Ito A, Bøgh H O, Lightowlers M W, Mitchell G F, Takami T, Kamiya M, Onitake K, Rickard M D
Department of Parasitology, Gifu University School of Medicine, Japan.
Mol Biochem Parasitol. 1991 Jan;44(1):43-9. doi: 10.1016/0166-6851(91)90219-v.
Primary screening of a cDNA expression library of Taenia taeniaeformis oncospheres in lambda gt11 bacteriophage was carried out using rabbit anti-T, taeniaeformis oncosphere serum affinity-purified from oncosphere pellets. From approximately 1.6 x 10(5) plaques, 21 single clones that were positive with the affinity-purified antibodies were isolated. Sibling analysis revealed that 17 clones out of the 21 could be assigned to five different antigen families. Only family 1 was strongly recognized by a serum prepared in a rabbit against a partially purified host-protective oncosphere antigen fraction. The fragments of lambda DNA were inserted into a pGEX plasmid vector that encodes glutathione S-transferase (GST) of Schistosoma japonicum. Clones designated TtO-18, -49.53 (family 1), 46 (family 2), 15 (family 3), 40 (family 4) and 66 (family 5) were established as subclones in pGEX-1 plasmid vectors which produced GST fusion proteins. All GST fusion proteins were soluble and recognized by anti-GST and anti-TtO sera. Three vaccination experiments with these fusion proteins using specific-pathogen-free Wistar rats revealed that all three fusion proteins of family 1 were exclusively effective against T. taeniaeformis oncosphere challenge with approximately 95% and 91% reductions in cystic metacestode and total metacestode recoveries, respectively. Rats vaccinated with fusion proteins of family 1 produced antibodies which reacted with a 21-kDa oncosphere antigen component which appeared to be a major oncosphere stage-specific antigen.
利用从六钩蚴沉淀中亲和纯化的兔抗豆状带绦虫六钩蚴血清，对λgt11噬菌体中的豆状带绦虫六钩蚴cDNA表达文库进行了初步筛选。从大约1.6×10⁵个噬菌斑中，分离出21个与亲和纯化抗体呈阳性反应的单克隆。同胞分析表明，21个克隆中有17个可归为5个不同的抗原家族。只有家族1被用兔制备的针对部分纯化的宿主保护性六钩蚴抗原组分的血清强烈识别。将λDNA片段插入编码日本血吸虫谷胱甘肽S-转移酶（GST）的pGEX质粒载体中。指定为TtO-18、-49.53（家族1）、46（家族2）、15（家族3）、40（家族4）和66（家族5）的克隆被构建为pGEX-1质粒载体中的亚克隆，这些载体产生GST融合蛋白。所有GST融合蛋白都是可溶的，并能被抗GST和抗TtO血清识别。使用无特定病原体的Wistar大鼠对这些融合蛋白进行的三次疫苗接种实验表明，家族1的所有三种融合蛋白对豆状带绦虫六钩蚴攻击均具有独特的效果，分别使囊尾蚴和总囊尾蚴回收率降低约95%和91%。用家族1的融合蛋白接种的大鼠产生了与一种21 kDa的六钩蚴抗原成分反应的抗体，该成分似乎是主要的六钩蚴阶段特异性抗原。
