日本血吸虫SjCa8基因的原核表达及其免疫学特征分析

目的表达和纯化日本血吸虫钙结合蛋白(SjCa8),研究其免疫学特征。方法重组质粒pET32a ( )-SjCa8经诱导表达后,获取纯化蛋白SjCa8,利用ELISA、Western-blot和动物保护性实验检测SjCa8的免疫学特征。结果成功获取了纯化蛋白SjCa8,该蛋白能免疫识别血吸虫感染小鼠血清和血吸虫病患者血清。SjCa8免疫Balb/c小鼠可获得较高滴度的特异性抗体,诱导宿主产生35．31%的减虫率和35．68%的减卵率。结论纯化蛋白SjCa8具有良好的免疫原性和免疫反应性,具有作为血吸虫病免疫诊断和疫苗候选分子的研究价值。

Prokaryotic expression of SjCa8 from Schistosoma japonicum and its immunological identification

Objective To express the gene encoding 8 kD calcium-binding protein from Schistosoma japonicum(SjCa8)and study its immunological characters.Methods SjCa8,expressed in pET32a( )-SjCa8/ BL21,was purified by His.Bind~(TM)purification kit.ELISA,Western-blot,flow cytometry and animal test were performed to assess the immunological characters of SjCa8.Results Purified SjCa8 could react with sera from mice infected by cercariae and sera from patients with infection.High titer antibody could be detected in the serum from mice vaccinated by SjCa8 and a significant reduction of adult worms(35.31%)and eggs(35.68%)was found in the animal test.Up-regulation of co-stimulatory molecules expression on dentritic cells(DC)by SjCa8 was evident. Conclusions SjCa8 shows a potential diagnostic value for schistosomiasis and might be a promising vaccine candidate against the plague.