Nagata S, Okamoto Y, Inoue T, Ueno Y, Kurata T, Chiba J
Department of Pathology, National Institute of Health, Tokyo, Japan.
Arch Virol. 1992;127(1-4):153-68. doi: 10.1007/BF01309581.
Thirteen monoclonal antibodies (MAbs) to the glycoprotein (G) of vesicular stomatitis virus (VSV) serotype Indiana were prepared and examined for their effects on various biological activities of VSV, including in vitro infection, hemagglutination, adsorption to cells, and mediation of cell fusion. Competitive binding assays with these MAbs revealed the presence of at least seven distinct antigenic determinants (epitopes) on the G protein. In some cases, overlappings among epitopes to various degrees were observed as partial inhibition or binding enhancement. The MAbs to all the epitopes but one (epitopes 1-6) reacted with the denatured G protein in a Western immunoblot analysis. Four of the epitopes (epitopes 2, 4, 5, and 7) were involved in neutralization and two (epitopes 1 and 2) in hemagglutination inhibition. None of the MAbs inhibited the adsorption of radiolabeled VSV to BHK-21 cells; the MAbs to epitope 2 slightly enhanced the virus adsorption. All neutralization epitopes except epitope 2 (epitopes 4, 5, and 7) were associated with inhibition of VSV-mediated cell fusion. These results show a direct spatial relationship between the epitopes recognized by the MAbs and functional sites on G protein and further insights into the structure and function of G protein.
制备了13种针对水疱性口炎病毒（VSV）印第安纳血清型糖蛋白（G）的单克隆抗体（MAb），并检测了它们对VSV各种生物学活性的影响，包括体外感染、血凝、细胞吸附以及细胞融合介导作用。用这些单克隆抗体进行的竞争性结合试验表明，G蛋白上至少存在7个不同的抗原决定簇（表位）。在某些情况下，观察到表位之间存在不同程度的重叠，表现为部分抑制或结合增强。除一个表位（表位1-6）外，针对所有表位的单克隆抗体在蛋白质免疫印迹分析中均与变性G蛋白发生反应。其中4个表位（表位2、4、5和7）参与中和作用，2个表位（表位1和2）参与血凝抑制作用。没有一种单克隆抗体抑制放射性标记的VSV吸附到BHK-21细胞上；针对表位2的单克隆抗体略微增强了病毒吸附。除表位2外的所有中和表位（表位4、5和7）均与抑制VSV介导的细胞融合有关。这些结果显示了单克隆抗体识别的表位与G蛋白功能位点之间的直接空间关系，并进一步深入了解了G蛋白的结构和功能。
