中文摘要：

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。

英文摘要：

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.