日本血吸虫幼虫巨大致死基因片段的克隆、表达及免疫效果分析  

Cloning and expressing the lethal giant larvae (LGL) gene fragment of Schistosoma japonnicum and evaluating the immunoprotective effect of truncated LGL

作　　者：石艳丽[1,2] 刘萍萍[2] 杨云霞[2] 刘金明[2] 林矫矫[2] 宋铭忻[1] 金亚美[2] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国预防兽医学报》2011年第7期557-561,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30800819);上海浦江人才(08PJ1411800)

摘　　要：为研究日本血吸虫(Schistosoma japonnicum)幼虫巨大致死基因(Lethal giant larvae)的生物学功能,本研究应用荧光定量RT-PCR分析SjLgL基因在S.japonnicum不同发育阶段的表达情况,构建重组表达质粒pET-SjLgL进行诱导表达,对重组蛋白进行western blot鉴定;重组蛋白刺激免疫动物进行免疫保护试验,检测IL-4、IL-10、TNF-α和IFN-γ的变化。结果表明,SjLgL基因在S.japonnicum不同发育时期均有表达,成虫期略高于童虫,雄虫高于雌虫。Western blot鉴定结果显示,该重组蛋白具有良好的抗原性。免疫保护试验结果表明,免疫小鼠分别获得了22.34%的减虫率和55.49%的肝脏减卵率。血清学检测结果显示重组蛋白免疫小鼠后产生了特异性IgG抗体。细胞因子检测结果表明,重组蛋白刺激免疫动物主要诱发产生Th1型的免疫应答。本实验结果表明该重组蛋白在小鼠体内可诱导产生部分免疫保护效果。To study immunoprotective effect of Lethal giant larvae （LGL） gene encoded protein of Schistosoma japonnicum （Sj）, we cloned and expressed the cDNA fragment of SjLGL in E.coli. and the immunoprotective activity of the truncated recombinant SjLGL was evaluated in immunized mice. The result showed that the SjLGL was expressed at several different devolopment stages of S. japonicum, and the transcription level of male adult worm was higher than the female. It was also indicated that rSjLGL was able to induce specific antibodies and the cytokine profile analysis demonstrated the induction of a Th1 immune profile in response to immunization with the truncated recombinant protein. Challenge of the mice immunized with rSjLGL demonstrated a 22.34% and 55.49% reduction of the mean worm burden and liver egg burden, respectively.

关 键 词：日本血吸虫 SjLGL 荧光实时定量PCR 免疫保护效果 

分 类 号：S852.72[农业科学—基础兽医学]