联合应用CD3/CD28单克隆抗体对小鼠T淋巴瘤细胞EL4活化的影响

Effect of the combined application of CD3/CD28 monoclonal antibodies on the EL4 T-lymphoma cells activation

目的:研究联合应用CD3/CD28单克隆抗体对小鼠T淋巴瘤细胞EL4活化的影响。方法:ELISA方法检测不同浓度CD3/CD28抗体在不同时间点体外刺激EL4细胞后培养上清中IL-2的分泌;CCK-8方法检测EL4细胞增殖能力;流式细胞术检测EL4细胞周期;RT-PCR检测IL-2、NF-κB和NF-AT转录因子的转录情况。结果:在48小时抗CD3抗体(25μg/ml)与抗CD28抗体(2μg/ml)共刺激EL4细胞明显增加IL-2的分泌和细胞增殖能力,不影响细胞周期。结论:EL4细胞可以作为研究小鼠T淋巴瘤细胞活化的细胞模型。

Objective：To study the effect of the combined application of CD3/CD28 monoclonal antibodies on the activation of EIA cells. Methods： EIA ceils were stimulated by CD3/CD28 antibodies in vitro. The secretion of IL-2 in culture supernatant was de- tected by ELISA assay. EIA cell proliferation ability and cell cycle were analyzed by CCK-8 and flow cytometry methods, respectively. The expression of IL-2 and transcription factors such as NF-KB, NF-AT were analyzed by real-time RT-PCR. Results： The IL-2 secre- tion and EIA cell proliferation ability increased significantly after 48 hours stimulation by CD3 （25 μg/ml） antibody and CD28 （2 μg/ ml） antibody, while the EIA cell cycle was not changed. Conclusion： EIA cell line could act as a cell model of assessing mouse T cell actlvatmn.