旋毛虫丝氨酸蛋白酶抑制剂定位及免疫保护效果评估

LOCALIZATION AND IMMUNOPROTECTION OF TRICHINELLA SPIRALIS SERINE PROTEASE INHIBITORS

探讨旋毛虫丝氨酸蛋白酶抑制剂(Ts-serpin)的抗原性、定位及免疫保护性。将本实验室前期构建的旋毛虫丝氨酸蛋白酶抑制剂的原核表达载体进行大量表达纯化。利用不同感染时间的猪旋毛虫阳性血清,通过Western blot方法对纯化后的重组丝氨酸蛋白酶抑制剂(rTs-serpin)进行反应原性鉴定,并制备多克隆抗体,用间接免疫荧光法检测Ts-serpin在旋毛虫肌幼虫中的定位。随后将rTs-serpin免疫小鼠进行免疫保护效果评估。结果显示:rTs-serpin可被不同感染时间的猪旋毛虫阳性血清特异性识别,表明rTsserpin是高反应原性抗原;Western blot结果显示旋毛虫排泄分泌物和旋毛虫虫体粗提取物均存在Ts-serpin,间接免疫荧光显示Tsserpin定位在旋毛虫肌幼虫表皮中;免疫保护试验结果显示rTs-serpin免疫后的小鼠旋毛虫肌幼虫减虫率约为32.2%。综上所述,Tsserpin主要定位在旋毛虫肌幼虫表皮,rTs-serpin具有较强的抗原性,且对小鼠具有免疫保护作用。

The Trichinella spiralis serine protease inhibitor(Ts-serpin)was expressed in large amount in the prokaryoticb system to investigate its antigenicity,localization and immunoprotection.The recombinant Ts-serpin(rTs-serpin)was purified and used in Western blot to test positive sera collected at different infection stages,indicating that rTs-serpin is a highly reactive antigen.Ts-serpin existed in both excreted secretions and crude extracts of T.spiralis as demonstrated in Western blot,and localized in the epidermis of T.spiralis muscle larvae as visualized in indirect immunofluorescence assay.In addition,mice were immunized with rTs-serpin and challenged with T.spiralis muscle larvae.The immunized mice showed the worm reduction rate at 32.2%as compared with non-immunized controls.In summary,Ts-serpin mainly existed in the epidermis of T.spiralis muscle larvae and provided some protection in mice.