白内障术后后囊浑浊的防治和抗人晶状体上皮细胞单克隆抗体的制备  

The study of prevention of posterior capsular opacification:Preparation and identification of monoclonal antibody specific to human lens epithelial cell

作　　者：孙靖[1] 李筱荣[1] 孙慧敏[1] 袁佳琴[1] 

机构地区：[1]天津医科大学眼科中心,天津300070

出　　处：《眼外伤职业眼病杂志》2007年第3期168-171,共4页Journal of Injuries and Occupational Diseases of the Eye with Ophthalmic Surgeries

基　　金：天津市21世纪青年科学基金(003702711)

摘　　要：目的制备可特异性识别人晶状体上皮细胞的单克隆抗体,筛选特异性强亲和力高的克隆。方法以原代培养人晶状体上皮细胞作为免疫原,使用杂交瘤技术制备抗人晶状体上皮细胞单克隆抗体(monoclonal antibody,McAb)。小鼠腹腔接种杂交瘤细胞制备腹水,用免疫荧光、免疫组化法对其特性进行鉴定。结果获得了1株稳定分泌抗人晶状体上皮细胞McAb的杂交瘤细胞系,其亚类为IgM。免疫荧光和免疫组化检测结果显示,此McAb与人晶状体上皮细胞膜上的抗原反应,与人眼其他组织呈阴性反应。结论所获抗体可特异性识别人晶状体上皮细胞,可进一步制备免疫导向药物,用于白内障术后后囊浑浊的治疗。Objective To prepare and identify high-affinity monoclonal antibody （McAb） specific to human lens epithelial cell, and preliminarily study its characteristics. Methods BALB/ c mice were immunized by intrasplenic injection of cultured human lens epithelial cells. The immunized mouse spleen cells were fused with myeloma cell line NS-1. The McAb against human lens epithelial cell was screened by indirect immunofluorescence techniques. The large quantities ofMcAb were prepared by the hyhridoma cells inoculated to the peritoneal cavity of BALB/c mouse. Its specificity was confirmed both by immunohistochemical analysis and＇ by indirect immunofluorescence,techniques. Results One hyhridoma cell line secreting anti - human lens epithelial cell McAh was successfully obtained, its subclass is IgM. Both immunohistochemical and immunofluorescence examinations demonstrated that the McAb was only positive to human lens epithelia! cell membrane and negative to the other tissues of human eyeball. Conclusions The McAh against human lens epithelial cell was-successfully prepared, which can serve as a carder of the target - directing agents designated to actively identify and selectively kill human lens epithelial cells. The agents may be more efficient in prevention and treatment of posterior capsular opacification （PCO）.

关 键 词：晶状体上皮细胞 单克隆抗体 杂交瘤细胞 后囊浑浊 

分 类 号：R779.66[医药卫生—眼科]