摘要
目的研究可溶性的抗CD47单抗(B6H12)对树突状细胞(DC)的免疫调控作用及分子机理。方法分离人外周血单核细胞,联合应用rhGM-CSF、IL-4、细菌脂多糖(LPS)在体外诱导扩增DC,并在培养体系中添加抗CD47单抗(B6H12)。采用透射电镜观察DC形态,流式细胞仪检测DC膜表面分子,ELISA定量检测DC释放IL-12P70水平,Brdu-ELISA法检测DC刺激同种异型淋巴细胞增殖,凝胶电泳迁移率改变实验(EMSA)检测NF-κB活性。结果(1)经抗CD47单抗(B6H12)处理的DC,膜表面CD80、CD86、CD1a、CD83、DR表达显著降低(P<005);其释放IL-12P70的水平及刺激同种异型淋巴细胞增殖的能力也显著低于对照组(P<001)。(2)B6H12单抗处理DC组其NF-κB活性显著降低(P<005),且该效应呈剂量依赖性。结论抗CD47单抗可通过抑制NF-κB活性而影响DC的发育、成熟及功能。
AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-κB activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P<0.05). The data of mixed leukocyte reaction and IL-12P70 production were consistent with the flow cytometry results (P<0.01). Nuclear extracts from the anti-CD47 mAb-treated DCs revealed a decrease in NF-κB binding activity. CONCLUSION: The anti-CD47 mAb exerts a negative effect on the maturation and function of in vitro cultured DCs via inhibiting of NF-κB binding activity. [
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第12期2248-2252,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30270571)
教育部博士点专项基金项目(No.20020335080)
