中国寄生虫学与寄生虫病杂志 ›› 2006, Vol. 24 ›› Issue (3): 4-178.

• 论著 • 上一篇    下一篇

日本血吸虫重组硫氧还蛋白小鼠保护性免疫研究

韩海勃;曹建平;刘述先;徐裕信;沈玉娟;李小红;卢潍媛;刘海鹏;汤林华   

  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, 上海 20025
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-06-30 发布日期:2006-06-30
  • 通讯作者: 曹建平

Protective Immunity Induced by Recombinant Schistosomajaponicum Thioredoxin in Mice

HAN Hai-bo;CAO Jian-ping;LIU Shu-xian;XU Yu-xin;SHEN Yu-juan;LI Xiao-hong;LU Wei-yuan;LIU Hai-peng;TANG Lin-hua   

  1. National Institute of Parasitic Disease, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, MOH, Shanghai 200025, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-06-30 Published:2006-06-30
  • Contact: CAO Jian-ping

摘要: 目的 观察日本血吸虫(大陆株)硫氧还蛋白重组抗原在小鼠诱导抗血吸虫感染的免疫保护作用。 方法 30只雌性C57BL/6小鼠随机分为3组,reSjcTrx免疫组:reSjcTrx(15mg/鼠)和ISA720佐剂乳化后采用小鼠相背部多点皮下注射,共免疫3次,间隔2周;ISA720佐剂对照组:小鼠仅注射ISA720佐剂和生理盐水;感染对照组不作任何处理。于末次免疫后3周,各组小鼠经腹部皮肤感染(30±1)条日本血吸虫尾蚴,感染后6周剖杀,门静脉灌注法收集成虫,计成虫数和每克肝组织虫卵数。在免疫前、攻击感染前和小鼠剖杀前分别采血并分离血清,用ELISA检测重组抗原特异性IgG抗体。并对reSjcTrx 进行十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹(Western blotting)分析。 结果 ELISA检测表明,reSjcTrx免疫组小鼠产生特异性IgG抗体应答,并诱导小鼠产生对攻击感染的减虫率和肝组织减卵率分别为22.8%和29.5%,与ISA720佐剂对照组和感染对照组相比,差异均有统计学意义(P﹤0.05)。SDS-PAGE和Western blotting的结果表明,该重组抗原相对分子质量(Mr)约14 000(含载体的6个氨基酸),可被日本血吸虫感染兔血清和reSjcTrx免疫小鼠血清所识别。 结论 reSjcTrx免疫小鼠可产生一定的抗血吸虫感染的保护作用。

关键词: 日本血吸虫, 硫氧还蛋白, 重组蛋白, 免疫保护力

Abstract: Objective To investigate the protective immunity of the recombinant thioredoxin of Schistosoma japonicum(reSjcTrx)in mice. Methods Thirty 6-week old female C57BL/6 mice were randomly divided into 3 groups with 10 each: reSjcTrx with Montanide ISA720 adjuvant, adjuvant control, and infection control. Mice were vaccinated subcutaneously at week 0, 2, 4 with reSjcTrx emulsified in Montanide ISA720 adjuvant. The mice in adjuvant group was injected three times with Montanide ISA720 and saline only. Mice in infection control group were given no injection. Three weeks after final injection, each mouse was challenged with 30±1 cercariae of S. japonicum (Chinese strain). At the week six after challenge, all mice were sacrificed and perfused. The number of recovered worms and eggs from liver tissue of mice were counted. Sera were collected from mice before immunization, before challenge and before killing. The anti-SjcTrx antibodies in sera were detected with ELISA. Results ELISA showed a high level of specific IgG antibodies in mice immunized with the reSjcTrx. The worm reduction rate and egg reduction rate of reSjcTrx immunization group were 22.8% and 29.5% respectively, significantly higher than those of the control groups (P﹤0.05). SDS-PAGE and Western blotting revealed that the molecular weight of expressed protein was around Mr 14 000 and could be recognized by sera from rabbit infected with S.japonicum and from mice immunized with reSjcTrx. Conclusion The reSjcTrx induces certain protective immunity against schistosomiasis japonica in mice.

Key words: Schistosoma japonicum, Thioredoxin, Recombinant protein, Immunity