Dalton J P, Tom T D, Strand M
Proc Natl Acad Sci U S A. 1987 Jun;84(12):4268-72. doi: 10.1073/pnas.84.12.4268.
Spleen cells of mice vaccinated with radiation-attenuated Schistosoma mansoni cercariae were used to produce monoclonal antibodies directed against newly transformed schistosomular surface antigens. One of these monoclonal antibodies recognized a polypeptide of 18 kDa. This glycoprotein was purified by monoclonal antibody immunoaffinity chromatography and a polyclonal antiserum was prepared against it. Immunofluorescence assays showed that the polyclonal antiserum bound to the surface of newly transformed schistosomula and lung-stage organisms but not to the surface of liver-stage and adult worms. Using this polyclonal antiserum we isolated recombinant clones from an adult worm cDNA expression library constructed in lambda gt11. Clone 654.2 contained an insert of 0.52 kilobase and hybridized to a 1.2-kilobase mRNA species from adult worms. Most importantly, clone 654.2 produced a fusion protein of 125 kDa that was reactive with sera of vaccinated mice that are capable of transferring resistance. This result encourages future vaccination trials with the fusion protein.
用经辐射减毒的曼氏血吸虫尾蚴免疫的小鼠脾细胞来制备针对新转化的血吸虫表面抗原的单克隆抗体。其中一种单克隆抗体识别出一条18 kDa的多肽。通过单克隆抗体免疫亲和层析法纯化该糖蛋白,并制备了针对它的多克隆抗血清。免疫荧光试验表明,该多克隆抗血清与新转化的血吸虫幼虫和肺期虫体的表面结合,但不与肝期虫体和成虫的表面结合。利用这种多克隆抗血清,我们从构建于λgt11的成虫cDNA表达文库中分离出重组克隆。克隆654.2含有一个0.52千碱基的插入片段,并与来自成虫的1.2千碱基mRNA杂交。最重要的是,克隆654.2产生了一种125 kDa的融合蛋白,该蛋白与能够传递抗性的免疫小鼠血清发生反应。这一结果鼓励了未来对该融合蛋白进行疫苗试验。