Anti-Dengue Virus Nonstructural Protein 1 Antibodies Cause NO-Mediated Endothelial Cell Apoptosis via Ceramide-Regulated Glycogen Synthase Kinase-3β and NF-κB Activation
Anti-Dengue Virus Nonstructural Protein 1 Antibodies Cause NO-Mediated Endothelial Cell Apoptosis via Ceramide-Regulated Glycogen Synthase Kinase-3β and NF-κB Activation
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DOI：
10.4049/jimmunol.1201976
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发表时间：
2013-08-15
影响因子：
4.4
通讯作者：
Lin, Yee-Shin
中科院分区：
文献类型：
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作者：
Chen, Chia-Ling;Lin, Chiou-Feng;Lin, Yee-Shin
登革病毒（DENV）感染的免疫病理机制涉及血小板减少症、凝血病和血管病引起的出血综合征。我们提出了一种分子模拟机制，其中针对DENV非结构蛋白1（NS 1）的Ab与人内皮细胞交叉反应，并引起NF-κ B调节的免疫激活和NO介导的凋亡。然而，在抗DENV NS 1抗体与内皮细胞结合后导致NF-κ B活化的信号传导途径尚未解决。在这项研究中，我们发现，抗DENV NS 1抗体引起脂质筏样结构的形成，并且通过甲基-β-环糊精破坏脂质筏形成减少NO产生和细胞凋亡。用抗DENV NS 1 Ab处理提高了脂筏中的神经酰胺生成。酸性鞘磷脂酶（aSMase）的药理学抑制降低了抗DENV NS 1 Ab介导的神经酰胺和NO产生以及细胞凋亡。外源性神经酰胺处理诱导的诱导型NO合酶（iNOS）/NO和细胞凋亡的生物合成通过NF-κ B调节的方式。此外，神经酰胺诱导的NF-κ B活化和iNOS表达需要糖原合成酶激酶-3 β（GSK-3 β）的活化。值得注意的是，抗DENV NS 1 Ab引起GSK-3 β介导的NF-κ B活化和iNOS表达，其由aSM酶调节。此外，GSK-3 β的药理学抑制减少了被动施用抗DENV NS 1 Ab的小鼠中的肝内皮细胞凋亡。这些结果表明，抗DENV NS 1 Ab结合至内皮细胞膜，并通过涉及aSMase/神经酰胺/GSK-3 β/NF-κ B/iNOS/NO信号传导途径的机制引起NO产生和凋亡。
Immunopathogenetic mechanisms of dengue virus (DENV) infection are involved in hemorrhagic syndrome resulting from thrombocytopenia, coagulopathy, and vasculopathy. We have proposed a mechanism of molecular mimicry in which Abs against DENV nonstructural protein 1 (NS1) cross-react with human endothelial cells and cause NF-kappa B-regulated immune activation and NO-mediated apoptosis. However, the signaling pathway leading to NF-kappa B activation after the binding of anti-DENV NS1 Abs to endothelial cells is unresolved. In this study, we found that anti-DENV NS1 Abs caused the formation of lipid raftlike structures, and that disrupting lipid raft formation by methyl-beta-cyclodextrin decreased NO production and apoptosis. Treatment with anti-DENV NS1 Abs elevated ceramide generation in lipid rafts. Pharmacological inhibition of acid sphingomyelinase (aSMase) decreased anti-DENV NS1 Ab-mediated ceramide and NO production, as well as apoptosis. Exogenous ceramide treatment induced biogenesis of inducible NO synthase (iNOS)/NO and apoptosis through an NF-kappa B-regulated manner. Furthermore, activation of glycogen synthase kinase-3 beta (GSK-3 beta) was required for ceramide-induced NF-kappa B activation and iNOS expression. Notably, anti-DENV NS1 Abs caused GSK-3 beta-mediated NF-kappa B activation and iNOS expression, which were regulated by aSMase. Moreover, pharmacological inhibition of GSK-3 beta reduced hepatic endothelial cell apoptosis in mice passively administered anti-DENV NS1 Abs. These results suggest that anti-DENV NS1 Abs bind to the endothelial cell membrane and cause NO production and apoptosis via a mechanism involving the aSMase/ceramide/GSK-3 beta/NF-kappa B/iNOS/NO signaling pathway.