论文部分内容阅读
目的探讨重组日本血吸虫四跨膜蛋白第二亲水基团融合蛋白(GST-TSP2HD)抗血吸虫感染免疫保护效果。方法采用Glutathione sepharose 4B胶亲和层析法大量制备GST-TSP2HD蛋白。实验组以GST-TSP2HD融合蛋白的福氏佐剂抗原免疫C57BL/6J小鼠,同时设置佐剂对照组及GST蛋白免疫对照组,每2周加强免疫1次。加强免疫2次后每只小鼠经腹部皮肤感染(45±2)条血吸虫尾蚴。感染42 d后剖杀小鼠,经生理盐水静脉灌注收集成虫,计数减虫率,用KOH溶液消化肝组织收集虫卵,计算减卵率。组织切片观察小鼠肝组织病理变化,通过酶联免疫吸附试验检测小鼠血清中的抗体水平。结果与佐剂对照组相比,GST-TSP2HD融合蛋白免疫组的减虫率为27.10%,减卵率为32.30%;与GST蛋白免疫组相比,GST-TSP2HD融合蛋白免疫组可获得39.57%的减虫率,43.53%的减卵率。GST-TSP2HD融合蛋白免疫组肝组织中的虫卵肉芽肿数量明显减少,单个虫卵肉芽肿平均直径比佐剂对照组、GST蛋白免疫组分别下降了27.08%(P<0.05)和40.53%(P<0.01),差异均有统计学意义。GST-TSP2HD能诱导高水平抗TSP2HD蛋白的特异性抗体IgG,其中IgG1、IgG2b和IgG2c亚类可能在小鼠抗血吸虫感染保护性免疫中发挥重要作用。结论日本血吸虫四跨膜蛋白是一个有效的日本血吸虫病疫苗候选分子。
Objective To investigate the protective effect of GST-TSP2HD against Schistosoma japonicum infection by recombinant Schistosoma japonicum tetra-transmembrane protein. Methods GST-TSP2HD protein was prepared in large quantities by Glutathione sepharose 4B gel electrophoresis. In the experimental group, C57BL / 6J mice were immunized with Freund’s adjuvant antigen of GST-TSP2HD fusion protein. Adjuvant control group and GST protein immunization control group were also set up, and the immunization was boosted once every two weeks. After two boosts, each mouse was infected (45 ± 2) with cercariae by abdominal skin. Mice were sacrificed after 42 days of infection and adult worms were harvested by intravenous infusion of saline. The worm reduction rate was counted. The eggs were harvested by digestion of liver tissue with KOH solution to calculate the oviposition rate. Tissue sections were used to observe the pathological changes of liver tissue in mice. The antibody levels in serum of mice were detected by enzyme linked immunosorbent assay (ELISA). Results Compared with the adjuvant control group, the GST-TSP2HD fusion protein immunization group had an anti-worm rate of 27.10% and an egg reduction rate of 32.30%. Compared with the GST protein-immunized group, GST-TSP2HD fusion protein immunization group achieved 39.57% Of the worm reduction rate, 43.53% of the egg reduction rate. Compared with the adjuvant control group and the GST protein immunization group, the average diameter of single egg granuloma decreased by 27.08% (P <0.05) and 40.53% (P <0.05), respectively, compared with the GST-TSP2HD fusion protein immunization group <0.01), the differences were statistically significant. GST-TSP2HD can induce high-level anti-TSP2HD protein-specific IgG, IgG1, IgG2b and IgG2c subclasses may play an important role in protective immunity against infection by mouse anti-schistosome. Conclusion The four-transmembrane protein of Schistosoma japonicum is an effective vaccine candidate for schistosomiasis japonica.