全人源抗人肿瘤坏死因子相关凋亡诱导配体-R1单克隆抗体诱导肿瘤细胞凋亡的体外实验研究  

A novel human monoclonal antibody to human tumor necrosis factor-related apoptosis-inducing ligand receptor-R1 induces tumor cell apoptosis

作　　者：杨复娇[1] 刘晔[1] 芦凤亮[1] 宗卫娇[1] 梁媛[1] 高翔[1] 金艾顺[1] 

机构地区：[1]哈尔滨医科大学免疫学教研室,150081

出　　处：《国际免疫学杂志》2013年第4期298-302,共5页International Journal of Immunology

基　　金：国家自然科学基金面上项目（81171973）;高校博士学科点专项科研基金（20112307110002）;黑龙江省青年科学基金项目（QC2011C039）

摘　　要：目的探讨本研究组制备的全人源抗人TRAIL—R1单克隆抗体TRAIL—R1-422（TR1-422）诱导肿瘤细胞凋亡的作用。方法利用竞争ELISA法鉴定TR1-422与TRAIL-R1结合特异性；流式细胞分析检测TRl-422与Jurkat细胞表面TRAIL—R1的结合能力；细胞计数法分析TR1-422在不同时间点对Jurkat细胞生存和细胞增殖的影响；镜下观察TR1-422对肿瘤细胞的作用；AnnexinV／PI双染色评价TRl-422诱导Jurkat细胞凋亡的作用。结果TR1-422抗体与重组TRAIL—R1抗原特异性结合；与Jurkat细胞表面TRAIL—R1结合，与无相关IgG抗体组比较荧光强度变化72．3％。1μg/mlTR1-422与10μg／ml抗人IgG交联作用于Jurkat细胞在不同时间点（24h和48h）的生存率分别为72．57％和64．08％，对照组分别为96．14％和95．69％，差异有统计学意义（t=2．534，P〈0．05；t=3．147，P〈0．05）；镜下观察，72h后Jurkat细胞出现大量死亡；24h时AnnexinV阳性率达30％，而TR1-422单独作用为12．8％（t=2．355，P〈0．05）。结论本研究证实制备的全人源抗人TRAIL-R1单抗与抗体交联作用可促进肿瘤细胞凋亡，为抗肿瘤治疗提供基础数据。Objective To evaluate antitumor activity of TR1-422, a novel human monoclonal antibody targeting tumor necrosis factor-related apoptosis-inducing ligand receptor 1 （ TRAIL-R1 ）. Methods The spe- cific binding activity of TR1-422 was detected by competitive ELISA; TRAIL-R1 expression on Jurkat cell＇s was determined by flow cytometric assay; the survival and proliferation of Jurkat cells after treatment with TR1-422 were evaluated at different time point; and Jurkat cell death was observed by microscope. Tumor cell apoptosis induced by TR1-422 was evaluated using Annexin V/PI double staining. Results TR1-422 can bind to recom- binant human TRAIL-R1 specifically and TRAIL-R1 on Jurkat cells but not irrelevanted IgG; the survivals of Jurkat cell was 72.57% and 64.08% 24 h and 48 h after treatment of 1 ug/ml TR1-422 with erosslinker of an- ti-hIgG, whereas the survivals of control groups were 96. 14% ,95.69% respectively （ t = 2. 534, P 〈 0. 05 ; t = 3. 147 ,P 〈 0.05）. We observed a large amount of dead ceils after 3 days under microscope. Furthermore, after 24 h, 30% of the Annexin V positive cells were detected in the group treated by lug/m1 TR1-422 with crosslinker at 24 h, vs 12.8% in the group of 1 ug/ml TRl-422 （ t = 2. 355, P 〈 0.05 ） ; but the H-positive cells weren＇ t detected in each group. Conclusion In the study, we showed that our developed human mono- clonal antibody to human TRAIL-R1, could induce tumor cell apoptosis. It might prompt the development of clinical tumor treatment.

关 键 词：肿瘤坏死因子相关凋亡诱导配体-R1 单克隆抗体 细胞凋亡 

分 类 号：R73-36[医药卫生—肿瘤]