CAJ | 학술논문

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TR1-422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；Annexin V/PI双染色评价TR1-422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/ml TR1-422与10 μg/ml抗人IgG交联作用于Jurkat细胞在不同时间点(24h和48 h)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,72 h后Jurkat细胞出现大量死亡；24 h时Annexin V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.
목적 탐토본연구조제비적전인원항인TRAIL-R1단극륭항체TRAIL-R1-422 (TR1-422)유도종류세포조망적작용.방법 이용경쟁ELISA법감정TR1-422여TRAIL-R1결합특이성；류식세포분석검측TR1-422여Jurkat세포표면TRAIL-R1적결합능력；세포계수법분석TR1-422재불동시간점대Jurkat세포생존화세포증식적영향；경하관찰TR1-422대종류세포적작용；Annexin V/PI쌍염색평개TR1-422유도Jurkat세포조망적작용.결과 TR1-422항체여중조TRAIL-R1항원특이성결합；여Jurkat 세포표면TRAIL-R1결합,여무상관IgG항체조비교형광강도변화72.3％.1μg/ml TR1-422여10 μg/ml항인IgG교련작용우Jurkat세포재불동시간점(24h화48 h)적생존솔분별위72.57％화64.08％,대조조분별위96.14％화95.69％,차이유통계학의의(t=2.534,P＜0.05;t =3.147,P＜0.05)；경하관찰,72 h후Jurkat세포출현대량사망；24 h시Annexin V양성솔체30％,이TR1-422단독작용위12.8％(t=2.355,P＜0.05).결론 본연구증실제비적전인원항인TRAIL-R1단항여항체교련작용가촉진종류세포조망,위항종류치료제공기출수거.
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 specific binding activity of TR1-422 was detected by competitive ELISA ; TRAIL-R1 expression on Jurkat cells 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 recombinant 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 crosslinker of anti-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 cells after 3 days under microscope.Furthermore,after 24 h,30％ of the Annexin V positive cells were detected in the group treated by 1ug/ml TR1-422 with crosslinker at 24 h,vs 12.8％ in the group of 1 ug/ml TR1-422 (t =2.355,P ＜ 0.05) ; but the PI-positive cells weren' t detected in each group.Conclusion In the study,we showed that our developed human monoclonal antibody to human TRAIL-R1,could induce tumor cell apoptosis.It might prompt the development of clinical tumor treatment.