PASSIVE IMMUNIZATION--TREATING RABIES WITH HUMAN MAB

该项目的目标是产生狂犬病病毒中和 人源单抗(MAbs)，优先选用高亲和力的抗体 同种类型，因此几种中和人源单抗的“鸡尾酒” 针对病毒糖蛋白(GP)上多个抗原位点的制备 和核糖核蛋白(RNP)复合体。具体地说，该项目旨在： (L)产生至少15株高亲和力、中和病毒的人源单抗 用人抗狂犬病病毒不同抗原决定簇 杂交瘤技术和(2)表明这些单抗可以保护动物免受 致命的狂犬病挑战。从外周血中分离出B细胞(10-14 在加强疫苗接种后)将因感染EBV而永垂不朽， 以及在培养中检测到的产生狂犬病病毒特异性抗体的细胞 通过酶联免疫吸附试验，将上清液融合成人鼠杂交种。 这些杂交细胞将被克隆，并产生狂犬病病毒特异性的细胞 抗体扩增。以这种方式产生的狂犬病病毒特异性单抗 将在体外充分鉴定同种异型，特异性，亲和力， 和VNA滴度和体内暴露后效应的小鼠和 叙利亚仓鼠模型。

The objective of this project is to generate rabies virus neutralizing human monoclonal antibody (mAbs), preferentially of the high affinity IgG isotype, so that a "cocktail" of several neutralizing human mAbs can be prepared against multiple antigenic sites on the viral glycoprotein (GP) and ribonucleoprotein (RNP) complex. Specifically, this project aims to: (l) generate at least 15 high-affinity, virus-neutralizing human mAbs against various antigenic determinants on rabies virus using human hybridoma technology and (2) show that these mAbs protect animals against lethal rabies challenge. B cells isolated from peripheral blood (10-14 days) after booster vaccine) will be immortalized by infection with EBV, and cells producing rabies virus-specific antibody, detected in culture supernatants by ELISA, will be fused to produce mouse-human hybrids. These hybrids will be cloned, and cells producing rabies virus-specific antibody expanded. Rabies virus-specific mAbs generated in this fashion will be fully characterized in vitro for isotype, specificity, affinity, and VNA titre and in vivo for post-exposure efficacy in mice and the Syrian hamster model.