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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。
Objective: To analyze the dynamic changes of T cell proliferation in mouse mixed lymphocyte reaction (MLR) by flow cytometry (FCM). Methods: The dendritic cells (DCs) stimulated by lipopolysaccharide (LPS) and normal saline (NS) were respectively challenged with H22 tumor cell antigen peptides and then incubated with mouse T cells (TC) Allo-MLR were performed as LPS + DC + TC group and NS + DC + TC group, respectively. LPS + TC group and blank TC control group were also established. TC cultured in vitro, TC, FCM were used to detect the cell cycle of TC at 0d, 1d, 2d, 3d, 4d, 5d and 6d respectively. Results: S phase cells and cell proliferation index in LPS + DC + TC group and NS + DC + TC group first increased and then decreased with the prolongation of culture time, but decreased in LPS + DC + TC group and blank TC control group (P < 0.05). The difference between the two groups was statistically significant (P <0.05). Conclusion: FCM can objectively and accurately reflect the dynamic changes of lymphocyte proliferation from single cell level.