三种方式活化T细胞CD69分子表达观察  

The Influence of three different activators on CD69 expression of CD3＋ lymphocytes

作　　者：苑振新 

机构地区：[1]泰安市肿瘤防治院检验科,271000

出　　处：《国际免疫学杂志》2015年第6期522-524,共3页International Journal of Immunology

基　　金：基金项目：山东省泰安市科技发展计划（20074043）;山东省泰安市科技发展计划（201440774-38B）

摘　　要：目的观察CD3单克隆抗体（CD3mAb）、佛波醇酯＋离子霉素（PMA＋IM）、结核杆菌低分子多肽抗原（Mtb-Ag）活化CD3＋T细胞CD69分子的表达情况。方法分离获取健康人外周血单个核细胞（PBMC），用CD3mAb（A组）、PMA＋IM（B组）、Mtb．Ag（C组）刺激PBMC，采用流式细胞术检测。CD3＋T细胞和∞T细胞0、6、12、24、48和72hCD69分子的动态表达情况。结果CD3＋T细胞CD69分子的动态表达情况：CD3mAb刺激组24h达高峰，为56．2％；PMA＋IM刺激组6h达高峰，为99．5％；Mtb-Ag刺激组24h达高峰，为16％。各组比较F=322．528，P〈0．001；两两比较，均P〈0．001。γδT细胞CD69分子的动态表达情况：CD3mAb刺激组24h达高峰，为55．1％；PMA＋IM刺激组6h达高峰，为99．3％；Mtb-Ag刺激组24h达高峰，为75．2％。各组比较F=21．170，P〈0．001；两两比较：CD3mAb与PMA＋IM组比较，PMA＋IM组与Mtb—Ag组比较，均P〈0．001；Mtb-Ag与CD3mAb比较P=0．646。结论PMA＋IM的刺激，CD3＋T细胞和γδT细胞活化达高峰的用时比CD3mAb、Mtb-Ag的刺激要短，CD69的表达率也比CD3mAb、Mtb-Ag的刺激要高，应作为三种方法中CD3＋T细胞活化的首选方法。Objectives To study the influence of three different activators on CD69 expression of CD3 ＋lymphocytes. Methods Peripheral blood mononuclear cells （PBMCs） were separated from the blood of healthy contributors and stimulated with CD3 monoclonal antibodies （CD3mAb,group A） ,phorbol 12-myristate 13-acetate plus ionomycin （ PMA ＋ IM, group B）, or low molecular peptide antigen of myeobacterium tuberculo- sis （Mtb-Ag,group C）. The dynamic expression of CD69 molecule on CD3 ＋ T cells and γδ cells was detec- ted by flow cytometric analysis at 0,6,12,24,48, and 72 hours after stimulation. Results The level of CD69 expression on CD3 ＋T cells in group A reaches the summit of 56.2% at 24 h. It reaches the summit of 99.5% at 6 h in group B and 16% at 24 h in group C respectively （ F = 322. 528, P 〈 0. 001, pairwise comparison ： P 〈 0. 001 ）. The level of CD69 expression on γδ cells in group A reaches the summit of 55.1% at 24 h. It rea- ches the summit of 99.9% at 6 h in group B and 75.2% at 24h in group C separately （ F = 21. 170, P 〈 0. 001, pairwise comparison ：P 〈 0.001 when A was compared with B and B compared with C,P = 0. 646 when C was compared with A）. Conclusions Among the three activators, PMA ＋ IM is most efficient in activating CD3＋ T cells and -/GT cells. It can induce the highest level of CD69 expression in the shortest time period.

关 键 词：CD3抗体 佛波醇酯 离子霉素 结核杆菌低分子多肽抗原 CD3+T细胞 CD69分子 

分 类 号：R446.6[医药卫生—诊断学]