一株鼠抗人HVEM单克隆抗体的研制及初步鉴定

Preparation and characterization of a mouse anti-human HVEM monoclonal antibody

研制鼠抗人单纯疱疹病毒侵入介质(HVEM)的功能性单克隆抗体,并对其生物学特性进行初步分析。以高表达人HVEM分子的基因转染细胞L929/HVEM作为免疫原,常规免疫小鼠并进行融合,以L929/HVEM作为抗体筛选细胞,L929/mock为对照细胞,筛选出分泌特异性鼠抗人HVEM单克隆抗体的杂交瘤细胞株;采用western blot、间接免疫荧光法和MTT增殖实验对单抗进行生物学特性的分析。结果显示,成功获得一株特异性鼠抗人HVEM的杂交瘤细胞株,命名为2B11。该单抗能够识别肿瘤细胞表达的HVEM分子,且在体外能够显著抑制HVEM介导的T细胞增殖和细胞因子的分泌,从而为进一步研究HVEM介导的免疫学效应提供了物质基础。

To generate a monoclonal antibody（mAb） against human herpesvirus entry mediator （HVEM） and to characterize its biological functions, L929/HVEM, a stable transfectant that expresses human HVEM, was used as the antigen to immunize BALB/c mice. After immunization and fusion, a hybridoma which could specifically secrete anti-human HVEM mAb was selected with L929/HVEM as target cells and L929/mock as the negative control. The specificity of the anti-human HVEM mAb generated by the hybridoma was further identified by testing the binding of the mAb to L929/HVEM transfectant and a group of tumor cell lines using flow cytometry （FCM） and Western blot. Moreover, we investigated its effects on T cell proliferation and cytokine secretion in vitro by MTT assay and ELISA analysis. The results showed that a hybridoma secreting anti-human mAb （named 2Bll） was successfully prepared which could specifically recognize HVEM expressed by the transfectant L929/ HVEM and tumor cell lines and block T cell proliferation as well as cytokine secretion.