小鼠骨髓源性树突状细胞和DC2.4中RelB基因表达水平的比较研究  

RelB Expression in Murine Bone Marrow-derived Dendritic Cells and DC2.4

作　　者：郑磊[1] 包杰[1] 王前[1] 顾春瑜[1] 裘宇容[1] 李娟[2] 

机构地区：[1]南方医科大学南方医院检验医学中心,广州510515 [2]南方医科大学南方医院风湿科,广州510515

出　　处：《热带医学杂志》2006年第7期745-748,772,共5页Journal of Tropical Medicine

基　　金：广东省自然科学基金(No.04020404)

摘　　要：目的比较研究体外分离培养的小鼠骨髓来源的树突状细胞(bonemarrow-deriveddendriticcells,BM-DC)和DC2.4中核转录因子RelB基因的表达水平。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联合诱导体系扩增骨髓前体细胞的方法产生未成熟的BM-DC,未成熟的BM-DC在培养结束前18h经LPS刺激获得成熟的BM-DC;DC2.4细胞系用含10%胎牛血清的RPMI1640完全培养液于37℃,5%CO2条件孵箱中培养。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的BM-DC、未成熟的BM-DC和DC2.4,RelBmRNA和其蛋白的表达。结果流式细胞术分析显示MHC-Ⅱ类分子和共刺激分子CD40在未成熟的BM-DC和DC2.4中均呈低水平表达,在成熟的BM-DC中则呈高水平表达;共刺激分子CD86在成熟的BM-DC和DC2.4中均呈高水平表达,而在未成熟的BM-DC中呈低水平表达。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BM-DC和DC2.4中均呈低水平表达;而在成熟的BM-DC中呈高水平表达,成熟的BM-DC与未成熟BM-DC及DC2.4均存在显著差异(P<0.01)。结论RelB基因的表达与小鼠BM-DC的成熟状态密切相关。RelB基因在DC2.4中呈低水平表达,提示DC2.4是一种不成熟的DC细胞系。DC2.4具有的未成熟DC的特点可能与RelB基因的低水平表达有一定关系。抑制骨髓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。Objective To investigate the in vitro expression of avian reticuloendotheliosis viral（v-rel） oncogenerelated B （RelB） in routine bone marrow-derived mature dendritic cell,immature BM-DC and DC2.4. Methods Bone marrow was collected from femur and tibias of C57BL/6 mice in sterile condition, bone marrow ceils were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor （rmGM-CSF） and interleukin-4 （rmIL-4）. The immature BM-DCs were induced to differentiate into mature BM-DCs by LPS for the last 18 h in the culture. DC2.4 ceil line was cultured in RPMI 1640 complete medium containing 10% FBS in 37℃, 5% CO2 incubator. DCs＇ phenotype was detected by flow cytometry （FCM）. RelB mRNA and protein were detected in the mature BM-DC, immature BM-DC and DC2.4 by RT-PCR and immunofluorescence staining, respectively. Results CD40, CD86 and MHC-Ⅱ were expressed in mature BM-DC at high levels but at low leveles in immature BM-DC and DC2.4 ceil line. The results of RT-PCR and immunofluorescence staining both demonstrated that the expression level of RelB was low in immature BM-DC and DC2.4 but high in the mature BM-DC. The differences in the expression levels of CD40, DC86, MHC-Ⅱ and ReIB between the mature BM-DC and immature BM-DC or DC2.4 were significant （P〈0.01）. Conclusion RelB expression is closely associated with the maturity of BM-DCs. RelB was expressed in DC2.4 at a low level, which indicated that DC2.4 is a immature dendritic cell line. The immature characteristics of DC2.4 may be related with low expression level of RelB. Suppress the expression of RelB may lead to the formation of tolerogenic DCs.

关 键 词：小鼠 骨髓源性树突状细胞 DC2.4 RELB 基因表达 

分 类 号：R392.12[医药卫生—免疫学]