一株鼠抗人4-1BBL功能性单克隆抗体的研制及其生物学特性的研究  

Preparation of a functional monoclonal antibody against human 4-1BBL molecule and analysis of its biological characteristics

作　　者：王旭东[1] 周斌[2] 李文香[1] 於葛华[1] 张光波[2] 张学光[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临床免疫学实验室,苏州215006

出　　处：《中国免疫学杂志》2008年第5期439-442,451,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金青年科学基金项目(30400394);国防科工委重点项目(A3820060130)资助

摘　　要：目的:鼠抗人4-1BBL功能性单克隆抗体的研制及其生物学特性的鉴定。方法:以高表达人4-1BBL分子的基因转染细胞L929/4-1BBL为免疫原,常规免疫BALB/c小鼠,采用B淋巴瘤杂交瘤技术进行细胞融合,经流式细胞术分析和多次克隆化培养,筛选特异性杂交瘤细胞株;采用Ig亚型快速定性试纸法、染色体核型分析、竞争结合抑制试验、间接免疫荧光法对单抗的生物学特性进行鉴定,并采用体外细胞计数和ELISA检测分析单抗对单核细胞的作用。结果:获得1株持续、稳定分泌鼠抗人4-1BBL单抗的杂交瘤细胞株,命名为3E7。该单抗能识别人白血病细胞株和单核细胞表面的4-1BBL分子。对单抗生物学功能的研究结果表明,该单抗能有效地促进外周血单核细胞体外生长,并促进IL-6和TNF-α的分泌。结论:成功地获得了一株鼠抗人4-1BBL功能性单克隆抗体3E7,该单抗能特异地识别人4-1BBL分子,并有效地通过激发4-1BBL逆向信号通路促进单核细胞的体外生长及细胞因子的分泌。Objective： To prepare functional monoclonal antibodies against human 4-1BBL molecule and analysis of their biological characteristics.Methods： Female B ALB/c mice of 6-8 weeks old were immunized with 4-1BBL transfectant （L929/4-1BBL） as immunogen. The spleen B cells of the mice were fused with sp2/0 and hybridoma cells were screened with 4-1BBL transfectant （L929/4-1BBL） by FCM. The biological characteristics of antibody were investigated by rapid isotyping analysis, karyotype analysis, competitive inhibition test etc. Furthermore, the growth of monocytes in vitro was determined by cell number counting and the cytokine concentration in the supernatants was assayed by ELISA. Results- One hybridoma cell line named 3E7 was obtained, which had the property of secreting anti-human 4-1BBL monoclonal antibody continuously and steadily. This mAb specifically recognized human 4-1BBL molecules. The experimental results manifested that mAb 3E7 could effectively enhance the growth of monocytes and the high level IL-6 and TNF-α secretion. Conclusion： One hybridoma cell line which can secret a functional mouse anti-human 4-1BBL mAb has been developed successfully. This mAb can specifically recognize human 4-1BBL and regulate growth and functions of monocytes in vitro.

关 键 词：4-1BBL 单克隆抗体 共刺激信号 杂交瘤 

分 类 号：R392.12[医药卫生—免疫学]