小鼠骨髓成熟与不成熟树突状细胞中RelB基因的表达

目的:探讨体外分离培养的小鼠骨髓来源的成熟与未成熟DC中核转录因子RelB(avianreticuloendotheliosisviral(v-rel)oncogenerelatedB)基因的表达。方法:无菌从C57BL/6小鼠股骨和胫骨中取出骨髓细胞,利用rmGM-CSF和rmIL-4联合诱导骨髓前体细胞产生未成熟的DC,未成熟的DC在培养结束前18h经LPS刺激获得成熟的DC,用流式细胞术分析它们的表型,用RT-PCR和免疫荧光染色法检测成熟与未成熟DC中,RelBmRNA和其蛋白的表达。结果:流式细胞术分析显示未成熟的DC中MHC-Ⅱ类分子和共刺激分子(CD86和CD40)呈低水平表达;而成熟的DC则呈高水平表达。RT-PCR和免疫荧光染色法检测结果均显示,RelB基因在未成熟的DC中呈低水平表达;而在成熟的DC中呈高水平表达,两者比较具有统计学意义(P<0.01)。结论:RelB基因的表达与小鼠骨髓来源的DC的成熟状态密切相关。抑制DC中RelB基因的表达,有可能诱导产生具有耐受原性的未成熟的DC。

RelB gene expression in murine bone marrow-derived mature and immature dendritic cells

AIM: To explore the expression of avian reticuloendotheliosis viral(v-rel) oncogene-related B (RelB) mRNA in vitro in murine mature and immature myeloid dendritic cells (DCs). METHODS: The bone marrow was collected from the femur and tibias of C57BL/6 mice in sterile condition. Bone marrow precursors were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and interleukin-4 (rmIL-4) to produce immature DCs. Immature DCs were stimulated by LPS 18 h before the end of culture to become mature. DCs' phenotype was detected by flow cytometry (FCM). RelB expression and protein level were determined by RT-PCR and immunofluorescence staining. RESULTS: The expression level of co-stimulatory molecules (CD86 and CD40) and MHC-II class molecule were low in immature DCs, whereas high in mature DCs. RelB expression was significantly higher in mature DCs than in immature DCs (P<0.01). CONCLUSION: RelB expression is closely associated with the maturative situation of DCs. Inhibition of RelB expression in DCs may induce tolerogenic DCs.