Efficient delivery and stable gene expression in a hematopoietic cell line using a chimeric serotype 35 fiber pseudotyped helper-dependent adenoviral vector
Efficient delivery and stable gene expression in a hematopoietic cell line using a chimeric serotype 35 fiber pseudotyped helper-dependent adenoviral vector
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DOI：
10.1016/j.virol.2004.03.030
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发表时间：
2004-06-20
期刊：
影响因子：
3.7
通讯作者：
Mitani, K
中科院分区：
文献类型：
--
作者：
Balamotis, MA;Huang, K;Mitani, K
由于缺乏柯萨奇病毒 B 腺病毒受体 (CAR)，某些人类细胞群体仍然难以被人类腺病毒 (Ad) 血清型 5 感染。天然腺病毒纤维成分虽然多种多样，但不能感染所有组织类型。最近，创建了一种嵌合 Ad5/35 纤维，它显示出与 Ad5 不同的向性。我们将这种嵌合纤维整合到辅助依赖性（HD）腺病毒载体系统中，并通过转基因表达、细胞转导效率和细胞毒性比较HD与E1缺失（E1Delta）载体。与 Ad5 纤维相比，嵌合 Ad5/35 纤维感染 K562 细胞的效率高出 50 倍。在体外感染活跃分裂的 K562 细胞后，HD Ad5/35 的短期转基因表达比 E1Delta Ad5/35 载体的持续时间更长。通过定量实时 PCR (QRT-PCR) 或细胞毒性确定，E1Delta Ad5/35 感染引起的转基因表达快速丧失并不是载体基因组丧失的线索，而是通过假定的沉默机制确定的。 (C) 2004 Elsevier Inc. 保留所有权利。
Certain human cell Populations have remained difficult to infect with human adenovirus (Ad) serotype 5 because of their lack of coxsackievirus B-adenovirus receptor (CAR). Native adenovirus fiber compositions, although diverse, cannot infect all tissue types. Recently, a chimeric Ad5/35 fiber was created, which displays an altered tropism from Ad5. We incorporated this chimeric fiber into a helper-dependent (HD) adenovirus vector system and compared HD to E1-deleted (E1Delta) vectors by transgene expression, cell transduction efficiency, and cytotoxicity. K562 cells were infected similar to50 times more efficiently with the chimeric Ad5/35 fiber compared with the Ad5 fiber. Short-term transgene expression was sustained longer from HD Ad5/35 than E1Delta Ad5/35 vector after in vitro infection of actively dividing K562 cells. Rapid loss of transgene expression from E1Delta Ad5/35 infection was not clue to the loss of vector genomes, as determined by quantitative real-time PCR (QRT-PCR), or cytotoxicity, but rather through a putative silencing mechanism. (C) 2004 Elsevier Inc. All rights reserved.