4株鼠抗人B7-1分子单抗的制备及其生物学活性的鉴定  

Preparation of four monoclonal antibodies against human B7-1 and analysis of their biological characteristics

作　　者：梁文飚[1] 邱玉华[1] 於葛华[1] 戚春建[1] 马泓冰[1] 周璇[1] 王凤鸣[1] 张学光[1] 

机构地区：[1]苏州大学生物技术研究所,苏州215007

出　　处：《中国免疫学杂志》2003年第6期381-384,共4页Chinese Journal of Immunology

基　　金：国家重大基础研究项目资助 ( 2 0 0 1CB5 10 0 3 )

摘　　要：目的 :制备鼠抗人B7 1分子 (mAb)单抗及研究其生物学活性。方法 :以人多发性骨髓瘤细胞转人B7 1基因细胞株XG7 B7为免疫原 ,采用B淋巴细胞杂交瘤技术制备鼠抗人B7 1分子单抗 ;以Westernblot及间接免疫荧光标记法鉴定其特异性和亲和力 ;采用3H TdR掺入实验和Annexin V染色分析测定该单抗的生物学功能。结果 :获得了 4株持续分泌抗人B7 1分子的特异性单抗的杂交瘤细胞株 ,分别命名为 1F11、3H8、6H2和 7B10 ,其分泌的单抗类别分属于小鼠IgG1和IgM ;4株单抗均具有良好的特异性和亲和力 ;1F11、3H8和 6H2能部分阻断B7 1分子基因转导细胞介导的共刺激信号 ,从而抑制T细胞的增殖效应 (抑制率 2 1%～ 5 2 % ) ;且能诱导天然表达B7 1分子的人B系淋巴瘤细胞Raji的凋亡。结论 :成功研制 4株功能性抗人B7 1分子抗体 ,这些抗体在抗异体组织器官移植排斥反应及在B系淋巴瘤的治疗中具有潜在的应用价值。Objective:To prepare the monoclonal antibody(mAb) against human B7 1 and analyse its biological characteristics.Methods:The B lymphocytes hybridization technique was applied by using XG7 B7 cell,a multiple myeloma(MM) cell line transfected with human B7 1 gene,as immunogen;the specificity and the antigen binding activity of mAbs were identified by flow cytometry and Western blot analysis;its biological effects on human PBTC and human B lymphoma cell line were examined by 3H TdR incroporation and annexin V satining.Results:Four mouse anti human B7 1(B7 1) hybridoma(1F11,3H8,6H2,7B10) were obtained.They secrete continuosly and steadily specific anti human B7 1(B7 1) mAb and their subclasses belong to IgG1 and IgM respectively;three of four mAbs could inhibit the proliferation of response cells(the human peripheral blood T lymphocytes),stimulated by costimulatory molecule B7 1.Furthermore,it was found that these mAbs induced the apoptosis of human B lymphoma cell line,Raji,which express naturally human B7 1 molecule,by using annexin V staining analysis after 24 hours of mAb treatment.Conclusion:Sucessefully obtained four mouse anti human B7 1 functional monoclonal antibodies,which have a potential value in anti allogenetic graft rejection and in the therapeutic approach of B lymphoma.

关 键 词：鼠抗人B7—1分子 单抗 制备 生物学活性 鉴定 恶性淋巴瘤 

分 类 号：R392－33[医药卫生—免疫学]