OVA经TLR4和NOD1受体诱导树突状细胞表达细胞因子.doc

LLO/OVA经TLR4和NOD1受体诱导树突状细胞表达细胞因子
(作者:___________单位: ___________邮编: ___________)
【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 LLO/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: LLO/OVA刺激后2 h, BMDC出现短暂的TLR4 mRNA 表达上调, 其下游信号途径相关的Myd88、 Rip2、 Irak1、 Irak2、 Ikkα、 NFκB1和NFκB2 mRNA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; LLO/OVA刺激后4 h, 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κB2 mRNA均出现表达上调, IFNγ、 TNFβ和CD40 mRNA也上调表达。 LLO/OVA刺激后24 h, 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: LLO/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。
【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
[Abstract] AIM: To investigate the relationship between the activation of pattern recognition receptors and the cytokines expression of dendritic cells. METHODS: After bone marrowderived dendritic cells (BMDCs) were pulsed by LLO/OVA, the mRNA expression of pattern recognition receptors and the downstream NFκB signal pathway associated molecules were detected by microarray hybridization and RTPCR; the expression of costimulatory molecules, MHC class Ⅱ and cytokines were determined by flow cytometry and ELISA. RESULTS: After BMDCs were pulsed by LLO/OVA for 2 h, the levels of TLR4, Myd88, Rip2, Irak1, Irak2, Ikkα, NFκB1 and NFκB2 mRNA upregulated; and the levels of Icam1, IL1a, IL1b, IL6 and TNFα mRNA upregulated also; after BMDCs were pulsed by LLO/OVA for 4 h, the levels of Card4(Nod1), Rip2, Ikkβ, 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 supernatant of BMDCs pulsed by LLO/OVA at 24 h. CONCLUSION: binant LLO/OVA induces maturation and expression of c