【摘要】 目的研究树突状细胞在不同Toll样受体(TLR)配体刺激下的早期反应。方法将鼠骨髓细胞诱导分化得到DC,利用TLR配体刺激,采用芯片分析方法研究其免疫相关基因在9h的表达;采用细胞因子分析系统,检测了12、48h细胞培养上清中细胞因子、趋化因子的产生。结果在给予LPS、PolyI:C、R848、CpG和anti-CD40刺激9h后,IL-1、IL-2rg、IL-13、CCL2、CCL4和CCL7的基因表达明显升高,CCL3、CCL5和CXCL9、CXCL10、CXCL11的表达增加最为显著(P<0.01);而IL-10(PolyI:C除外),CCL-6的表达降低(P<0.05)。培养12h后,IL-1、IL-6、IL-10、IL-12(p40)、IL-12(p70)、TNFa、G-CSF、GM-CSF、KC和MIP-1a(CCL3)产生在5种TLR配体刺激下均显著增加(P<0.01);培养48h后,IL-1、IL-10、IL-12(p40)、IL-12(p70)、TNFa、IFN-γ、G-CSF和MIP-1a(CCL3)产生也同样显著增加(P<0.01)。相对于其它TLR配体,CpG刺激时MIP-1a(CCL3)的产生增加更明显,与基因芯片结果相似。结论本研究揭示了不同TLR配体刺激对DC细胞因子和趋化因子表达量的调节,为TLR配体对DC分化和功能的作用机理提供了更多证据。更多还原

【Abstract】 Objective To determine early responses of DCs to various Toll-like receptor(TLR)ligands.Methods Dendritic cells were prepared from murine bone marrow cells.Using microarray and cytokine array techniques,the expression of immune-related genes were investigated at 9 h after TLR stimulation and the production of cytokines and chemokines was determined at 12 and 48 h.Results Upon exposure to PGN,PolyI:C,LPS,R848,CpG,and anti-CD40 for 9 h,the expressions of IL-1,IL-2rg,IL-13,CCL-2,CCL4,and CCL7 were significantly up-regulated,especially CCL3,CCL5,CXCL9,CXCL10 and CXCL11(P<0.01);whereas IL-10(not in PolyI:C-treated DCs)and CCL-6 were down-regulated(P<0.05).The production of IL-1,IL-6,IL-10,IL-12(p40),IL-12(p70),TNFa,G-CSF,GM-CSF,KC,and MIP-1a(CCL3)were up-regulated respectively in all five different TLR ligands stimulated DCs.Consistent with microarray data,CpG also remarkably promote production of MIP-1a(CCL3).Conclusion Various TLR ligands may rapidly cause DCs to express and produce cytokines and chemokines,which provide clues for understanding the effects of TLRs on the differentiation and function of DCs.更多还原