更好的免疫保护效果。
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朱艳红
韩庆霞
任伟
牛安欧
李柳哲
关键词日本血吸虫   膜蛋白基因   肿瘤转移有关基因   ME491   免疫保护     
AbstractObjective To investigate the protective immunity of the vaccine against schistosomiasis, a mutant of Mr 23 000 membrane protein DNA (Sj23DNA) without the homologous sequence of ME491. Methods The mutant of Sj23 DNA with no homologous sequence of ME491 on the cell membrane of human melanoma was obtained by overlap PCR. The mutant was transfected into human embryonic kidney cells of the line HEK293. Indirect fluorescent antibody test (IFAT) was used to detect the expressed protein. Expression of the mutant of Sj23DNA in muscular cells of mice was conducted through vaccinating the mouse with 100 μg purified plasmids by injecting them into the quadriceps muscle of thigh. Four weeks after the immunization, the quadriceps muscles were taken and cryostat sections were prepared for detecting the expression by IFAT. Forty BALB/c mice were randomly divided into four groups and injected with the mutant of pcDNA3-Sj23 plasmid DNA, pcDNA3-Sj23 plasmid DNA, pcDNA3 blank plasmid (100 μg per mouse) and sterile saline (30 μl per mouse) respectively. Four weeks after the immunization, mice were challenged with cercariae (40±2 cercariae per mouse) by abdominal skin penetration. Mice were then killed 6 weeks later, perfusion and squash methods were carried out to collect the adult worms and the number of eggs per gram of liver tissue was calculated. Worm and egg reduction rates were used to evaluate the protective immunity. Results Specific fluorescence was demonstrated in muscular cells of mice vaccinated with the mutant of pcDNA3-Sj23. The worm reduction rate and egg reduction rate were 40.3% and 42.8% respectively in the mutant of pcDNA3-Sj23 group, which were higher than those in the pcDNA3-Sj23 plasmid group (33.1% and 28.9% respectively). The difference between these two groups was significant(P<0.05). Conclusion The modified Sj23DNA without the homologous sequence of ME491 induces higher protection against Schistosoma japonicum infection in mice than that of Sj23DNA.
Keywords Schistosoma japonicum   Sj23 DNA vaccine   ME491   Immunoprotection     
Received 1900-01-01; published 2006-12-30
Corresponding Authors: 牛安欧   
引用本文:   
朱艳红;韩庆霞;任伟;牛安欧;李柳哲.日本血吸虫Sj23DNA疫苗的增效研究[J]  中国寄生虫学与寄生虫病杂志, 2006,V24(6): 440
ZHUYan-hong;HANQing-xia;RENWei;NIUAn-ou;LILiu-zhe.Studies on the Protective Effect of the Mutant of Sj23 DNAVaccine against Schistosomiasis[J]  , 2006,V24(6):440
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