Ghiasi H, Nesburn A B, Kaiwar R, Wechsler S L
Ophthalmology Research, Cedars-Sinai Medical Center, Los Angeles, California.
Arch Virol. 1991;121(1-4):163-78. doi: 10.1007/BF01316752.
The DNA sequence encoding the complete herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) was inserted into a baculovirus transfer vector under control of the polyhedrin gene promoter of the baculovirus Autographa california nuclear polyhedrosis virus (AcNPV). After co-transfection of Spodoptera frugiperda (Sf9) insect cells with wild-type AcNPV DNA and the recombinant transfer vector DNA, polyhedrin-negative recombinants that expressed high levels of HSV-1 gD were isolated using immunoaffinity selection with antibody coated magnetic particles followed by plaque purification. These recombinant baculoviruses expressed a protein that was slightly smaller than virion HSV-1 gD made in Vero cells. This recombinant protein was expressed at high levels. The expressed protein was glycosylated, was found on the membrane of Sf9 cells, and reacted with gD specific antibodies. Antibodies raised in mice to the recombinant gD neutralized HSV-1 as measured by plaque reduction assays. Mice inoculated with the recombinant baculovirus were completely protected from lethal challenge with HSV-1.
将编码完整单纯疱疹病毒1型（HSV-1）糖蛋白D（gD）的DNA序列插入杆状病毒转移载体中，该载体受杆状病毒苜蓿银纹夜蛾核型多角体病毒（AcNPV）多角体蛋白基因启动子的控制。将野生型AcNPV DNA与重组转移载体DNA共转染草地贪夜蛾（Sf9）昆虫细胞后，使用包被抗体的磁性颗粒进行免疫亲和选择，随后进行噬斑纯化，分离出表达高水平HSV-1 gD的多角体蛋白阴性重组体。这些重组杆状病毒表达的一种蛋白质比在Vero细胞中产生的病毒粒子HSV-1 gD略小。这种重组蛋白高水平表达。表达的蛋白进行了糖基化修饰，存在于Sf9细胞的膜上，并与gD特异性抗体发生反应。通过噬斑减少试验测定，用重组gD免疫小鼠产生的抗体可中和HSV-1。接种重组杆状病毒的小鼠完全受到保护，免受HSV-1的致死性攻击。
