日本血吸虫硫氧还蛋白谷胱甘肽还原酶免疫功能的研究

发布时间：2018-07-07 10:37

本文选题：日本血吸虫 + 硫氧还蛋白谷胱甘肽还原酶　；参考：《中国血吸虫病防治杂志》2015年04期

【摘要】：目的研究日本血吸虫硫氧还蛋白谷胱甘肽还原酶(Thioredoxin Glutathione Reductase of Schistosoma japonicum,SjTGR)的免疫原性及抗日本血吸虫感染的免疫保护性作用。方法 75只小鼠随机分为5组:空白组、PBS组、CpG2免疫组、TGR免疫组和TGR+CpG2联合免疫组,免疫3次。首次免疫前及第3次免疫后2周经尾静脉采血,采用酶联免疫法分别检测血清中抗SjTGR蛋白IgG、IgG1和IgG2a的抗体水平。第3次免疫后2周,每只小鼠经腹部感染日本血吸虫尾蚴40±2条,42 d后剖杀,收集成虫和虫卵,计算减虫率和减卵率;制备各组小鼠单个脾淋巴细胞,采用流式细胞术检测脾细胞表面标志物CD44high、CD4+CD44high或CD8+CD44high水平;用SjTGR刺激免疫小鼠脾细胞72 h,采用酶联免疫法检测脾细胞的IL-2、IL-4、IL-10和IFN-γ的水平。结果第3次免疫后2周TGR与CpG2+TGR免疫组小鼠血清抗SjTGR抗体的IgG滴度均达1:200 000以上,IgG2a与IgG1的比值(IgG2a/IgG1)随时间的延长缓慢增高。TGR免疫组和TGR+CpG2联合免疫组细胞上清中的IFN-γ和IL-2水平与空白组、PBS组、CpG2免疫组相比明显增高(P0.05)。TGR免疫组与TGR+CpG2免疫组小鼠脾细胞的CD44high、CD8+CD44high及CD4+CD44high与空白组和PBS组相比细胞比值增加(P0.05)。TGR免疫组和CpG2+TGR免疫组的减虫率分别为9.4%,10.5%,减卵率分别为9.2%和32.8%。结论 SjTGR具有较强的免疫原性,可诱导小鼠免疫反应向Th1型极化,但不足以产生抗日本血吸虫感染的保护效应。CpG2 ODN可能是一种有效的Th1型免疫佐剂。
[Abstract]:Objective to study the immunogenicity of thioredoxin Glutathione Reductase of Schistosoma japonicum SjTGR and its protective effect on Schistosoma japonicum infection. Methods Seventy-five mice were randomly divided into five groups: the control group (PBS) was immunized with CpG2 and the control group was immunized with TGR and TGR CpG2. Blood samples were collected from caudal vein before the first immunization and 2 weeks after the third immunization. The levels of anti-SjTGR IgG _ 1 and IgG _ 2a in serum were detected by enzyme-linked immunosorbent assay (Elisa). Two weeks after the third immunization, each mouse was infected with Schistosoma japonicum cercariae 40 卤2 cercariae by abdominal cavity for 42 days. The adult worm and eggs were collected, the worm reduction rate and egg reduction rate were calculated, and a single spleen lymphocyte was prepared in each group. The levels of CD44high CD4 CD44high or CD8 CD44high were detected by flow cytometry, and the levels of IL-2IL-4IL-10 and IFN- 纬 were detected by enzyme-linked immunosorbent assay (Elisa) after SjTGR stimulated spleen cells in mice for 72 h. Results the IgG titers of anti-SjTGR antibody in serum of TGR and CpG2 TGR immunized mice reached 1: 200 000 or more (IgG2a / IgG1) slowly increased with the time prolonging. The supernatant of TGR immunized group and TGR CpG2 combined immunized group was increased slowly. The levels of IFN- 纬 and IL-2 were significantly higher than those of the control group (P0.05). The percentage of CD44highpCD8 CD44high and CD4 / CD44high in spleen cells of mice immunized with TGR and TGR CpG2 were significantly higher than those in the control group and PBS group (P0.05). The worm abatement in TGR immunized group and CpG2 TGR immunized group was higher than that in PBS group (P0.05). The rate of egg reduction was 9.2% and 32.8g, respectively. Conclusion SjTGR has strong immunogenicity and can induce the immune response of mice to Th1 type polarization, but it is not sufficient to produce protective effect against Schistosoma japonicum infection. CpG2 ODN may be an effective Th1 type immune adjuvant.
【作者单位】：江苏省血吸虫病防治研究所、卫生部寄生虫病预防与控制技术重点实验室、江苏省寄生虫病分子生物学重点实验室;
【基金】：国家自然科学基金(81201316;30972581) 江苏省自然科学基金(BK2012544) 国家传染病重大专项(2012ZX1004-220) 江苏省卫生厅血地寄科技项目(X201429;X201402)
【分类号】：R383.24

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