核转录因子RelB抑制途径对骨髓树突状细胞表面分子表达的影响研究

The Effect of Nuclear Transcription Factor RelB Suppression on the Surface Molecules of Dendritic Cells

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 explore the effect of nuclear transcription factor RelB suppression on the expression of surface molecules of bone marrow derived dendritic cells and provide a novel pathway for researching tolerogenic dendritic cells. Methods Mouse bone marrow derived dendritic cells were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor （rmGM-CSF） and interleukin-4 （IL-4）,and then purified by CD11c＋ microbead. RelB shRNA lentivirus were prepared by using kits of Invitrogen Corp,and co-cultured with mouse bone marrow dendritic cells. The surface molecules （MHC-II,CD86 and CD40） expression levels were detected by flow cytometry （FCM）. Mature DC （that is,LPS-DC） worked as control group,at the same time,immature DC and LPS RNAi RelB DC were used for comparison. Results The expression levels of co-stimulatory molecules （CD86 and CD40） and MHC-II class molecule were low in DCs of nuclear transcription factor RelB suppression,but significantly high in mature DC （P0.05）. In LPS RNAi RelB DC（that is,after RNAi RelB DC stimulated by LPS） surface the expression levels of CD86,CD40 and MHC-II molecules were low compared with LPS DC（P0.05）. To some extent,the expression levels of surface molecules in RNAi RelB DC were similar to that of immature DC. Conclusion The expression levels of surface molecules were low in DCs with nuclear transcription factor RelB suppression. DCs have the same characteristics with tolerogenic dendritic cells. This is a novel direction to research tolerogenic dendritic cells.