Torres J M, Sánchez C, Suñé C, Smerdou C, Prevec L, Graham F, Enjuanes L
Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, Madrid, Spain.
Virology. 1995 Nov 10;213(2):503-16. doi: 10.1006/viro.1995.0023.
Ten recombinant adenoviruses expressing either fragments of 1135, 1587, or 3329 nt or the full-length spike gene of transmissible gastroenteritis coronavirus (TGEV) have been constructed. These recombinants produce S polypeptides with apparent molecular masses of 68, 86, 135, and 200 kDa, respectively. Expression of the recombinant antigen driven by Ad5 promoters was inhibited by the insertion of an exogenous SV-40 promoter. Most of the recombinant antigens remain intracytoplasmic in infected cells. All the recombinant-directed expression products contain functional antigenic sites C and B (Gebauer et al., 1991, Virology 183, 225-238). The recombinant antigen of 135 kDa and that of 200 kDa, which represents the whole spike protein, also contain antigenic sites D and A, which have previously been shown to be the major inducers of TGEV-neutralizing antibodies. Interestingly, here we show that recombinant S protein fragments expressing only sites C and B also induced TGEV-neutralizing antibodies. The chimeric Ad5-TGEV recombinants elicited lactogenic immunity in hamsters, including the production of TGEV-neutralizing antibodies. The antisera induced in swine by the Ad5 recombinants expressing the amino-terminal 26% of the spike protein (containing sites C and B) or the full-length spike protein, when mixed with a lethal dose of virus prior to administration to susceptible piglets, delayed or completely prevented the induction of symptoms of disease, respectively.
已构建出十种重组腺病毒,它们分别表达传染性胃肠炎冠状病毒(TGEV)1135、1587或3329个核苷酸的片段或全长刺突基因。这些重组体分别产生表观分子量为68、86、135和200 kDa的S多肽。由Ad5启动子驱动的重组抗原的表达受到外源SV - 40启动子插入的抑制。大多数重组抗原在感染细胞中保留在细胞质内。所有重组导向的表达产物都含有功能性抗原位点C和B(Gebauer等人,1991年,《病毒学》183卷,225 - 238页)。135 kDa的重组抗原和代表整个刺突蛋白的200 kDa的重组抗原还含有抗原位点D和A,先前已证明它们是TGEV中和抗体的主要诱导剂。有趣的是,我们在此表明仅表达位点C和B的重组S蛋白片段也能诱导TGEV中和抗体。嵌合的Ad5 - TGEV重组体在仓鼠中引发了泌乳免疫,包括产生TGEV中和抗体。当用表达刺突蛋白氨基末端26%(含位点C和B)的Ad5重组体或全长刺突蛋白在给易感仔猪给药前与致死剂量的病毒混合时,所诱导的猪抗血清分别延迟或完全预防了疾病症状的出现。