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本研究探讨自体调节性T细胞对T细胞淋巴瘤细胞系EL4的增殖抑制作用及其机制。应用免疫磁珠分离法(MACS)分选获得C57BL/6小鼠CD4+CD25+T细胞(Treg细胞),流式细胞术检测分选所得的细胞纯度及Foxp3的表达水平,PT-PCR法检测Foxp3 mRNA的表达,3H-TdR掺入法检测分选的Treg细胞对T细胞淋巴瘤细胞系EL4的体外抑制作用,ELISA方法检测抑制实验中细胞因子TGF-β1和IL-10的水平。结果显示,MACS分选获得CD4+CD25+T细胞纯度达94.52%,Foxp3表达比例为84.72%;分选后的细胞用PT-PCR法可检测到Foxp3mRNA的表达;3H-TdR掺入法证实无论有无抗原呈递细胞(APC)或树突状细胞(DC)存在,自体Treg细胞在体外都能明显抑制EL4细胞的增殖(p<0.05),APC或DC有可能增强这种抑制作用,且单纯DC对EL4细胞的增殖也有抑制作用;实验中可检测到细胞因子TGF-β1和IL-10。结论:自体Treg细胞在体外能够明显抑制淋巴瘤细胞系EL4细胞的增殖,APC或DC可能增强这种抑制作用,且单纯DC对EL4细胞的增殖也有抑制作用。细胞因子TGF-β1和IL-10是Treg细胞发挥抑制作用的途径之一。
This study investigated the autoregulation of T cells on T cell lymphoma cell line EL4 proliferation inhibition and its mechanism. The C57BL / 6 mice CD4 + CD25 + T cells (Treg cells) were sorted by immunomagnetic bead separation (MACS). The purity and Foxp3 expression of the cells were detected by flow cytometry. The expression of Foxp3 mRNA was detected by 3H-TdR incorporation method. The Treg cells were detected by 3H-TdR incorporation in vitro. The levels of TGF-β1 and IL-10 were determined by ELISA. The results showed that the purity of CD4 + CD25 + T cells obtained by MACS sorting was 94.52%, the expression of Foxp3 was 84.72%; the expression of Foxp3 mRNA was detected by PT-PCR after cells were sorted out by 3H-TdR The presence of antigen-presenting cells (APCs) or dendritic cells (DCs) could significantly inhibit the proliferation of EL4 cells in vitro (p <0.05), and the inhibitory effect of APCs or DCs could be enhanced. DC also inhibited the proliferation of EL4 cells; in the experiment, cytokines TGF-β1 and IL-10 could be detected. CONCLUSION: Autologous Treg cells can significantly inhibit the proliferation of EL4 cells in vitro. APC or DC may enhance this inhibition, and DC alone may also inhibit the proliferation of EL4 cells. Cytokines TGF-β1 and IL-10 are one of the pathways by which Treg cells exert inhibitory effects.