摘要
目的 获得抗旋毛虫成虫的单克隆抗体 (单抗 )并初步研究其保护性免疫作用。方法 利用杂交瘤技术 ,用旋毛虫成虫抗原免疫的Balb/c小鼠脾细胞与小鼠骨髓瘤细胞SP2 / 0进行细胞融合。用免疫双扩散鉴定单抗亚类 ,免疫印迹鉴定单抗的特异性。小鼠尾静脉接种单抗进行被动免疫保护实验。结果 建立了 1株稳定分泌抗旋毛虫成虫单抗的细胞株。杂交瘤细胞培养上清液效价为1∶6 4 0 0 ,诱生腹水ELISA效价达 1∶2 0 4 80 0。经鉴定单克隆抗体类型为IgM ,可以识别旋毛虫成虫、肌幼虫及新生幼虫 ,相对分子质量约为 4 0 0 0 0蛋白 ,并且不与猪蛔虫、血吸虫、包虫、囊虫抗原发生交叉反应。单抗被动免疫后的减虫率为 5 5 .6 3%。结论 获得 1株具种特异性、保护性的单克隆抗体 。
Objective To prepare a monoclonal antibody(McAb) against Trichinella spiralis (T.s) adult and study its protective immunity. Methods BALB/c mice were immunized with soluble antigens of 3 day adult of T.s. Immnuized spleen cells of BALB/c mice were fused with myeloma cells sp2/0. The hybridoma culture supernatant was isotyped by the Ouchterlony double diffusion technique and the specificity of the McAb was determined by immunoblotting. The protective immunity of the McAb was detected after challenging the mice by the intravenous way. Results A McAb against T.s adult was obtained. The titer of culture fluid and ascetic fluid was 1∶6 400 and 1∶204 800, respectively. This McAb was identified as IgM. Western blotting results showed this McAb can be used to identify 40 000 protein of adult worm, muscle larvae and newborn larvae of T.s. No cross reactions with antigens of Ascaris suum Goeze, Taenia solium Linnaeus, Echinococcus granulosus and Schistosoma were oberserved. The number of muscle larvae was decreased by 55.63% when giving McAb to mice intravenously. Conclusions A species specific McAb against T.s adult was established and its protective immunity was identified. This McAb can be used as a powerful tool in screening Trichinella vaccine candidates.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2002年第5期297-300,共4页
Chinese Journal of Infectious Diseases
基金
美国中华医学基金会(CMB parasitology 98-674)
北京市教委科技发展基金资助项目
