Regulation of IL-8 and IL-1β expression in Crohn's disease associated NOD2/CARD15 mutations
Regulation of IL-8 and IL-1β expression in Crohn's disease associated NOD2/CARD15 mutations
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DOI：
10.1093/hmg/ddh182
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发表时间：
2004-08-15
影响因子：
3.5
通讯作者：
Cho, JH
中科院分区：
文献类型：
--
作者：
Li, J;Moran, T;Cho, JH
克罗恩病（CD）是一种影响胃肠道的慢性炎症。NOD 2/CARD 15基因内的三个突变（Arg 702 Trp、Gly 908 Arg和Leu 1007 fsinsC）增加CD易感性。在这里，我们定义了细胞因子的调节，在原代人单核细胞，胞壁酰二肽（MDP），最小的NOD 2/CARD 15激活成分的肽聚糖。通过微阵列，MDP诱导广泛的转录物，包括白细胞介素1 β（IL-1 β）和白细胞介素8（IL-8）。Leu 1007 fsinsC纯合子表现出对MDP的转录反应降低。电迁移率变动分析表明，MDP诱导的NF-κ B活化是通过p50和p65亚基介导的，而不是RelB或c-Rel。在野生型个体中，与低剂量（10 ng/ml）MDP相比，MDP诱导的IL-8蛋白表达对高剂量（1 μ g/ml）的反应更大。在低MDP剂量下，在所有纯合子中，我们没有观察到IL-8蛋白的诱导。在高剂量MDP下，Leu 1007 fsinsC纯合子未显示诱导。在Gly 908 Arg和Arg 702 Trp纯合子中观察到IL-8蛋白的适度诱导，表明CD相关突变的不同MDP敏感性。在野生型健康对照、CD和溃疡性结肠炎个体中，单独使用低剂量MDP和TNF α仅导致适度的IL-1 β蛋白诱导。MDP与TNF α联合使用，可协同诱导IL-1 β分泌。在Leu 1007 fsinsC纯合子中，尽管IL-1 β mRNA有明显的诱导作用，但IL-1 β分泌存在严重缺陷。这些发现证明了MDP和TNF α处理对IL-1 β分泌的NOD 2/CARD 15途径的转录后依赖性。总之，这些研究表明，先天免疫MDP的信号缺陷可能是一些CD患者发病机制中的一个重要的潜在缺陷。
Crohn's disease (CD) is a chronic inflammation affecting the gastrointestinal tract. Three mutations (Arg702Trp, Gly908Arg and Leu1007fsinsC) within the NOD2/CARD15 gene increase CD susceptibility. Here, we define cytokine regulation in primary human mononuclear cells, with muramyl dipeptide (MDP), the minimal NOD2/CARD15 activating component of peptidoglycan. By microarray, MDP induces a broad array of transcripts, including interleukin 1beta (IL-1beta) and interleukin 8 (IL-8). Leu1007fsinsC homozygotes demonstrated decreased transcriptional response to MDP. Electromobility shift assay demonstrated that MDP-induced NF-kappaB activation is mediated via p50 and p65 subunits, but not RelB or c-Rel. In wild-type individuals, MDP-induced IL-8 protein expression with a greater response to high dose (1 mug/ml) compared with low-dose (10 ng/ml) MDP. At low MDP doses, in all homozygotes, we observed no induction of IL-8 protein. With high doses of MDP, Leu1007fsinsC homozygotes showed no induction. Modest induction of IL-8 protein was observed in Gly908Arg and Arg702Trp homozygotes, indicating varying MDP sensitivity of the CD-associated mutations. In wild-type healthy control, CD and ulcerative colitis individuals, low-dose MDP and TNFalpha alone results in only modest IL-1beta protein induction. With MDP plus TNFalpha, there is a synergistic induction of IL-1beta secretion. In Leu1007fsinsC homozygotes, there is a profound defect in IL-1beta secretion, despite marked induction of IL-1beta mRNA. These findings demonstrate post-transcriptional dependency on the NOD2/CARD15 pathway for IL-1beta secretion with MDP and TNFalpha treatment. Taken together, these studies suggest that a signaling defect of innate immunity to MDP may be an essential underlying defect in the pathogenesis of some CD patients.