Bowtell D D, Saint R B, Rickard M D, Mitchell G F
Parasitology. 1986 Dec;93 ( Pt 3):599-610. doi: 10.1017/s0031182000081300.
Previously we reported the isolation of several Escherichia coli clones expressing fragments of Taenia taeniaeformis antigens as beta-galactosidase fused proteins (Bowtell, Saint, Rickard & Mitchell, 1984). Here we describe the isolation of additional antigen-expressing clones from a larval cDNA library and the assignment of these clones to 7 antigen families. These were isolated with a polyspecific rabbit antiserum raised to the oncosphere. Since this serum was capable of reacting with a large number of antigens, it was important to develop techniques for rapidly determining the identity of the native T. taeniaeformis molecule corresponding to a cloned antigen gene. These included active immunization of rabbits with fused proteins and several techniques involving affinity purification on immobilized fused proteins. The reactivity of the antigen-positive clones with sera from humans infected with related parasites was also assessed. Finally, immunization of mice with several fused proteins failed to protect against subsequent infection, although antigens previously identified as candidate host-protective antigens (Bowtell, Mitchell, Anders, Lightowlers & Rickard, 1983) have yet to be identified in the expression library.
此前我们报道了从表达绦虫带状绦虫抗原片段的大肠杆菌克隆中分离出一些作为β-半乳糖苷酶融合蛋白的克隆(鲍特尔、圣、里卡德和米切尔,1984年)。在此,我们描述了从幼虫cDNA文库中分离出更多表达抗原的克隆,并将这些克隆归为7个抗原家族。这些克隆是用针对六钩蚴产生的多特异性兔抗血清分离得到的。由于该血清能够与大量抗原发生反应,因此开发快速确定与克隆抗原基因相对应的天然带状绦虫分子身份的技术很重要。这些技术包括用融合蛋白对兔子进行主动免疫以及几种涉及在固定化融合蛋白上进行亲和纯化的技术。还评估了抗原阳性克隆与感染相关寄生虫的人类血清的反应性。最后,用几种融合蛋白对小鼠进行免疫未能预防随后的感染,尽管在表达文库中尚未鉴定出先前被确定为候选宿主保护性抗原的抗原(鲍特尔、米切尔、安德斯、莱特沃勒斯和里卡德,1983年)。