The Major Hurdle for Effective Baculovirus Transduction into Mammalian Cells Is Passing Early Endosomes
文献类型：期刊论文
| 作者 | Hu, Liangbo1,2; Li, Yimeng1,2; Ning, Yun-Jia1; Deng, Fei1; Vlak, Just M.3; Hu, Zhihong1; Wang, Hualin1; Wang, Manli1 |
| 刊名 | JOURNAL OF VIROLOGY
 |
| 出版日期 | 2019-08-01 |
| 卷号 | 93期号:15页码:18 |
| 关键词 | baculovirus early endosome entry fusion gene delivery mammalian cells transduction |
| ISSN号 | 0022-538X |
| DOI | 10.1128/JVI.00709-19 |
| 英文摘要 | Baculoviruses, although they infect insects in nature, can transduce a wide variety of mammalian cells and are therefore promising gene therapy vectors. However, baculovirus transduction into many mammalian cells is very inefficient, and the limiting stages and factors remain unknown. An important finding is that a short-duration trigger with low pH can significantly enhance virus transduction efficiency, but the mechanism is poorly understood. Herein, we performed a detailed comparative study on entry mechanisms of the prototypical baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) into insect and mammalian cells. The results showed that AcMNPV could be internalized into mammalian cells efficiently, but fusion in early endosomes (EEs) appeared to be the major obstacle. Measurement of endosomal pH suggested that virus fusion might be restricted under relatively high-pH conditions in mammalian cells. Interestingly, mutations of the major viral fusion protein GP64 that conferred decreased fusogenicity did not affect virus infection of insect cells, whereas virus transduction into mammalian cells was severely impaired, suggesting a more stringent dependence on GP64 fusogenicity for AcMNPV entry into mammalian cells than into insect cells. An increase in the fusogenicity of GP64 mutants resulting from low pH triggered the rescue of fusion-deficient recombinant virus transduction efficiency. Based on the above-described findings, the pH of EEs was specifically reduced with a Na+/K+-ATPase inhibitor, and the AcMNPV transduction of many mammalian cells indeed became highly efficient. This study not only revealed the roadblocks to mammalian cell entry of baculovirus but also provides a new strategy for improving baculovirus-based gene delivery and therapy. IMPORTANCE Baculoviruses can transduce a wide variety of mammalian cells but do so with low efficiency, which greatly limits their practical application as potential gene delivery vectors. So far, the understanding of baculovirus entry into mammalian cells is obscure, and the limiting stages and factors are unclear. In this study, by comparatively analyzing the mechanisms of baculovirus entry into mammalian and insect cells, virus fusion during the early stage of endocytosis was revealed as the major obstacle for efficient baculovirus transduction into mammalian cells. A higher fusogenicity of the major viral fusion protein GP64 was found to be required for virus entry into mammalian cells than for entry into insect cells. Interestingly, by decreasing the pH of early endosomes with a specific agent, virus transduction of a wide range of mammalian cells was greatly enhanced. This study uncovers the roadblocks to mammalian cell entry of baculoviruses and presents mechanisms to overcome the roadblocks. |
| 资助项目 | National Natural Science Foundation of China[31621061] ; National Natural Science Foundation of China[31370191] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDB11030400] ; Wuhan Institute of Virology ; State Key Laboratory of Virology |
| WOS研究方向 | Virology |
| 语种 | 英语 |
| WOS记录号 | WOS:000475793200029 |
| 出版者 | AMER SOC MICROBIOLOGY |
| 源URL | [http://202.127.146.157/handle/2RYDP1HH/8363]  |
| 专题 | 中国科学院武汉植物园 |
| 通讯作者 | Wang, Manli |
| 作者单位 | 1.Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan, Hubei, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.Wageningen Univ & Res, Lab Virol, Wageningen, Netherlands |
| 推荐引用方式 GB/T 7714 | Hu, Liangbo,Li, Yimeng,Ning, Yun-Jia,et al. The Major Hurdle for Effective Baculovirus Transduction into Mammalian Cells Is Passing Early Endosomes[J]. JOURNAL OF VIROLOGY,2019,93(15):18. |
| APA | Hu, Liangbo.,Li, Yimeng.,Ning, Yun-Jia.,Deng, Fei.,Vlak, Just M..,...&Wang, Manli.(2019).The Major Hurdle for Effective Baculovirus Transduction into Mammalian Cells Is Passing Early Endosomes.JOURNAL OF VIROLOGY,93(15),18. |
| MLA | Hu, Liangbo,et al."The Major Hurdle for Effective Baculovirus Transduction into Mammalian Cells Is Passing Early Endosomes".JOURNAL OF VIROLOGY 93.15(2019):18. |
入库方式： OAI收割
来源：武汉植物园
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。