应用流式细胞术动态观测小鼠树突状细胞诱导的T细胞增殖的变化

Employing FCM to observe the dynamic changes of T lymphocyte proliferation stimulated by dendritic cells in mice

目的:运用流式细胞术(FCM)分析小鼠混合淋巴细胞反应(MLR)中T细胞增殖的动态变化。方法:把分别经脂多糖(LPS)和生理盐水(NS)刺激的小鼠树突状细胞(DC)经H22肿瘤细胞抗原肽冲击后,以1:50的比例与小鼠T细胞(TC)进行同种MLR,分别作为LPS+DC+TC组和NS+DC+TC组,同时设LPS+TC组和空白TC对照组。经体外培养,分别于反应的第0d、1d、2d、3d、4d、5d、6d收集TC,FCM检测TC细胞周期。结果:随培养时间的延长,LPS+DC+TC组、NS+DC+TC组S期细胞及细胞增殖指数均先增后减、LPS+TC组和空白TC对照组则呈下降趋势(P<0.05),各组间比较差异有统计学意义(P<0.05)。结论:FCM可以从单细胞水平客观、准确地反映淋巴细胞增殖的动态变化。

Aim： To observe the dynamic changes of T lymphocyte proliferation after mixed lymphocyte reaction （MLR） using flow cytometry （ FCM ）. Methods ： Mouse dendritic cells（ DC ） were induced by lipopolysaccharide （LPS） or by NS in vitro, and then plused ex vive with hepatocellular carcinoma H22 antigen. Then the above DC were mixed with mouse spleen T lymphocytes （TC） by 1 ： 50. There were 4 groups in total, namely LPS ＋ DC ＋ TC group, NS ＋ DC ＋ TC group, LPS ＋ TC group and blank control group. On the 1 st,2nd ,3rd ,4th,5th ,6th d after MLR, TC were collected to detect the cell cycle using FCM. Results：TC at S phase and cell proliferation index both increased at first stage and then decreased afterwards in LPS ＋ DC ＋ TC group and NS ＋ DC ＋ TC group, while these parameters had a decreasing tendency in LPS ＋ TC group and blank control group. Conclusion ： The consequence of FCM can objectively and exactly represent the dynamic changes of TC on the single cell level.