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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γδT细胞0、6、12、24、48和72 h CD69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:CD3 mAb刺激组24 h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 mycobacterium tuberculosis (Mtb-Ag,group C).The dynamic expression of CD69 molecule on CD3 + T cells and γδT cells was detected 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 γδT cells in group A reaches the summit of 55.1％ at 24 h.It reaches 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 γδT cells.It can induce the highest level of CD69 expression in the shortest time period.