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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。
Objective: To investigate the relationship between the activation of pattern recognition receptors (dendritic cells) of dendritic cells (DCs) and the expression of cytokines. METHODS: PPRs and NF-κB signaling pathway-related mRNA expression in mouse bone marrow-derived dendritic cells (BMDC) stimulated with E.coli LLO / OVA were detected by gene chip and RT-PCR Level, and observe the activation status of BMDC and culture supernatant cytokine content. Results: The transient expression of TLR4 mRNA in BMDC was up-regulated at 2 h after stimulation with E.coliLLO / OVA, and the expressions of Myd88, Rip2, Irak1, Irak2, Ikkα, NF-κB1 and NF- The mRNA expression of Icam1, IL-1a, IL-1b, IL-6 and TNF-α was also up-regulated. Card4 (NOD1) mRNA was upregulated in BMDC at 4h after stimulation with E.coliLLO / OVA, The expressions of Rip2, Ikkβ, NF-κB1 and NF-κB2 mRNA were all up-regulated, and the expressions of IFN-γ, TNF-β and CD40mRNA were up-regulated. After 24h stimulation by E.coliLLO / OVA, the expressions of costimulatory molecules and MHC-II molecules in BMDC were up-regulated. The contents of IL-12 and IFN-γ in the culture supernatant were increased. CONCLUSION: Recombinant vaccine E.coli LLO / OVA activates NF-κB signaling through TLR4 and NOD1 receptors and induces mouse BMDC maturation and expression of a series of cytokines, especially IL-12 and IFN-γ.