1. Chimeric Vaccine Stimulation of Human Dendritic Cell Indoleamine 2, 3-Dioxygenase Occurs via the Non-Canonical NF-κB Pathway.
- Author
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Kim NS, Mbongue JC, Nicholas DA, Esebanmen GE, Unternaehrer JJ, Firek AF, and Langridge WH
- Subjects
- Amino Acid Sequence, Animals, Autoimmunity drug effects, Base Sequence, Cholera Toxin biosynthesis, Cholera Toxin genetics, Cholera Toxin immunology, Dendritic Cells immunology, Dendritic Cells pathology, Diabetes Mellitus, Type 1 genetics, Diabetes Mellitus, Type 1 immunology, Diabetes Mellitus, Type 1 pathology, Gene Expression Regulation, Humans, I-kappa B Kinase genetics, I-kappa B Kinase immunology, I-kappa B Kinase metabolism, Indoleamine-Pyrrole 2,3,-Dioxygenase genetics, Indoleamine-Pyrrole 2,3,-Dioxygenase metabolism, Mice, Mice, Inbred NOD, Molecular Sequence Data, NF-kappa B genetics, NF-kappa B metabolism, Proinsulin biosynthesis, Proinsulin genetics, Proinsulin immunology, Protein Serine-Threonine Kinases genetics, Protein Serine-Threonine Kinases immunology, Protein Serine-Threonine Kinases metabolism, RNA, Small Interfering genetics, RNA, Small Interfering metabolism, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins immunology, Signal Transduction, TNF Receptor-Associated Factor 2 pharmacology, TNF Receptor-Associated Factor 3 pharmacology, TNF Receptor-Associated Factor 6 pharmacology, NF-kappaB-Inducing Kinase, Dendritic Cells drug effects, Diabetes Mellitus, Type 1 therapy, Indoleamine-Pyrrole 2,3,-Dioxygenase immunology, NF-kappa B immunology, Vaccines administration & dosage
- Abstract
A chimeric protein vaccine composed of the cholera toxin B subunit fused to proinsulin (CTB-INS) was shown to suppress type 1 diabetes onset in NOD mice and upregulate biosynthesis of the tryptophan catabolic enzyme indoleamine 2, 3-dioxygenase (IDO1) in human dendritic cells (DCs). Here we demonstrate siRNA inhibition of the NF-κB-inducing kinase (NIK) suppresses vaccine-induced IDO1 biosynthesis as well as IKKα phosphorylation. Chromatin immunoprecipitation (ChIP) analysis of CTB-INS inoculated DCs showed that RelB bound to NF-κB consensus sequences in the IDO1 promoter, suggesting vaccine stimulation of the non-canonical NF-κB pathway activates IDO1 expression in vivo. The addition of Tumor Necrosis Factor Associated Factors (TRAF) TRAF 2, 3 and TRAF6 blocking peptides to vaccine inoculated DCs was shown to inhibit IDO1 biosynthesis. This experimental outcome suggests vaccine activation of the TNFR super-family receptor pathway leads to upregulation of IDO1 biosynthesis in CTB-INS inoculated dendritic cells. Together, our experimental data suggest the CTB-INS vaccine uses a TNFR-dependent signaling pathway of the non-canonical NF-κB signaling pathway resulting in suppression of dendritic cell mediated type 1 diabetes autoimmunity.
- Published
- 2016
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