EVIDENCE FROM THE ANTIIDIOTYPIC NETWORK THAT THE ACETYLCHOLINE-RECEPTOR IS A RABIES VIRUS RECEPTOR
EVIDENCE FROM THE ANTIIDIOTYPIC NETWORK THAT THE ACETYLCHOLINE-RECEPTOR IS A RABIES VIRUS RECEPTOR
复制标题
DOI：
10.1128/jvi.67.1.530-542.1993
复制
发表时间：
1993-01-01
影响因子：
5.4
通讯作者：
TIGNOR, GH
中科院分区：
文献类型：
--
作者：
HANHAM, CA;ZHAO, F;TIGNOR, GH
我们已经开发了独特型抗独特型单克隆抗体，为狂犬病病毒结合乙酰胆碱受体（AChR）提供了证据。用狂犬病病毒CVS株免疫BALB/c小鼠，取脾细胞与NS-1骨髓瘤细胞融合，获得杂交瘤细胞株7.12和7.25。抗体7.12与病毒糖蛋白反应，中和病毒在体内的感染性。当使用表达神经元AChR的PC 12细胞，而不使用不具有AChR的CER细胞或神经母细胞瘤细胞（克隆N18）时，它还在体外中和感染性。抗体7.25与核衣壳蛋白反应。用7.12 Fab免疫小鼠，取脾细胞与NS-1细胞融合，制备抗独特型单克隆抗体B 9。在酶联免疫吸附试验和免疫沉淀中，B 9与7.12、多克隆狂犬病毒免疫犬血清和纯化的AChR反应。AChR可抑制B_9与7.12及免疫犬血清的结合。B 9在体内外均能抑制7.12与狂犬病毒的结合。间接免疫荧光显示，B 9反应在小鼠组织的神经肌肉接头。B 9还与小鼠和猴脑组织中的不同神经元以及PC 12细胞进行间接免疫荧光反应。狂犬病毒感染小鼠脑组织中神经元成分的B 9染色大大减少。狂犬病病毒抑制B 9与PC 12细胞的结合。用B 9免疫小鼠产生低滴度的狂犬病毒中和抗体。保护这些小鼠免受致死性肌内狂犬病病毒攻击。相反，抗核衣壳抗体7.25的抗独特型抗体不与AChR反应。
We have developed idiotype-anti-idiotype monoclonal antibodies that provide evidence for rabies virus binding to the acetylcholine receptor (AChR). Hybridoma cell lines 7.12 and 7.25 resulted after fusion of NS-1 myeloma cells with spleen cells from a BALB/c mouse immunized with rabies virus strain CVS. Antibody 7.12 reacted with viral glycoprotein and neutralized virus infectivity in vivo. It also neutralized infectivity in vitro when PC12 cells, which express neuronal AChR, but not CER cells or neuroblastoma cells (clone N18), which have no AChR, were used. Antibody 7.25 reacted with nucleocapsid protein. Anti-idiotypic monoclonal antibody B9 was produced from fusion of NS-1 cells with spleen cells from a mouse immunized with 7.12 Fab. In an enzyme-linked immunosorbent assay and immunoprecipitation, B9 reacted with 7.12, polyclonal rabies virus immune dog serum, and purified AChR. The binding of B9 to 7.12 and immune dog serum was inhibited by AChR. B9 also inhibited the binding of 7.12 to rabies virus both in vitro and in vivo. Indirect immunofluorescence revealed that B9 reacted at neuromuscular junctions of mouse tissue. B9 also reacted in indirect immunofluorescence with distinct neurons in mouse and monkey brain tissue as well as with PC12 cells. B9 staining of neuronal elements in brain tissue of rabies virus-infected mice was greatly reduced. Rabies virus inhibited the binding of B9 to PC12 cells. Mice immunized with B9 developed low-titer rabies virus-neutralizing antibody. These mice were protected from lethal intramuscular rabies virus challenge. In contrast, anti-idiotypic antibody raised against nucleocapsid antibody 7.25 did not react with AChR.