Miller D G, Adam M A, Miller A D
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Mol Cell Biol. 1990 Aug;10(8):4239-42. doi: 10.1128/mcb.10.8.4239-4242.1990.
Previous reports have shown that retrovirus infection is inhibited in nonreplicating (stationary-phase [hereafter called stationary]) cells. Infection of stationary cells was shown to occur when the cells were allowed to replicate at times up to a week after infection, suggesting that an unintegrated retrovirus could persist in a form that was competent to integrate after release of the block to replication. However, those studies were complicated by the use of replication-competent virus, which can spread in the infected cells. We have used a replication-defective retrovirus vector to compare the efficiency of gene transfer in stationary and replicating rat embryo fibroblasts. In agreement with previous results, gene transfer was inhibited 100-fold in stationary versus replicating cells. In contrast to previously reported results, the block to infection could not be relieved by stimulating stationary cells to divide at times from 6 h to 10 days after infection. Thus, for successful retroviral infection, the infected cells must be replicating at the time of infection. These results have important implications for the use of retroviral vectors for gene transfer.
先前的报道显示，逆转录病毒感染在非复制型（静止期[以下简称静止期]）细胞中受到抑制。当静止期细胞在感染后长达一周的时间内被允许复制时，显示会发生感染，这表明未整合的逆转录病毒能够以一种在解除复制阻滞后有能力整合的形式持续存在。然而，那些研究因使用了具有复制能力的病毒而变得复杂，这种病毒能够在被感染的细胞中传播。我们使用了一种复制缺陷型逆转录病毒载体来比较静止期和复制期大鼠胚胎成纤维细胞中的基因转移效率。与先前的结果一致，静止期细胞与复制期细胞相比，基因转移受到100倍的抑制。与先前报道的结果相反，在感染后6小时至10天的任何时间刺激静止期细胞分裂，都无法解除感染阻滞。因此，为了实现成功的逆转录病毒感染，被感染的细胞在感染时必须处于复制状态。这些结果对于使用逆转录病毒载体进行基因转移具有重要意义。
