Jiang W, Venugopal K, Gould E A
Natural Environment Research Council Institute of Virology and Environmental Microbiology, Oxford, United Kingdom.
J Virol. 1995 Feb;69(2):1044-9. doi: 10.1128/JVI.69.2.1044-1049.1995.
A single-chain antibody fragment that identifies a neutralizing epitope on the envelope protein of louping ill and some other tick-borne flaviviruses was previously expressed in soluble form from bacteria and shown to be functionally active in vitro. To see whether or not the single-chain antibody could bind and inactivate infectious virus in vivo, we have used recombinant Sindbis virus as a delivery vehicle for intracellular expression of the antibody fragment. The variable genes and interchain linker encoding the single-chain antibody were cloned into a double subgenomic Sindbis virus expression vector to generate recombinant Sindbis virus. Infection with this recombinant Sindbis virus provided high-level cytoplasmic expression of the antibody fragment in mammalian cells. We demonstrate (i) that the antibody fragment was antigen binding and (ii) that louping ill virus infectivity was significantly reduced in the presence of intracellular antibody expressed by the superinfecting recombinant Sindbis virus.
一种能识别跳跃病及其他一些蜱传黄病毒包膜蛋白上中和表位的单链抗体片段，此前已在细菌中以可溶形式表达，并在体外显示具有功能活性。为了观察该单链抗体在体内是否能够结合并灭活感染性病毒，我们使用重组辛德毕斯病毒作为载体在细胞内表达抗体片段。将编码单链抗体的可变基因和链间连接子克隆到双亚基因组辛德毕斯病毒表达载体中，以产生重组辛德毕斯病毒。用这种重组辛德毕斯病毒感染可使抗体片段在哺乳动物细胞中高水平地在细胞质中表达。我们证明：（i）抗体片段具有抗原结合能力；（ii）在超感染的重组辛德毕斯病毒表达的细胞内抗体存在的情况下，跳跃病病毒的感染性显著降低。
