Replication-deficient human adenovirus type 35 vectors for gene transfer and vaccination:: Efficient human cell infection and bypass of preexisting adenovirus immunity
Replication-deficient human adenovirus type 35 vectors for gene transfer and vaccination:: Efficient human cell infection and bypass of preexisting adenovirus immunity
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DOI：
10.1128/jvi.77.15.8263-8271.2003
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发表时间：
2003-08-01
影响因子：
5.4
通讯作者：
Havenga, M
中科院分区：
文献类型：
--
作者：
Vogels, R;Zuijdgeest, D;Havenga, M
复制缺陷型人 5 型腺病毒 (Ad5) 可以在补充细胞系（例如 PER.C6）中产生高滴度，并广泛用作疫苗和基因治疗载体。然而，预先存在的针对 Ad5 的免疫力阻碍了基因转移、免疫反应和载体介导的毒性的一致性。我们报告了人类 Ad35 被鉴定为一种全球流行率较低的病毒，并生成了易于插入异源基因的 Ad35 载体质粒系统。此外，我们还鉴定了 Ad35-E1B 区域的最小序列（分子量，55,000 [55K]），对于 PER.C6 细胞上完全缺乏 E1 的 Ad35 载体的互补至关重要。将 55K 序列稳定插入 PER.C6 细胞后，获得了有效转补 Ad5 和 Ad35 载体的细胞系 (PER.C6/55K)。我们进一步证明，Ad35 的转导不会受到预先存在的 Ad5 免疫的阻碍，并且与 Ad5 相比，Ad35 能有效感染树突状细胞、平滑肌细胞和滑膜细胞。
Replication-deficient human adenovirus type 5 (Ad5) can be produced to high titers in complementing cell lines, such as PER.C6, and is widely used as a vaccine and gene therapy vector. However, preexisting immunity against Ad5 hampers consistency of gene transfer, immunological responses, and vector-mediated toxicities. We report the identification of human Ad35 as a virus with low global prevalence and the generation of an Ad35 vector plasmid system for easy insertion of heterologous genes. In addition, we have identified the minimal sequence of the Ad35-E1B region (molecular weight, 55,000 [55K]), pivotal for complementation of fully E1-lacking Ad35 vector on PER.C6 cells. After stable insertion of the 55K sequence into PER.C6 cells a cell line was obtained (PER.C6/55K) that efficiently transcomplements both Ad5 and Ad35 vectors. We further demonstrate that transduction with Ad35 is not hampered by preexisting Ad5 immunity and that Ad35 efficiently infects dendritic cells, smooth muscle cells, and synoviocytes, in contrast to Ad5.