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鼠抗人B7 |
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邱玉华 季玉红 郭玲 周照华 王月丹 傅晋翔 张学光
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)、Ra ji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Daud i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。
关键词 B7-1 协同刺激分子 单克隆抗体 恶性淋巴瘤
中国图书分类号 R392.11
Preparation of functional monoclonal antibody against h uman CD80(B7-1) and analysis of its biological effects
QIU Yu-Hua,JI Yu-Hong,GUO Ling et al.
(Institute for Immunology ,Suzhou Medical College,Suzhou 215007)
Abstract Objective:To obtain a monoclonal antibody(mAb) a g ainst human CD80(B7-1) and to study of its biological effects.Method s:The hybridoma cell line was obtained by using the B lymphoma hybrid oma technique after immun ization of Balb/c mice with XG7-B7 the cells.Ascites were induced to produce th e 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 immunofl uorescence.Using counting and trypan blue staining,inhibitory effects of mAb on R aji and Daudi cells were analyzed.The neutralization activity of the 4E 5 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%,respec tived 4E5 can inhibit the growth in Raji and Daudi cells and block the costimula tory 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 basi c studies and find clinical applicat[1] [2] 下一页
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