抗人类疱疹病毒6型南京分离株E5单克隆抗体的制备与鉴定  

Preparation and characterization of monoclonal antibodies against human herpesvirus 6

作　　者：姜兰兰[1] 姚堃[1] 尹全章[1] 茌静[1] 周锋[1] 卢士强[1] 刘英霞[1] 

机构地区：[1]南京医科大学微生物与免疫学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2008年第4期416-419,428,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助项目(3077196)

摘　　要：目的:制备并鉴定人疱疹病毒6型(HHV-6)单克隆抗体。方法:用纯化的HHV-6南京分离株E5免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0融合,经间接ELISA法筛选,间接免疫荧光法、免疫印迹实验等方法鉴定单抗。并采用套式PCR法和ELISA法,在口腔癌中进行了初步的应用。结果:获得了一株分泌抗HHV-6单克隆抗体的杂交瘤细胞系,命名为JA9。单克隆抗体亚类鉴定表明:其为IgG1亚类,轻链为κ。用间接ELISA法检测细胞上清单抗效价为1∶500;单抗腹水滴度为1∶8.0×104。免疫印迹实验结果表明:JA9单抗腹水与约75kuHHV-6E5病毒蛋白结合。口腔癌患者唾液的HHV-6阳性检测率结果分别是:套式PCR法60%,ELISA法40%。结论:成功制备并鉴定一株抗HHV-6南京分离株E5单克隆抗体,为HHV-6进一步的基础研究及临床快速诊断提供可能。Objective：Preparation and characterization of monoclonal antibodies against human herpesvirus 6（HHV-6）. Methods： Purified HHV-6 was immunized to BALB/c mouse,and the spleen cells of the immunized mouse were fused with SP2/0 myeloma cells. The positive clones which produced McAbs against HHV-6 were screened by ELISA,indirect immunofluorescence assay. And samples of saliva from 25 oral squamous cell carcinoma patients were tested for human herpesvirus 6 by nested PCR assay and ELISA. Results：Clones named as JA9 could secrete specific McAbs against HHV-6 were detected. Isotrip kit of mouse Ig McAbs showed that McAb was IgG1. Culture supernatant and ascitic titres of the antibodies were 1 ： 500, 1 ： 8.0×10^4,when tested by ELISA respectively. Western-blot showed JA9 was bound to 75 ku protein of HHV-6. HHV-6 positive rates were 60% by nested PCR assay and 40% by ELISA in the salivas of oral squamous cell carcinoma patients. Conclusion：The McAbs against HHV-6 might be a useful tool for fundamental studing of HHV-6 and clinic laboratory detection.

关 键 词：人类疱疹病毒6型 单克隆抗体 BALB/C小鼠 

分 类 号：R392.11[医药卫生—免疫学]