CD80人-鼠嵌合抗体对B淋巴瘤细胞株Raji与Daudi的生长抑制及杀伤作用  

Inhibitory and lethal effects of human-mouse chimeric antibody against CD80 on the growth of B lymphoma cell lines Raji and Daudi

作　　者：王艳茹[1] 马泓冰[1] 陈永井[1] 朱华亭[1] 刘玉华[1] 杜阳阳[1] 孙杰[1] 陈明心[1] 邱玉华[1] 

机构地区：[1]苏州大学医学部免疫学系,江苏苏州215123

出　　处：《细胞与分子免疫学杂志》2009年第7期615-618,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省高校高新技术产业发展基金资助项目(JHB05-45);江苏省自然科学基金资助项目(BK2004203)

摘　　要：目的:研究CD80人-鼠嵌合抗体(命名为ch-4E5)对天然高表达CD80分子的人恶性B淋巴瘤细胞株Raji与Daudi的生长抑制及杀伤作用。方法:采用免疫荧光及流式细胞术(FCM)分析ch-4E5对Raji及Daudi细胞膜型CD80分子的识别;将ch-4E5分别加入到Raji及Daudi中共培养,终浓度为10mg/L。在培养的第0、4、10、16、24及48h,采用FCM检测细胞表面Fas及FasL的表达;培养至72h,MTT法检测细胞的增殖;以人外周血PBMC为效应细胞,Raji与Daudi为靶细胞,效靶比为20∶1,加入ch-4E5,终浓度为10mg/L。培养至24h,MTT法分析ADCC效应。结果:ch-4E5与Raji及Daudi的阳性结合率分别为98.6%和96.4%。ch-4E5分别与Raji及Daudi共培养至4h,Raji细胞表面FasL的表达开始上调,16h的阳性表达率为16.8%,与人IgG1对照组1.5%比较明显升高(P<0.01);Fas的表达从10h起逐渐增高,24h达高峰,阳性表达率为15.6%,与人IgG1对照组4.5%比较具有统计学意义(P<0.01)。Daudi的FasL及Fas的变化趋势与Raji一致。ch-4E5分别与Raji及Daudi共培养至72h,细胞增殖的抑制率分别为34.60%和32.64%(P<0.01);ch-4E5介导PBMC对Raji及Daudi的杀伤率分别为55.61%及54.42%(P<0.01)。结论:CD80人-鼠嵌合抗体能够通过其Fab段及Fc段发挥双重的抗肿瘤效应。AIM： To study the inhibitory and lethal effects of human-mouse chimeric antibody against CD80 （named ch-4E5） on the growth of B lymphoma cell lines Daudi and Raji. METHODS： Immunofluorescence and flow cytometry were used to analyze the detection of membrane CD80 in Raji and Daudi by ch-4ES. After the co-culture of ch-4E5 with Raji and Daudi, respectively, at the final concentration of 10 mg/L, the expression of Fas and FasL was observated to be at the 0 h, 4 h, 10 h, 16 h, 24 h and 48 h by direct immunofluorescence and flow cytometry, and the blocking effect on cell growth of ch-4E5 was determined at 72 h by MTT assay. MTT assay was also used to study the ADCC effect with PBMC as effector cells and Raji and Daudi as target cells. The efficiency target ratio was 20： 1. RESULTS： The combination rate between ch-4E5 and Raji and Daudi was 98.6% and 96.4%, respectively. After the coculture of ch-4E5 with Raji and Daudi cells for 4 hs, the expression of FasL in Raji began to up-regulate. It reached the peak at 16 h and its positive rate was 16.8%. Compared with human IgG1 control group, it was increased obviously （P〈0.01）. The expression of Fas increased at 10h, and then reached the top at 2,4 h. The combination rate was 15.6%. There was significant deviation compared with hu- man IgG1 control group （P〈0.01）. Moreover, the expression of FasL and Fas on Daudi was also altered. The trend was similar to these on Raji, and the highest expression was 15.9% and 13.7%, respectively. The inhibitory rate was 34.60% and 32.64% respectively when ch-4E5 with Raji and Daudi had been co-cultured for 72 h （P〈0.01）. Furthermore, ch-4E5 could mediate the ADCC effect and the maximum killing rate was 55.61% and 54.42%, respec-tively（P〈0.01）. CONCLUSION： The human-mouse chimeric antibody against CD80 can inhibit the proliferation of B lymphoma cell lines Daudi and Raji cells through Fab and Fc sections in vitro.

关 键 词：CD80 嵌合抗体 抑制作用 Fas FASL ADCC 

分 类 号：R392.1[医药卫生—免疫学]