日本血吸虫SIEA66-68kDa抗原的分离、纯化及免疫保护性研究

Isolation and purification of the 66-68 kDa soluble immature egg antigen Schistosoma japonicum and its protective immunity

目的研究日本血吸虫未成熟虫卵可溶性抗原66-68kDa(SIEA66-68kDa)的动物免疫保护力,并对其作为天然分子疫苗的可行性进行评估。方法采用电泳切胶、电洗脱、超滤离心等技术,分离纯化SIEA66-68kDa天然分子抗原,免疫小鼠,待其产生抗体后进行尾蚴攻击感染(40尾/只),计算减虫率和减卵率。结果 SIEA66-68kDa天然蛋白质分子能刺激机体产生抗日本血吸虫的作用,其减虫率为41.70%,每克肝脏减卵率为51.23%,雌虫子宫减卵率为29.30%,每克大肠减卵率为59.26%,每克粪便减卵率为45.17%,与对照组相比差异具有统计学意义(P<0.05)。结论 SIEA66-68kDa天然分子抗原具有刺激小鼠产生较好的抗日本血吸虫感染的免疫保护作用,可作为抗日本血吸虫感染候选分子疫苗的靶标。

To investigate the immunoprotection of 66-68 kDa soluble antigen of immature egg of Schistosoma japonicum（SIEA66-68kDa） and to evaluate its feasibility of being used as a molecular vaccine antigen,the SIEA66-68kDa was prepared through SDS-PAGE electrophoresis,protein band resection,electro-elution and ultrafiltration,and the isolated product of protein was used as antigen to immunize Kunming strain of mice.After 3 courses of immunization,mice were challenged with cercariae of S.japonicum,and the reduction rates of worms and eggs were compared with those of the control group with inoculation of PBS.It was found that the mono-specific antibody response against SIEA66-68kDa protein could be induced in mice immunized with this protein.The SIEA66-68kDa could induce significant reduction in adult worms（41.70%）.The rates of egg reduction in the SIEA66-68kDa group reach 51.23%,59.26% and 45.17% in the liver,intestine tissue and feces,respectively,with significant difference when compared with the control group.It is concluded that the 66-68 kDa immature egg antigen of S.japonicum could protect the Kunming strain of mice from the infection of this parasite,and it could be regarded as the target antigen for the preparation of vaccine against S.japonicum infection.