Replication of adeno-associated virus in synchronized cells without the addition of a helper virus
Replication of adeno-associated virus in synchronized cells without the addition of a helper virus
复制标题
腺相关病毒在同步细胞中的复制，无需添加辅助病毒
作者：
B. Yakobson;T. Koch;E. Winocour
我们研究了腺相关病毒（AAV）在细胞中通过羟基脲预处理、逆转多胺消耗或物理有丝分裂分离而同步化的辅助复制。在感染前用羟基脲处理的中国仓鼠细胞（OD4 系）中。 AAV 经历了一个完整的复制周期。用质粒克隆的 AAV DNA 转染此类细胞也产生了感染性病毒后代。通过逆转多胺耗竭或有丝分裂分离使 OD4 细胞同步化，导致独立的 AAV DNA 合成（以及前一过程中的感染性病毒后代），但这些过程不如羟基脲预处理有效。在感染前用羟基脲处理的中国仓鼠、人类和猴子来源的一些其他细胞系中也检测到独立的 AAV DNA 合成。结果表明，与之前的观点相反，AAV 感染过程并不绝对依赖于共感染辅助病毒的添加。
We investigated the helper-independent replication of adeno-associated virus (AAV) in cells synchronized by pretreatment with hydroxyurea, reversal of polyamine depletion, or physical mitotic detachment. In Chinese hamster cells (OD4 line) treated with hydroxyurea prior to infection. AAV underwent a complete cycle of replication. Transfection of such cells with plasmid-cloned AAV DNAs also gave rise to infectious viral progeny. Synchronization of OD4 cells by reversal of polyamine depletion or mitotic detachment led to independent AAV DNA synthesis (and infectious viral progeny in the case of the former procedure), but these procedures were not as effective as hydroxyurea pretreatment. Independent AAV DNA synthesis was also detected in some other cell lines of Chinese hamster, human, and monkey origin treated with hydroxyurea prior to infection. The results demonstrate that, in contrast to previous notions, the AAV infectious process is not absolutely dependent upon the addition of a coinfecting helper virus.