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Studies on Epstein-Barr virus vector and human gene therapy

EB病毒载体及人类基因治疗的研究

基本信息

批准号：
07044272
负责人：
TAKADA Kenzo
金额：
$ 3.78万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07044272/
关键词：
Epstein-Barr virus Gene therapy Viral vector

项目摘要

There has been no infection system such as that which produces viruses following Epstein-Barr virus (EBV) infection. Therefore, it has not been possible to produce a large amount of recombinant EBV for human gene therapy. We have recently established a system for producing recombinant EBV.When considering to use the EBV vector for application to human, we need to develop the EBV vector with deletions of transforming genes. In the present study, following experiments were carried out.1. Generation of EBV recombinants deleted of the latent membrane protein, which play crucial roles in lymphocyte transformation.2. Generation of EBV recombinants incompetent for virus replication and packaging cells for their propagation :3. Generation of the amplicon system : DNA sequences containing the replication origin in a virus replicative phase and packaging signals are efficiently amplified and packaged into virus particles during virus replication. We are now trying to create cells infected with recombinant EBV with more than 200 kbp genome size. Such large EBV genomes exceed the size being packaged into virus particles. Therefore, such cells should become an ideal packaging cells for propagating amplicon DNA.Dr.Bill Sugden visited our laboratory in February, 1997. Dr.Sugiura visited Dr.Sugden's laboratory in January, 1997. Drs.Takada and Imai visited Dr.Sugden's laboratory in March, 1997. Through these communication, we discussed about mutual experimental results, and obtained many important suggestions each other.

没有感染系统，如在EB病毒（EBV）感染后产生病毒的系统。因此，不可能生产大量用于人基因治疗的重组EBV。我们最近建立了一个生产重组EBV的系统。当考虑将EBV载体应用于人类时，我们需要开发具有转化基因缺失的EBV载体。在本研究中，进行了以下实验.目的：1.构建在淋巴细胞转化中起关键作用的潜伏膜蛋白缺失的EBV重组体.产生不能进行病毒复制的EBV重组体和用于其增殖的包装细胞：3.扩增子系统的产生：在病毒复制过程中，含有病毒复制期复制起点和包装信号的DNA序列被有效扩增并包装到病毒颗粒中。我们现在正试图创建用基因组大小超过200 kbp的重组EBV感染的细胞。如此大的EBV基因组超过了包装成病毒颗粒的大小。因此，这种细胞应该成为繁殖扩增子DNA的理想包装细胞。Bill Sugden博士于1997年2月访问了我们的实验室。杉浦博士于1997年1月访问了萨格登博士的实验室。高田博士和今井博士于1997年3月访问了萨格登博士的实验室。通过这些交流，我们对相互的实验结果进行了讨论，并得到了许多重要的建议。