活体染料CFDA-SE在淋巴细胞增殖研究中的应用

Application of vital dye CFDA-SE to study lymphocytic proliferation

目的 :探讨活体染料CFDA SE在淋巴细胞增殖研究中的应用价值。方法 :利用CFDA SE染色、荧光抗体标记和流式细胞术 ,检测淋巴细胞及其亚群在多克隆刺激剂作用下荧光强度的变化 ,并应用相关软件分析其增殖情况。结果 :PDB +ion omycin或ConA刺激 4 8h后均出现淋巴细胞分裂 ,表现为CFSE荧光强度的系列减半。环孢菌素A(CsA)可抑制ConA促进淋巴细胞增殖的作用 ,CFSE荧光无减半。ConA刺激4 8h后 ,出现CD4 + T细胞和CD8+ T细胞增殖不同步的现象 ,72h后这一现象更加明显。经ModFitTM 软件拟合后 ,得到的各项增殖指标显示 ,ConA对CD8+ T细胞的促增殖作用强于对CD4 + T细胞的作用。结论 :CFDA SE染色结合荧光抗体标记和流式细胞术 。

AIM: To explore the application value of vital dye CFDA SE to study of lymphocytic proliferation. METHODS: CFDA SE staining, fluorescence antibody labeling, flow cytometry and related software were used to detect the fluorescence intensity and analysis the proliferation kinetics of lymphocytes and their subsets after stimulation with polyclonal stimulators. RESULTS: Lymphocytes divided after stimulation of PDB+ionomycin or ConA for 48 h, manifesting the serial halving of fluorescence intensity. CsA inhibited the proliferative effect of ConA on lymphocytes and no CFSE fluorescence halving were seen. Proliferation of CD4 +T cells and CD8 + T cells were asynchronous after ConA stimulation for 48 h, which became more obvious at the time of 72 h. Proliferation related indexs got ten from ModFit TM software showed that the proliferative effect of ConA on CD8 + T cells was stronger than that on CD4 + T cells. CONCLUSION: CFDA SE staining combined with fluorescent antibody labeling and cytometry were powerful tools for analysis of lymphocytic proliferation.