An adenovirus recombinant expressing the spike glycoprotein of porcine respiratory coronavirus is immunogenic in swine
An adenovirus recombinant expressing the spike glycoprotein of porcine respiratory coronavirus is immunogenic in swine
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DOI：
10.1099/0022-1317-77-2-309
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发表时间：
1996-02-01
影响因子：
3.8
通讯作者：
Pensaert, M
中科院分区：
文献类型：
--
作者：
Callebaut, P;Enjuanes, L;Pensaert, M
将猪呼吸道冠状病毒（PRCV）S基因全长插入人5型腺病毒基因组早期转录区3启动子下游。重组病毒在猪睾丸ST细胞系的培养物中复制，并指导S抗原的合成，最大产量约为26 μ g/10（6）个细胞。除了在感染的晚期，当少量（3.5 μ g/10（6）个细胞）被释放时，抗原是细胞相关的。细胞相关抗原由分子量为160 kDa和175 kDa的多肽组成，分别与PRCV的真实前体S'和成熟S蛋白共迁移。胞外重组抗原对应于175 kDa的成熟蛋白。一些重组S蛋白暴露在细胞表面，并被中和介导的抗S单克隆抗体识别。仔猪，经口鼻接种与重组腺病毒载体PRCV中和血清抗体和PRCV攻击的部分保护，证明了活腺病毒作为疫苗载体的潜力。
The full-length spike (S) gene of porcine respiratory coronavirus (PRCV) was inserted into the genome of human adenovirus type 5 downstream of the early transcription region 3 promoter. The recombinant virus replicated in cultures of the swine testicle ST cell line and directed the synthesis of S antigen with a maximum yield of approximately 26 mu g per 10(6) cells. The antigen was cell-associated except in the late phase of the infection, when a small amount(3.5 mu g per 10(6) cells) was released. The cell-associated antigen consisted of polypeptides of molecular mass 160kDa and 175kDa, comigrating with the authentic precursor S' and the mature S protein of PRCV, respectively. The extracellular recombinant antigen corresponded to the 175kDa mature protein. Some recombinant S protein was exposed on the cell surface and was recognized by neutralization-mediating anti-S monoclonal antibodies. Piglets, inoculated oronasally with the recombinant adenovirus vector developed PRCV-neutralizing serum antibodies and were partially protected against PRCV challenge, demonstrating the potential of live adenovirus as vaccine vector.