重组疫苗E. coli LLO/OVA促进小鼠树突状细胞NOD1受体活化并表达IFN-γ

Recombinant E. coli LLO/OVA promote NOD1 receptor activation and IFN-γ expression of murine bone marrow-derived dendritic cells

目的探讨重组疫苗E.coli LLO/OVA诱导树突状细胞的细胞内受体NOD(nucleotide-binding oligomerizationdomain)活化及表达IFN-γ的作用。方法采用基因芯片杂交分析经E.coliLLO/OVA刺激后小鼠骨髓树突状细胞(bonemarrow-derived dendritic cells,BMDC)Card4/NOD1及其下游NF-κB信号通路相关分子mRNA的表达,RT-PCR检测BMDC的NOD1、NOD2和IFN-γmRNA表达,ELISA测定BMDC培养上清液内IFN-γ含量。结果经E.coli LLO/OVA刺激后4h至8h,BMDC出现Card4/NOD1基因表达上调,其下游信号途径相关分子基因Rip2、Ikkα、Ikkβ、Nfκb1、Nfκb2和IFN-γ均出现表达上调。RT-PCR结果显示该疫苗刺激后BMDC的NOD1与IFN-γ基因表达时段一致。经E.coliLLO/OVA刺激后24h,BMDC培养上清液内IFN-γ含量明显高于E.coli OVA刺激后的BMDC。结论重组疫苗E.coliLLO/OVA诱导小鼠骨髓树突状细胞NOD1受体信号途径活化并促进了IFN-γ表达。

Objective To investigate the role of recombinant E.coli LLO/OVA on the activation of cytosolic receptor nucleotide-binding oligomerization domain(NOD)and IFN-γ expression of murine dendritic cells.Methods After murine bone marrow-derived dendritic cell(BMDC)stimulated by recombinant E.coli LLO/OVA,the mRNA expressions of Card4/NOD1 and downstream NF-κB signal pathway-associated molecules were detected by supperarray hybridization,the expressions of NOD1,NOD2,and IFN-γ were determined by RT-PCR,and IFN-γ con...