Steimer K S, Boettiger D
J Virol. 1979 Oct;32(1):175-86. doi: 10.1128/JVI.32.1.175-186.1979.
Procedures for characterizing replication-defective viruses in nonpermissive mammalian cells were developed and applied to three nonvirogenic Rous sarcoma virus (RSV)-transformed mammalian cell lines--B4, a line of Bryan virus-transformed hamster cells, and two SRD-RSV transformed rat cell lines, LR3/1 and LR3/2. Cell fusion was used to study virus complementation. The three cell lines (i) fused with helper virus-infected chicken cells and the host range of the rescued virus examined, (ii) tested for complementation by fusion with chicken cells exhibiting various patterns of endogenous virus expression, (iii) fused with chicken cells infected with the temperature-sensitive replication mutant LA334 and assayed for complementation at permissive and nonpermissive temperatures, and (iv) tested for complementation of defective viruses in other RSV-transformed mammalian cell lines by fusing pairs of nonvirogenic cell lines and permissive chicken cells. Based upon these complementation studies, we concluded that B4 virus is defective only in the env gene, LR3/) virus is an absolute mutant in the gag and/or pol genes, and LR3/2 virus is a leaky env mutant. Clones of LR3/1 and LR3/2 virus-infected chicken cells were established, and the results obtained from the characterization of these viruses in permissive avian cells substantiates the conclusions reached in the fusion-rescue studies.
已开发出在非允许性哺乳动物细胞中鉴定复制缺陷型病毒的程序,并将其应用于三种非致瘤性劳氏肉瘤病毒(RSV)转化的哺乳动物细胞系——B4,一种布莱恩病毒转化的仓鼠细胞系,以及两种SRD-RSV转化的大鼠细胞系LR3/1和LR3/2。细胞融合被用于研究病毒互补作用。这三种细胞系:(i)与辅助病毒感染的鸡细胞融合,并检测拯救病毒的宿主范围;(ii)通过与呈现各种内源性病毒表达模式的鸡细胞融合来检测互补作用;(iii)与感染温度敏感型复制突变体LA334的鸡细胞融合,并在允许温度和非允许温度下检测互补作用;(iv)通过将非致瘤性细胞系对与允许性鸡细胞融合,检测其他RSV转化的哺乳动物细胞系中缺陷病毒的互补作用。基于这些互补作用研究,我们得出结论:B4病毒仅在env基因中有缺陷,LR3/1病毒在gag和/或pol基因中是绝对突变体,而LR3/2病毒是一种渗漏型env突变体。建立了感染LR3/1和LR3/2病毒的鸡细胞克隆,并且在允许性禽类细胞中对这些病毒进行鉴定所获得的结果证实了融合拯救研究中得出的结论。