【摘要】 目的 :制备鼠抗人B7 1分子的功能性单克隆抗体 ,研究其对高表达相应配基分子细胞的生物学效应。方法 :用两株转人B7 1基因细胞株XG7 B7和L B7分别作为免疫原及检测细胞株 ,利用B淋巴细胞杂交瘤技术制备单克隆抗体。以快速定性试纸分析法鉴定单抗所属的小鼠IgG亚类 ,采用竞争抑制及间接免疫荧光法分析单抗的特异性和亲和力 ,以高表达B7 1分子的恶性淋巴瘤细胞Raji和Daudi为靶细胞 ,分析单抗对其生长的影响。以人多发性骨髓瘤细胞转入B7 1基因细胞株XG1 B7为刺激细胞 ,以人外周血单个核细胞 (PBLs)为反应细胞 ,用MTT法分析单抗的中和活性。结果 :成功地获得了 1株鼠抗人B7 1的功能性单克隆抗体 (克隆 4E5 ) ,属于小鼠IgG1亚类 ,经流式细胞仪分析 ,4E5与PBLs、体外人工诱导的树突状细胞(DCs)、Raji和Daudi的阳性结合率分别为 10 2 %、95 1%、92 7%及 89 2 % ;能完全阻断标准抗人B7 1单抗与相应抗原的结合。同时还发现 ,单抗 4E5能显著地抑制恶性淋巴瘤细胞Raji和Daudi的生长繁殖 ,并能阻断B7分子介导的协同刺激信号的传导。结论 :单克隆抗体 4E5是一株抗人B7 1分子的功能性单抗 ,具有重要的研究和应用价值。更多还原

【Abstract】 Objective:To obtain a monoclonal antibody(mAb) against human CD80(B7 1) and to study of its biological effects.Methods:The hybridoma cell line was obtained by using the B lymphoma hybridoma technique after immunization of Balb/c mice with XG7 B7 the cells.Ascites were induced to produce the mAb.The specificity and affinity of mAb were verified B7 competition and FACS.Expression of B7 1 in PBLs,DCs,Raji and Daudi were studied by indirect immunofluorescence.Using counting and trypan blue staining,inhibitory effects of mAb on Raji and Daudi cells were analyzed.The neutralization activity of the 4E5 determined by MTT assay using PBLs as response cells.Results:The anti CD80(B7 1) was obtained.The expression of B7 1 in PBLs、DC、Raji ad Daudi was 10 2%,95 1%,96 7% and 89 2%,respectived 4E5 can inhibit the growth in Raji and Daudi cells and block the costimulatory signals of B7/CD28.Conclusion:4E5 is a specific and functional anti CD80(B7 1) and has high affinity for its ligand.It may be of significant value in basic studies and find clinical applications.更多还原