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
作者：邱薇(成都军区疾病预防控制中心)；王文博(成都军区疾病预防控制中心)；胡挺松(成都军区疾病预防控制中心)；郭平(成都军区疾病预防控制中心)；周卫国(成都军区疾病预防控制中心)；张国富(成都军区疾病预防控制中心)；郑颖(成都军区疾病预防控制中心)；范泉水(成都军区疾病预防控制中心)
Author：QIU Wei(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)；WANG Wen-Bo(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)；HU Ting-Song(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)；GUO Ping(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)；ZHOU Wei-Guo(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)；ZHANG Guo-Fu(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)；ZHENG Ying(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)；FAN Quan-Shui(Center for Disease Control and Prevention, Chinese PLA Cheng Du Military Command)
收稿日期：2016-11-15 年卷（期）页码：2017,54(5):1089-1093
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：恒河猴；犬瘟热病毒；基因疫苗；单克隆抗体
Key words：rhesus monkey ; Canine Distemper virus; Gene vaccine; Monoclonal antibody
基金项目：国家公益性行业（农业）科研重大专项（201303042）
中文摘要
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。
英文摘要
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.
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