Efficient transduction of mammalian cells by a recombinant baculovirus having the vesicular stomatitis virus G glycoprotein
Efficient transduction of mammalian cells by a recombinant baculovirus having the vesicular stomatitis virus G glycoprotein
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DOI：
10.1089/hum.1997.8.17-2011
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发表时间：
1997-11-20
影响因子：
4.2
通讯作者：
Boyce, FM
中科院分区：
文献类型：
--
作者：
Barsoum, J;Brown, R;Boyce, FM
杆状病毒载体能有效地转导培养的人肝癌细胞和原代肝细胞。本文报道了一种新的重组杆状病毒（VGZ 3）的产生，其中水泡性口炎病毒糖蛋白G（VSV G）存在于病毒包膜中，将编码VSV G的基因在多角体蛋白启动子的控制下插入杆状病毒基因组中，使得其在感染的昆虫细胞中以非常高的水平表达，但在感染的昆虫细胞中不表达。LacZ报告基因的表达由在哺乳动物细胞中有功能的启动子（Rous肉瘤病毒长末端重复序列）驱动。我们通过Western分析表明，VSV G蛋白存在于纯化的杆状病毒制剂中，VSV G单克隆抗体阻断VGZ 3对哺乳动物细胞的转导。该病毒在形态上不同于缺乏VSV G的杆状病毒，病毒粒子采用椭圆形而不是杆状形态，VGZ 3在体外转导人肝癌细胞的效率大约是缺乏VSV G的杆状病毒的10倍（病毒Z4），VGZ 3也能够转导不能被24有效转导的细胞系，我们提供的证据表明，VSV G蛋白可能通过增加杆状病毒从细胞内囊泡逃逸的效率，而不是通过增加细胞结合或病毒的摄取，从而增强转导。讨论了这种杆状病毒和相关杆状病毒在基因治疗中的可能用途。
Baculovirus vectors recently have been shown to be capable of efficient transduction of human hepatoma cells and primary hepatocytes in culture, This paper describes the generation of a novel recombinant baculovirus (VGZ3) in which the vesicular stomatitis virus glycoprotein G (VSV G) is present in the viral envelope, The gene encoding VSV G was inserted into the baculovirus genome under the control of the polyhedrin promoter such that it was expressed at very high levels in infected insect cells but not in mammalian cells, Expression of the lacZ reporter gene was driven by a promoter that is functional in mammalian cells (the Rous sarcoma virus long terminal repeat), We show by Western analysis that VSV G protein was present in purified baculovirus preparations, A VSV G monoclonal antibody blocked transduction of mammalian cells by VGZ3. This virus was morphologically distinct from baculovirus lacking VSV G, with virions adopting an oval rather than rod-shaped morphology, VGZ3 transduced human hepatoma cells in vitro at an efficiency roughly 10-fold greater than baculovirus lacking VSV G (the virus Z4), VGZ3 was also capable of transducing cell lines that could not be transduced efficiently by 24, We provide evidence that VSV G protein may enhance transduction by increasing the efficiency of escape of baculovirus from intracellular vesicles rather than by increasing cell binding or uptake of the virus, The possible use of this and related baculoviruses in gene therapy is discussed.