恒河猴犬瘟热病毒基因疫苗制备单克隆抗体的研究

Preparation of monoclonal antibody against canine distemper virus of rhesus monkey using gene vaccine

摘要:

从感染犬瘟热病毒死亡的恒河猴肝脏组织中提取总RNA，经RT-PCR扩增H全基因并克隆到真核表达质粒pVAX中，构建了基因疫苗pVAX-H。用制备的基因疫苗免疫BaLb/c小鼠，小鼠可以产生免疫应答，免疫鼠脾细胞与SP2/0骨髓瘤细胞融合后，经间接ELISA法筛选，获得5株稳定分泌抗CDV H蛋白的单克隆抗体的杂交瘤细胞株。该5株单抗对应不同的病毒抗原表位，且均不与CPV和MV发生反应， McAb诱生的腹水可体外中和犬瘟热病毒，其中2株中和效价大于1:256。结果表明，CDV H基因疫苗可制备具有一定中和效价的抗CDV的单克隆抗体。

Abstract:

To prepare the monoclonal antibody of Canine distemper virus which infect rhrsus monkey, the H gene of the virus was amplified by RT-PCR and cloned into pVAX. The DNA vaccine pVAX-H was injected into Balb/c mice and induced a strong immunity. The spleen cells were fused with SP2/0 cells and 5 hybridoma strains secreting monoclonal antibodies against CDV H gene were achieved. The five different McAbs didn’t react with CPV and MV but could neutralize CDV in vitro. Two of them had high neutralization antibody above 1:256. The result import that the CDV H gene vaccine can be used to prepare monoclonal antibody.