BrdU掺入与T细胞的活化和细胞因子表达的关系  

Correlation of BrdU incorporation with activation and cytokine expression of T cells

作　　者：李丽[1] 池沛冬[1] 孟锐[1] 杨滨燕[1] 吴长有[1] 

机构地区：[1]中山大学基础医学院免疫学教研室,热带传染病教育部重点实验室,广东广州510080

出　　处：《细胞与分子免疫学杂志》2008年第1期72-75,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：广东省自然科学基金团队项目(05200176);广州市科技计划科技攻关引导项目科技交流合作专项国家、省市联动项目(2005Z3-C7461,2006Z3-C7211)

摘　　要：目的:探讨利用流式细胞术(FCM)检测细胞增殖(BrdU掺入)与T细胞的活化及细胞因子表达的关系。方法:正常人PBMC经PMA+Ionomycin刺激不同时间,在培养结束前1 h加入BrdU,利用抗BrdU以及多种抗细胞表面和抗细胞因子抗体标记,FCM检测。结果:体外刺激PBMC 48 h后,可见T细胞有BrdU掺入,但随着时间的延长,掺入率没有明显增加。对比分析BrdU掺入与活化分子表达的关系表明,CD69在刺激后8 h达高峰,而CD25则需要24 h后达到高峰。另外,BrdU掺入与细胞因子表达没有明显关系,当PBMC刺激后8 h,即有大量IFN-γ的表达,而当培养时间延长到24、48和72 h后,IFN-γ的表达没有明显改变。OKT3+antiCD28刺激T细胞后BrdU的阳性率高于PMA+Ionomycin刺激的T细胞,CD8+T细胞掺入率高于CD4+T细胞。结论:利用FCM检测BrdU+细胞可以用于评价T细胞的增殖反应,但在多克隆刺激的条件下,只有少数细胞发生增殖,并且BrdU掺入率受刺激剂种类和时间的影响,BrdU掺入与活化分子和细胞因子的表达均没有明显关系。AIM： To investigate the correlation of BrdU incorporation with the activation and cytokine expression of T cells. METHODS： PBMCs from healthy persons were isolated and stimulated by PMA plus Ionomycin at different periods of time, BrdU was then added to the cells one hour before the end of culture. The cells were harvested and stained with anti-BrdU, anti-cell surface and intracellular antibodies. Then the cells were washed and analyzed by flow cytometer. RESULTS： The peak of BrdU incorporation was observed in T cells after they were stimulated for 48 hours in vitro, but no further increase of the peak of BrdU incorporation was found after incubated for a longer period of time. The comparison made between BrdU incorporation and cell activation indicated CD69 expression reached the peak after stimulated for 8 hours whereas CD25 was at the peak after stimulated for 24 hours. Furthermore, no correlation between BrdU incorporation and cytokine production was observed. High frequency of IFN-γ producing cells was detected after stimulated for 8 hours but no obvious increase was observed for a longer period of time. When PBMC were stimulated with OKT3 plus antiCD28, the percentage of BrdU ＋ T cells was higher than that stimulated by PMA plus Ionomycin. Similarly, the percentage of BrdU ＋ CD8 ＋ T cells was higher than that of BrdU ＋ CD4＋ T cells. CONCLUSION： The percentage of BrdU ＋ cells can be detected by flow cytometer to evaluate the proliferation of T cells. Only a few T cells proliferate after polyclonal stimulation and BrdU incorporation is dependent on stuimulants and time of stimulation. Therefore, BrdU incorporation is not correlated with activation markers and cytokine production.

关 键 词：BRDU 增殖 流式细胞仪 细胞活化 

分 类 号：R392[医药卫生—免疫学]