两株鼠抗人VSIG4单克隆抗体的研制及其生物学特性的研究  

Preparation and characterization of two monoclonal antibodies against human VSIG4

作　　者：张世杰[1] 王蕾[2] 汪家敏[1] 陈曦[1] 蒋靓[1] 王月颖[1] 胡玉敏[1] 张学光[1] 顾宗江[1] 

机构地区：[1]苏州大学医学生物技术研究所,苏州215007 [2]苏州大学附属第一医院心内科,苏州215006

出　　处：《中国免疫学杂志》2010年第1期66-69,78,共5页Chinese Journal of Immunology

摘　　要：目的:制备鼠抗人VSIG4分子单克隆抗体(mAb)及对其生物学特性进行鉴定。方法:以高表达膜型VSIG4分子的基因转染细胞L929/VSIG4L为免疫原,常规免疫BALB/c小鼠,采用杂交瘤技术,经流式细胞术检测和多次克隆化培养,筛选出特异分泌鼠抗人VSIG4分子的杂交瘤细胞株;采用间接免疫荧光法、Westernblot、竞争抑制试验及MTT增殖试验等对单抗的生物学特性进行鉴定。结果:获得2株持续、稳定分泌鼠抗人VSIG4mAb的杂交瘤细胞株,分别命名为9A7和9D5。对mAb生物学特性的研究结果表明,2株mAb均能识别巨噬细胞以及Jurkat、THP-1、H446等肿瘤细胞株表面的VSIG4分子。对T细胞增殖抑制的阻断实验显示,这2株抗体对VSIG4分子抑制T细胞增殖均有阻断作用。结论:成功地获得了2株鼠抗人VSIG4功能性mAb,为进一步研究VSIG4及其未知受体的生物学功能奠定了基础。Objective： To prepare anti-VSIG4 manoclonal antibodies and characterize their biological functions. Methods： BALB/c mice were immunized with transfected cell line （L929/VSIG4L） as immunogen. The spleen B cells of the mice were fused with SP2/0 and hybridoma cells were screened with transfected cell line （L929/VSIG4） by FCM. After acquisition of the hybridomas secreting anti-VSIG4 mAb, their biological activities were investigated by indirect immunofluorescence, Western blot, competitive inhibition test, and MTr assay. Results： Two stable hybridomas, 9A7 and 9D5 were obtained, which could continuously secrete specific anti-VSIG4 monoclonal antibodies. The following biological activity studies showed that these monoclonal antibodies could recognize the natural VSIG4 expressed on the macrophages and several cancer cell lines, such as Jurkat, THP-1 and H446. Furthermore, they could block the inhibitory effects of VSIG4 on proliferation of T cells in vitro. Conclusion： Two hybridomas secreting anti-VSIG4 monoclonal antibodies have been established. These monoclonal antibodies provide useful tools for further studying VSIG4＇s biological function and its unknown receptor.

关 键 词：VSIG4 杂交瘤 单克隆抗体 巨噬细胞 

分 类 号：R392.12[医药卫生—免疫学]