Yoshimatsu K, Arikawa J, Tamura M, Yoshida R, Lundkvist A, Niklasson B, Kariwa H, Azuma I
Institute for Animal Experimentation, School of Medicine, Hokkaido University, Japan.
J Gen Virol. 1996 Apr;77 ( Pt 4):695-704. doi: 10.1099/0022-1317-77-4-695.
We characterized the antigenic sites on the nucleocapsid protein (NP) of Hantaan virus (HTN) using 10 monoclonal antibodies (MAbs). At least seven antigenic sites were revealed by a competitive binding assay and divided into three partially overlapping antigenic regions (I, II and III). Regions I [amino acids (aa) 1-103], II (aa 104-204) and III (aa 205-402) were mapped on NP by examining the reactivity of truncated gene products. Those that corresponded to region I reacted with immune mouse serum, indicating that the region contained major linear epitopes as reported with Four corners virus (FCV) and Puumala virus (PUU) NP. At least one MAb to each region inhibited viral growth when they were introduced into cells by scrape-loading. In addition, they conferred protection from a lethal HTN challenge to newborn mice. A PEPSCAN assay localized the epitope of MAb E5/G6 between aa 166-175. Since E5/G6, which had the highest inhibitory effect both in cells and in mice, showed no virus neutralization activity by ordinary neutralization test, this region is suggested to be important for the virus growth after entry into the cells.
我们使用10种单克隆抗体（MAb）对汉坦病毒（HTN）核衣壳蛋白（NP）上的抗原位点进行了表征。通过竞争结合试验揭示了至少7个抗原位点，并将其分为三个部分重叠的抗原区域（I、II和III）。通过检测截短基因产物的反应性，将区域I [氨基酸（aa）1 - 103]、II（aa 104 - 204）和III（aa 205 - 402）定位在NP上。与区域I相对应的截短产物能与免疫小鼠血清反应，表明该区域含有如四角病毒（FCV）和普马拉病毒（PUU）NP所报道的主要线性表位。当通过刮涂加载将针对每个区域的至少一种MAb引入细胞时，它们会抑制病毒生长。此外，它们能保护新生小鼠免受致死性HTN攻击。PEPSCAN分析将MAb E5/G6的表位定位在aa 166 - 175之间。由于在细胞和小鼠中均具有最高抑制作用的E5/G6通过常规中和试验未显示病毒中和活性，因此该区域被认为对于病毒进入细胞后的生长很重要。
