抗人CD86嵌合抗体对B淋巴瘤细胞株Raji生长抑制及杀伤作用研究  

Study on suppression and lethal effect of anti-human CD86 chimeric antibody to a human malignant B lymphoma cell line Raji

作　　者：刘玉华[1] 孙杰[1] 胡玲玲[1] 王艳茹[1] 马鸿冰[1] 程钢[1] 徐耀瑜[1] 杜阳阳[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,苏州215123

出　　处：《中国免疫学杂志》2009年第6期513-516,共4页Chinese Journal of Immunology

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

摘　　要：目的:研究CD86嵌合抗体ch1D1对天然高表达CD86分子的人恶性B淋巴瘤细胞株Raji增殖抑制及杀伤效应。方法:经流式细胞术检测ch1D1对Raji细胞表面CD86分子的识别;经ch1D1(终浓度10μg/ml)处理Raji细胞,MTT法检测对Raji细胞增殖的抑制效应;流式细胞术检测ch1D1诱导Raji细胞表面FasL及Fas的表达;MTT法检测ch1D1介导的外周血单个核细胞(PBMCs)对Raji为靶细胞的ADCC效应。结果:ch1D1可特异性识别Raji细胞表面CD86分子,阳性结合率为97.5%。ch1D1对Raji细胞增殖的抑制率为27.1%,较鼠源性亲本抗体1D1组(抑制率为38.8%)略下降,但两者统计学比较无差异(P>0.05)。与CD80嵌合抗体ch4E5联合应用抑制率可达56.3%,明显高于ch1D1单独作用的抑制率(P<0.05)。ch1D1作用4小时后,Raji细胞表达FasL水平开始升高,处理72小时阳性表达率为6.8%,较亲本抗体1D1组(阳性表达率3.2%)升高,但统计学比较无显著差异(P>0.05);经ch1D1作用24小时,Raji细胞Fas表达水平开始上调,阳性率为6.1%,较1D1组(阳性表达率4.8%)升高,但无统计学差异(P>0.05)。与ch4E5联合应用均较ch1D1单独作用Fas及FasL的表达水平升高,但统计学比较仍无显著差异(P>0.05)。经ch1D1处理4、24、48、72小时,介导PBMC对Raji细胞杀伤率分别为25.0%、52.0%、71.3%、72.7%,与人IgG1及鼠源亲本抗体对照组相比均明显升高,具有显著差异(P<0.05)。结论:抗人CD86嵌合抗体可抑制高表达CD86分子的肿瘤细胞增殖并诱导其凋亡,与嵌合抗体Fc段介导的ADCC效应具有协同抗肿瘤作用。Objective：To investigate the suppression and lethal effect of chimeric antibody ch1D1 to Raji cells, a human malignant B lymphoma cell line, which has the property of naturally high expression of CD86 molecule. Methods：The recognition of ch1D1 to Raji cell surface molecule CD86, under the condition of co-culture of chtD1 and Raji with a final concentration of ch1D1 at 10ug/ml was examined by flow cytometry. The inhibitory effect of chl D1 on growth of Raji cells was assessed by MTT assay. Expression of FasL and Fas on Raji cells induced by ch1D1 antibody was detected by flow cytometry. For analysis of the ADCC effect of PBMCs on Raji cells, after treatment with ch1D1 for 4 h, 24 h,48 h,and 72 h,cytotoxity of peripheral blood monocytes （PBMCs） （used as effector cells） to Raji （as target cells together） was detected via MTT assay. Results： Ch1D1 could specifically recognize surface molecule CD86 on Raji cell with a positive percentage of 97.5 %. Ch1D1 alone could suppress the propagation of Raji cells in vitro, with the inhibition rate of 27.1%, with decrease when compared to the parental antibody 1D1 （38.8 % ）, but there was no significant deviation between the groups by statistical analysis （P 〉 0.05）, when ch1D1 reacted in combination with anti-CD80 human-mouse chimeric antibody ch4ES, the inhibition rate rised to about 56.3 %, with apparently augmentation when compared with that of chl D1 alone（ P 〈0. 05）. Raji cell surface molecule, FasL, began to express at d h after the action of chl D1 and its expression got increase as the time past, its positive rate was up to 6.8 % at 72 h, significant increase compared to that of the parental antibody 1 D1 group （3.2 % ）, and there was no significant deviation compared with 1 D1 by statistics comparision（ P 〉 0. 05）;The expression of Fas began to increase up to 6.1% at 24 h after the action of chl D1, with increase when compared that of to the 1 D1 group （4.8 % ） with no significant deviation compared with 1 D1 by statisti

关 键 词：CD86 嵌合抗体 淋巴瘤 凋亡 ADCC效应 

分 类 号：R392.11[医药卫生—免疫学]