Baculovirus GP64-pseudotyped HIV-based lentivirus vectors are stabilized against complement inactivation by codisplay of decay accelerating factor (DAF) or of a GP64-DAF fusion protein
Baculovirus GP64-pseudotyped HIV-based lentivirus vectors are stabilized against complement inactivation by codisplay of decay accelerating factor (DAF) or of a GP64-DAF fusion protein
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DOI：
10.1016/j.ymthe.2004.12.002
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发表时间：
2005-04-01
影响因子：
12.4
通讯作者：
Friedmann, T
中科院分区：
文献类型：
--
作者：
Guibinga, GH;Friedmann, T
用于体内基因递送的有效基因转移载体的最终开发将需要开发许多重要的新技术，例如针对破坏或以其他方式抑制外源感染因子（如基因转移载体）的保护机制的载体稳定化。已知苜蓿银纹夜蛾核型多角体病毒的杆状病毒包膜蛋白GP 64可以有效地假型化慢病毒载体，并且杆状病毒包膜蛋白GP 64的修饰形式也可以有效地组装成杆状病毒颗粒以在杆状病毒表面上展示功能性外源蛋白。在本研究中，我们结合这些技术，制备基于HIV的慢病毒载体假型与GP 64包膜蛋白和共表达的融合蛋白的GP 64与补体调节，衰变加速因子（CD 55）。此外，我们还制备了GP 64-假型载体，其中存在一个GP 64表达质粒，以允许将GP 64蛋白掺入病毒颗粒。我们的研究结果证明了GP 64/GP 64-DAF和GP 64/DAF假型颗粒的有效表达和高滴度生产以及它们对人和非人灵长类动物血清灭活的稳定性。
The eventual development of efficient gene transfer vectors for in vivo gene delivery will require the development of a number of important new technologies such as stabilization of vectors against protective mechanisms that destroy or otherwise inactivate foreign infectious agents like gene transfer vectors. It is known that the baculovirus envelope protein GP64 of Autographa califomica nucleopolyhedrovirus can efficiently pseudotype lentivirus vectors and that modified forms of the baculovirus envelope protein GP64 can also assemble efficiently into baculovirus particles to display functional foreign proteins on the baculovirus surface. In the present study we have combined these techniques to prepare HIV-based lentivirus vectors pseudotyped with GP64 envelope protein and coexpressing a fusion protein of GP64 with the complement-regulatory, decay accelerating factor (DAF, CD55). In addition, we have also prepared GP64-pseudotyped vectors in the presence of a DAF expression plasmid to allow the incorporation of DAF protein into viral particles. Our results demonstrate both the efficient expression and the high-titer production of GP64/GP64-DAF and GP64/DAF-pseudotyped particles and their stability against inactivation by human and nonhuman primate serum.