Expression of IL-12-related molecules in human intestinal microvascular endothelial cells is regulated by TLR3
Expression of IL-12-related molecules in human intestinal microvascular endothelial cells is regulated by TLR3
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DOI：
10.1152/ajpgi.00142.2007
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发表时间：
2007-12-01
影响因子：
4.5
通讯作者：
Maaser, Christian
中科院分区：
文献类型：
--
作者：
Heidemann, Jan;Ruether, Christoph;Maaser, Christian
IL-12家族的成员构成IL-12、-23和-27的亚单位。这些ILS在调节细胞介导的免疫反应和人类炎症性肠病的动物模型中是关键的介质。最近的研究表明，肠道内皮细胞可能是细菌感知入侵病原体的第二道防线。本研究的目的是检测肠内皮细胞(HIMEC)产生IL-12家族成员的情况。用促炎剂(肿瘤坏死因子-α、干扰素-γ、白介素1-β)和微生物抗原[脂多糖、脂磷壁酸、肽聚糖、CpG-DNA、鞭毛蛋白、聚(I：C)]刺激HIMEC。用实时荧光定量RT-PCR、免疫组化、流式细胞仪和免疫印迹分析检测IL-12家族成员和TLR3在HIMEC中的表达。HIMEC表达EB病毒诱导基因3(EBI3)、IL-12p35和IL-23p19，未检测到IL-12p40和IL-27p28的表达。已知的利用核因子-kappa B途径的促炎因子诱导的诱导作用最强，而核因子-kappaB抑制剂可抑制EBI3和IL-23p19的表达。HIMEC显示TLR3的调控表达。黏附和移行实验显示HIMEC刺激后出现促炎反应。HIMEC能够产生IL-12家族成员作为对微生物刺激的反应。TLR3激动剂Poly(I：C)在体外可增强HIMEC中白细胞的黏附。我们的数据表明，肠微血管系统对肠道内皮细胞表达的TLR3配体有反应，从而增加了适应性免疫和白细胞募集的调节。
Members of the interleukin (IL)-12 family constitute subunits of IL-12, -23, and -27. These ILs represent pivotal mediators in the regulation of cell-mediated immune responses and in animal models of human inflammatory bowel disease. Recent work has suggested that intestinal endothelial cells might serve as a second line of defense in bacterial sensing of invading pathogens. The purpose of this study was to examine the production of IL-12 family members in intestinal endothelial cells (HIMEC). HIMEC were stimulated with proinflammatory agents (TNF-alpha, IFN-gamma, IL-1 beta) and microbial antigens [LPS, lipoteichoic acid, peptidoglycan, CpG-DNA, flagellin, poly(I:C)]. Expression of IL-12 family members and of Toll-like receptor (TLR)3 in HIMEC was assessed by real-time RT-PCR, immunostaining, flow cytometry, and immunoblot analysis. HIMEC display an induction of Epstein-Barr virusinduced gene 3 (EBI3), IL-12p35, and IL-23p19, whereas no expression of IL-12p40 and IL-27p28 was detectable. The strongest induction was induced by proinflammatory factors known to utilize the NF-kappa B pathway, and expression of EBI3 and IL-23p19 was diminished by an NF-kappa B inhibitor. HIMEC display regulated expression of TLR3. Adhesion and transmigration assays showed proinflammatory responses after HIMEC stimulation. HIMEC are capable of producing IL-12 family members as a response to microbial stimuli. The TLR3 agonist, poly( I: C), was shown to enhance leukocyte adhesion in vitro in HIMEC. Our data suggest that the intestinal microvasculature is responsive to ligands of TLR3 expressed on intestinal endothelial cells, thereby adding to the regulation of adaptive immunity and leukocyte recruitment.