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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。
The monoclonal antibody against murine herpes simplex virus invaded mediator (HVEM) was developed and its biological characteristics were analyzed. The high expression of human HVEM molecule gene transfected cells L929 / HVEM as an immunogen, routine immunization of mice and fusion, L929 / HVEM as an antibody screening cells, L929 / mock as control cells, screened secretory-specific mouse anti-human HVEM monoclonal antibody hybridoma cell lines; Western blot, indirect immunofluorescence and MTT proliferation assay of the biological characteristics of the monoclonal antibody. The results showed that a specific mouse anti-human HVEM hybridoma cell line was successfully obtained and named 2B11. The monoclonal antibody can recognize HVEM molecules expressed by tumor cells and can significantly inhibit HVEM-mediated T cell proliferation and cytokine secretion in vitro, thereby providing a material basis for further study of HVEM-mediated immunological effects.