中国寄生虫学与寄生虫病杂志 ›› 2002, Vol. 20 ›› Issue (4): 3-204.

• 论著 • 上一篇    下一篇

日本血吸虫SjC21.7核酸疫苗诱导小鼠保护性免疫作用的研究

余传信;朱荫昌;殷旭仁;任建功;司进;许永良;沈林南
  

  1. 江苏省血吸虫病防治研究所 无锡214064
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2002-08-30 发布日期:2002-08-30

Protective Immunity Induced by the Nucleic Acid Vaccine of SjC 21.7 in Mice

YU Chuan-xin*;ZHU Yin-chang;YIN Xu-ren;REN Jian-gong;SI Jin;XU Yong-liang;SHEN Lin-nan
  

  1. Jiangsu Institute of Parasitic Diseases;Wuxi 214064
  • Received:1900-01-01 Revised:1900-01-01 Online:2002-08-30 Published:2002-08-30

摘要:   目的 研究日本血吸虫中国大陆株 2 1.7k Da膜蛋白分子 (Sj C2 1.7)核酸疫苗对 BAL B/ c小鼠的免疫保护性作用。 方法 采用 PCR方法扩增出特异性 Sj C2 1.7基因的开放阅读框序列 ,在其起始密码子处引入 Kozark序列。将目的基因片段亚克隆到真核表达质粒 pc DNA3.1中 ,构建重组真核表达质粒 Sj C2 1.7- pc DNA3.1。 4 8只 BAL B/ c小鼠分为对照组、实验组和加强组。对照组小鼠的股四头肌注射接种 pc DNA3.1,实验组同法注射重组质粒 Sj C2 1.7- pc D-NA3.1,加强组除注射重组质粒 Sj C2 1.7- pc DNA3.1外 ,同时注射重组质粒 P35 - pc DNA3.1及 P4 0 - pc DNA。每隔 2 wk免疫 1次 ,共免疫 3次。第 3次免疫后第 30天 ,每只鼠感染 4 5± 1条尾蚴 ,4 5 d后剖杀计数各组小鼠成虫数及肝卵数。通过 EL ISA和免疫组化分析观察小鼠免疫学特征的变化。 结果 免疫组化分析显示 ,实验组小鼠股四头肌局部组织有特异性抗原蛋白表达。EL ISA分析表明 ,免疫后实验组和加强组有部分小鼠出现特异性 Ig G抗体。与对照组比较 ,实验组减虫率及减卵率分别为 2 9.9%及 13.8% ,加强组分别为 31.9%及 2 8.0 %。加强组减卵率显著高于实验组(P<0 .0 5 )。 结论  Sj C2 1.7核酸疫苗能诱导 BAL B/ c小鼠产生一定水平的抗血吸虫感染

关键词: 日本血吸虫, 21.7kDa蛋白分子, 核酸疫苗, 免疫保护作用, IL-12

Abstract:  Objective To observe the protective immunity induced by the nucleic acid vaccine of 21.7 kDa membrane protein molecule of Schistosoma japonicum Chinese mainland strain (SjC 21.7) in BALB/c mice. . Methods. A pair of primers (P1 and P2) was synthesized according to the DNA sequence of the SjC21.7. The ORF sequence of SjC21.7 was amplified by PCR, and the Kozark sequence was added to the position of initiator. The gene fragment was inserted into the eukaryotic expression plasmid pcDNA3.1 to form the recombinant plasmid SjC21.7-pcDNA3.1. Forty-eight BALB/c mice were divided into three groups: control, test and boost. Each mouse was injected in quadriceps femoris with plasmid pcDNA3.1 (control) or recombinant plasmid SjC21.7-pcDNA3.1 (test, boost); for the boost group, with additional P35-pcDNA3.1 and P40-pcDNA3.1. All mice were immunized three times with an interval of 2 weeks, challenged each with 45 cercariae of S.japonicum at the 30th day after final immunization. At day 45 after challenge,all mice were sacrificed, the numbers of worms and hepatic eggs were counted. Antibody level in the sera of mice before and two weeks after immunization was determined with ELISA. The expression of the target gene in quadriceps femoris was observed with immunohistochemistry. . Results . The immunohistochemistry analysis showed that there were specific antigens expressed in the local tissue of the test group mice. There was specific IgG in the serum of partial mice in test and boost groups. Compared with the control group, the worm reduction rate was 29.9% and its egg reduction rate 13.8% in the test group; 31.9% and 28.0% respectively in the boost group. The egg reduction rate in the boost group was higher than that of the test group (P<0.05). . Conclusion . The SjC21.7 nucleic acid vaccine could induce partial protective immunity against Schistosoma japonicum in BALB/c mice.

Key words: Schistosoma japonicum, 21.7 kDa membrane protein, nucleic acid vaccine, protective immunity, IL-12