Suárez P, Díaz-Guerra M, Prieto C, Esteban M, Castro J M, Nieto A, Ortín J
Centro Nacional de Biotecnologiá, Madrid, Spain.
J Virol. 1996 May;70(5):2876-82. doi: 10.1128/JVI.70.5.2876-2882.1996.
The gene product of open reading frame 5 (p25) of porcine reproductive and respiratory syndrome (PRRS) virus has been expressed by coinfection of culture cells with vaccinia virus expressing the T7 RNA polymerase and a recombinant vaccinia virus encoding the open reading frame 5 gene under the T7 promoter and the encephalomyocarditis virus internal ribosome entry site. In spite of the reported efficiency of the expression system, very poor accumulation of p25 protein was observed and a strong cytotoxicity was produced in the doubly infected cells. This cell toxicity was shown to occur by induction of apoptosis, as indicated by nucleosome ladder formation, chromatin condensation, and rRNA degradation. Apoptosis induction was also observed after infection of cultured cells with an adapted PRRS virus strain and after infection of swine macrophage cells with a PRRS virus field strain. Contrary to the observations made for other cases of virus-induced apoptosis, we could not prevent p25 protein-induced apoptosis by using a cell line permanently expressing Bcl-2 protein.
通过将表达T7 RNA聚合酶的痘苗病毒与在T7启动子和脑心肌炎病毒内部核糖体进入位点控制下编码开放阅读框5基因的重组痘苗病毒共感染培养细胞,表达了猪繁殖与呼吸综合征(PRRS)病毒开放阅读框5的基因产物(p25)。尽管报道了该表达系统的效率,但观察到p25蛋白的积累非常少,并且在双重感染的细胞中产生了强烈的细胞毒性。这种细胞毒性表现为通过诱导凋亡发生,如核小体梯状条带形成、染色质浓缩和rRNA降解所示。在用适应性PRRS病毒株感染培养细胞后以及用PRRS病毒野毒株感染猪巨噬细胞后,也观察到了凋亡诱导。与其他病毒诱导凋亡的情况不同,我们不能通过使用永久表达Bcl-2蛋白的细胞系来预防p25蛋白诱导的凋亡。