Lundstrom K
F. Hoffman-La Roche, Research Laboratories, Basel, Switzerland.
J Recept Signal Transduct Res. 1999 Jan-Jul;19(1-4):673-86. doi: 10.3109/10799899909036679.
The broad host range and superior infectivity of alphaviruses have encouraged the development of efficient expression vectors for Semliki Forest virus (SFV) and Sindbis virus (SIN). The generation of high-titer recombinant alphavirus stocks has allowed high-level expression of a multitude of nuclear, cytoplasmic, membrane-associated and secreted proteins in a variety of different cell lines and primary cell cultures. Despite the viral cytopathogenic effects, functional assays on recombinant proteins are possible for a time-period of at least 24 hours post-infection. The high percentage (80-95%) of primary neurons infected with SFV has allowed localization and functional studies of recombinant proteins in these primary cell cultures. Through multiple infection studies the interaction of receptor and G protein subunits has become feasible. Establishment of efficient scale-up procedures has allowed production of large quantities of recombinant protein. Potential gene therapy applications of alphaviruses could be demonstrated by injection of recombinant SIN particles expressing beta-galactosidase into mouse brain. Tissue/cell specific infection has been achieved by introduction of an IgG-binding domain of protein A domain into one of the spike proteins of SIN. This enabled efficient targeting of infection to human lymphoblastoid cells.
甲病毒广泛的宿主范围和卓越的感染性推动了针对辛德毕斯病毒（SIN）和Semliki森林病毒（SFV）的高效表达载体的开发。高滴度重组甲病毒毒株的产生使得众多核蛋白、胞质蛋白、膜相关蛋白和分泌蛋白能够在多种不同的细胞系和原代细胞培养物中实现高水平表达。尽管存在病毒致细胞病变效应，但在感染后至少24小时的时间段内仍可对重组蛋白进行功能测定。感染SFV的原代神经元比例很高（80 - 95%），这使得在这些原代细胞培养物中对重组蛋白进行定位和功能研究成为可能。通过多次感染研究，受体与G蛋白亚基的相互作用变得可行。高效放大生产程序的建立使得能够大量生产重组蛋白。将表达β - 半乳糖苷酶的重组SIN颗粒注射到小鼠脑内，可证明甲病毒在基因治疗方面的潜在应用。通过将蛋白A结构域的IgG结合结构域引入SIN的一种刺突蛋白中，实现了组织/细胞特异性感染。这使得能够有效地将感染靶向人淋巴母细胞。
