日本血吸虫HGPRT重组抗原与不同佐剂联合应用对小鼠免疫保护作用的研究  

Protective immunity induced by recombinant Schistosoma hypoxanthine-guanine phosphoribosyltransferase with different adjuvants in mice

作　　者：胡媛[1] 沈玉娟[1] 曹建平[1] 徐馀信[1] 卢潍媛[1] 周何军[1] 张璟[1] 李小红[1] 权红[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：《江苏大学学报（医学版）》2010年第2期93-97,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(30771880;30872212);国家高技术研究发展计划基金资助项目(国家863计划)(2006AA02Z444);国家传染病重大科技专项基金资助项目(2009ZX10004-201;2008ZX10004-002);上海市科委科技攻关重大计划项目(064319026)

摘　　要：目的:观察日本血吸虫(大陆株)次黄嘌呤鸟嘌呤磷酸核糖转移酶(hypoxanthine-guanine phosphoribosyl-transferase,HGPRT)重组抗原(reSjc HGPRT)与ISA206或弗氏佐剂联合免疫对小鼠诱导抗日本血吸虫感染的保护作用。方法:雌性C57BL/6小鼠随机分为5组:reSjc HGPRT加ISA206佐剂免疫组、reSjc HGPRT加弗氏佐剂组、ISA206佐剂对照组、弗氏佐剂对照组和感染对照组。20μg重组抗原和ISA206或弗氏佐剂乳化后小鼠项背部多点皮下注射,共免疫3次,每次间隔2周。佐剂对照组小鼠仅注射ISA206或弗氏佐剂和生理盐水,感染对照组不注射任何重组抗原或生理盐水。于末次免疫后3周,每只小鼠经腹部皮肤感染(30±1)条日本血吸虫尾蚴,6周后剖杀小鼠,门脉灌注收集成虫,计数成虫数、雌雄合抱数和小鼠肝组织虫卵数。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检测血清中特异性IgG抗体。结果:重组抗原加ISA206佐剂或弗氏佐剂免疫组均诱导小鼠产生特异性IgG抗体应答,与感染对照组和弗氏佐剂对照组相比,差异有统计学意义(P<0.05);诱导小鼠产生的减虫率、减雌雄合抱率和肝组织减卵率分别为53.7%、59.3%、44.9%和43.3%、44.1%、33.0%,与感染对照组和2种佐剂对照组相比均有统计学意义(P<0.05)。结论:reSjcHGPRT与ISA206或弗氏佐剂联合免疫,可诱导小鼠产生抗血吸虫感染保护作用。Objective：To investigate the protective immunity of recombinant hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum（reSjc HGPRT）plus Montanide ISA 206 or Freund＇s adjuvants in mice.Methods：Female C57BL/6 mice were randomly divided into five groups,reSjc HGPRT plus Montanide ISA 206 adjuvant,reSjc HGPRT plus Freund＇s adjuvant,two adjuvant control groups and challenged control group.In Montanide ISA 206 adjuvant or two groups of reSjc HGPRT plus Freund＇s adjuvant,each mouse was immunized subcutaneously with 20 μg reSjc HGPRT emulsified in Montanide ISA 206 or FCA/FIA,respectively,while in two adjuvant control group,each mouse was injected subcutaneously with sterile normal saline emulsified in FCA/FIA or Montanide ISA 206 respectively.For challenged control group,mice were not treated with recombinant antigen or adjuvant.All mice were vaccinated for three times in an interval of 2 weeks.Two weeks after final immunization,each mouse was challenged with（30±1） cercariae of S.japonicum.At the sixth week after challenged,all mice were sacrificed and the number of worms,pairs of worm and eggs in the livers were counted.The sera collected respectively from mice before immunized,challenged and killed were identified by ELISA assay to detect specific anti-SjcHGPRT IgG antibody.Results：The levels of specific IgG antibody in two immunized groups with reSjc HGPRT was higher than that of challenged control group.Also,compared with the challenged control,the worm reduction rates,worm in pairs rates and egg reduction rates in mice immunized with reSjc HGPRT plus Montanide ISA 206 adjuvant and plus Freund＇s adjuvant were 53.7%,59.3%,44.9% and 43.3%,44.1%,33.0%,respectively（P0.05）.Conclusion：A better immune protection could be obtained in mice immunized with recombinant antigen plus plus Montanide ISA 206 or Freund＇s adjuvants against S.japonicum.

关 键 词：日本血吸虫 次黄嘌呤鸟嘌呤磷酸核糖转移酶重组抗原 佐剂 免疫保护性 

分 类 号：R383.24[医药卫生—医学寄生虫学]