Deng Yong-Ning, Zeng Jun-Yan, Su Hua, Qu Qiu-Min
Department of Neurology, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Intervirology. 2015;58(6):363-8. doi: 10.1159/000443832. Epub 2016 Feb 25.
Virus-like particle (VLP) of JC polyomavirus (JCPyV) is capable of packaging and delivering exogenous DNA into human cells and can be used for mediating therapeutic gene expression. However, many human cells express the JCPyV receptor. Thus, wild-type VLP can transduce a wide range of human cells nonspecifically. This study tested the feasibility of using a modified VLP with a IgG binding domain (Z domain) of protein A in its capsid for targeted gene delivery. The Z domain of protein A isolated from the membrane of Staphylococcus aureus was inserted into the NH3-terminus of VP1, the major JCPyV capsular protein. The recombinant VLP-Z was produced using Escherichia coli. Electron-microscopic analysis showed that VLP-Z has a viral-like structure. A hemagglutination test showed that VLP-Z has hemagglutination activity. VP(1) was detected in the nuclei of HeLa cells by immunostaining after VLP-Z inoculation, suggesting that VLP-Z is viable and can enter the cell nucleus. Inoculating HeLa cells with pEGFP-N(1) plasmid packaged in VLP-Z has resulted in enhanced green fluorescent protein expression in the cells. In addition, a binding assay showed that VLP-Z was able to bind IgG through the interaction of the Z domain in VLP-Z and IgG. These data suggest that VLP-Z could be armed with cell-specific antibody and be used to deliver therapeutic genes to target cells.
JC多瘤病毒（JCPyV）的病毒样颗粒（VLP）能够将外源性DNA包装并递送至人类细胞，可用于介导治疗性基因表达。然而，许多人类细胞表达JCPyV受体。因此，野生型VLP可非特异性转导多种人类细胞。本研究测试了在衣壳中使用带有蛋白A的IgG结合结构域（Z结构域）的修饰VLP进行靶向基因递送的可行性。从金黄色葡萄球菌膜中分离的蛋白A的Z结构域被插入到主要的JCPyV衣壳蛋白VP1的NH3末端。使用大肠杆菌生产重组VLP-Z。电子显微镜分析表明VLP-Z具有病毒样结构。血凝试验表明VLP-Z具有血凝活性。VLP-Z接种后通过免疫染色在HeLa细胞核中检测到VP(1)，表明VLP-Z是有活力的并且能够进入细胞核。用包装在VLP-Z中的pEGFP-N(1)质粒接种HeLa细胞导致细胞中绿色荧光蛋白表达增强。此外，结合试验表明VLP-Z能够通过VLP-Z中的Z结构域与IgG的相互作用结合IgG。这些数据表明VLP-Z可以携带细胞特异性抗体并用于将治疗性基因递送至靶细胞。
