摘要
目的:探讨树突状细胞(DC)的模式识别受体(PPRs)活化与细胞因子表达的关系。方法:采用基因芯片及RT-PCR检测经E.coliLLO/OVA刺激后小鼠骨髓树突状细胞(bone marrow-derived dendritic cell,BMDC)的PPRs及其下游NF-κB信号通路相关分子mRNA的表达水平,并观察BMDC的活化状况及培养上清液内细胞因子含量。结果:经E.coliLLO/OVA刺激后2h,BMDC出现短暂的TLR4mRNA表达上调,其下游信号途径相关的Myd88、Rip2、Irak1、Irak2、Ikkα、NF-κB1和NF-κB2mRNA均出现表达上调,黏附分子Icam1、细胞因子IL-1a、IL-1b、IL-6和TNF-α的mRNA也表达上调;经E.coliLLO/OVA刺激后4h,BMDC出现Card4(NOD1)mRNA表达上调,其下游信号途径相关的Rip2、Ikkβ、NF-κB1和NF-κB2mRNA均出现表达上调,IFN-γ、TNF-β和CD40mRNA也上调表达。经E.coliLLO/OVA刺激后24h,BMDC表达共刺激分子及MHC-II类分子上调;且培养上清液内IL-12和IFN-γ含量增高。结论:重组疫苗E.coliLLO/OVA经TLR4和NOD1受体激活NF-κB信号通路,诱导了小鼠BMDC成熟并表达一系列细胞因子尤其是IL-12和IFN-γ。
AIM： To investigate the relationship be- tween the activation of pattern recognition receptors and the cytokines expression of dendritic cells. METHODS： After bone marrow-derived dendritic cells （BMDCs） were pulsed by E. coli LLO/OVA, the mRNA expression of pattern recognition receptors and the downstream NF-κB signal path-way associated molecules were detected by microarray hybridization and RT-PCR; the expression of costimulatory molecules, MHC class IT and cytokines were determined by flow cytometry and ELISA. RESULTS. After BMDCs were pulsed by E. coli LLO/OVA for 2 h, the levels of TLR4, Myd88, Rip2, Irak1, Irak2, Ikka, NF-κB1 and NF-KB2 mRNA up-regulated; and the levels of Icaml, IL-1a, IL-1b, IL-6 and TNF-α mRNA up-regulated also; after BMDCs were pulsed by E. coli LLO/OVA for 4 h, the levels of Card4 （Nod1）, Rip2, Ikkl5, NF-κB1 and NF-κB2 mRNA up-regulated, meanwhile, the levels of TNF-γ, TNF-β and CD40 mRNA up-regulated. The expressions of costimulatory molecules and MHC class Ⅱ of the BMDCs up-regulated and the concentration of IL-12 and IFN-γ increased in the supermatant of BMDCs pulsed by E. coli LLO/OVA at 24 h. CONCLUSION： Recombinant E. coli LLO/OVA induces maturation and expression of cytokines, especially IL-12 and IFN-γ, of murine BMDCs through activation of NF-κB signal pathway with TLR4 and NOD1 receptors.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2009年第1期9-12,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
重庆市教委科研基金资助(KJ080319)
关键词
骨髓树突状细胞
重组大肠杆菌
模式识别受体
基因芯片
bone marrow-derived dendritic cell （BMDC）
recombinant E. coil
pattern recognition receptors （PPRs）
microarray
