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目的探讨核转录因子RelB抑制途径对小鼠骨髓树突状细胞(bone marrow dendritic cells)的表面分子表达的影响,为致耐受树突状细胞的研究提供新方法。方法 rmGM-CSF和rmIL-4联合诱导培养体系培养小鼠骨髓树突状细胞,免疫磁珠方法纯化;用慢病毒载体制备RelB shRNA慢病毒,与小鼠骨髓树突状细胞共培养,流式细胞术观察树突状细胞表面分子MHC-II、CD86和CD40的表达,设LPS-DC对照组、未处理组和LPSRNAi RelB DC组。结果核转录因子RelB抑制的树突状细胞表面分子MHC-II、CD86和CD40均低水平表达,显著低于成熟DC表面分子的表达(P<0.05),且经LPS刺激后(LPS RNAi RelB DC)DC表面上述三类分子的表达水平仍显著低于LPS-DC组(P<0.05),与未处理组(immature DC)表面分子表达水平相当。结论核转录因子RelB抑制的骨髓树突状细胞表面分子表达水平低,呈现出致耐受的树突状细胞的特点,是致耐受树突状细胞研究的一种新方法。
Objective To investigate the effect of RelB repression pathway on the surface molecule expression of mouse bone marrow dendritic cells, and to provide a new method for the study of tolerogenic dendritic cells. Methods Murine bone marrow dendritic cells (DCs) were cultured with rmGM-CSF and rmIL-4, and purified by immunomagnetic beads. RelB shRNA lentivirus was prepared by lentiviral vector and co-cultured with mouse bone marrow dendritic cells Cell surface expression of dendritic cell surface molecules MHC-II, CD86 and CD40, LPS-DC control group, untreated group and LPSRNAi RelB DC group. Results The expression of MHC-II, CD86 and CD40 on dendritic cells inhibited by RelB was significantly lower than that of mature DCs (P <0.05), and LPS RNAi RelB DC ) DCs expression levels of these three types of molecules were still significantly lower than the LPS-DC group (P <0.05), and the surface molecules of untreated group (immature DC) expression level is equivalent. Conclusions The expression of dendritic cells, which is induced by nuclear transcription factor RelB, is low on the surface of dendritic cells and shows the characteristics of tolerogenic dendritic cells. Therefore, it is a new method to induce dendritic cells.