CFSE示踪与流式细胞仪检测法研究环磷酰胺对T淋巴细胞增殖的影响

目的利用定量分析羧基荧光素乙酰乙酸(carboxyflu-orescein diacetate succinimidyl ester,CFSE)时间系列数据的方法研究淋巴细胞细胞增殖动力学及免疫抑制剂对淋巴细胞增殖抑制机制的研究。方法环磷酰胺造成小鼠免疫抑制模型,并用CFSE染料给对照组和给药组动物的淋巴细胞染色,通过数学模型拟合法分析数据。结果对照组和给药组淋巴细胞增殖进入第一代分裂的时间分别是27.17h和22.88h,每代细胞死亡率分别为20%和40%;进入分裂前的细胞半衰期分别为31.53h和43.32h。结论通过数学拟合法对CFSE数据进行定量分析,探讨药物对淋巴细胞增殖影响的机制,该实验中的药物环磷酰胺抑制淋巴细胞增殖的方式可能是使细胞增殖时每代细胞的死亡率增加所致。

The research of lymphocyte proliferation using CFSE and FCM

Aim To quantitatively analyze the lymphocyte proliferation using the time-series data of carboxyfluorescein diacetate succinimidyl ester(CFSE).Methods The model of immune suppression in mice was created by cyclophosphamide and CFSE dye was used for staining the lymphocyte of both control group and treatment group animals. The data were analyzed through mathematical model-fitting.Results The first generation of cell proliferation of the control group and treatment group were 27.17 h and 22.88 h; cell death rates in each division were respectively 20% and 40%; the half-life of cells before the proliferation was respectively 31.53 h and 43.32 h.Conclusions The mathematical fitting of CFSE data can quantitatively analyze the mechanism of the drugs attecting the proliferation of lymphocyte.In this experiment the mechanism of cyclophosphamide inhibiting lymphocyte proliferation may be due to the increase of cell death rate of each division caused by cyclophosphamide.