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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Autoimmunity / drug effects
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Base Sequence
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Cholera Toxin / biosynthesis
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Cholera Toxin / genetics
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Cholera Toxin / immunology
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Dendritic Cells / drug effects*
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Dendritic Cells / immunology
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Dendritic Cells / pathology
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Diabetes Mellitus, Type 1 / genetics
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Diabetes Mellitus, Type 1 / immunology
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Diabetes Mellitus, Type 1 / pathology
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Diabetes Mellitus, Type 1 / therapy*
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Gene Expression Regulation
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Humans
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I-kappa B Kinase / genetics
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I-kappa B Kinase / immunology
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I-kappa B Kinase / metabolism
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Indoleamine-Pyrrole 2,3,-Dioxygenase / genetics
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Indoleamine-Pyrrole 2,3,-Dioxygenase / immunology*
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Indoleamine-Pyrrole 2,3,-Dioxygenase / metabolism
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Mice
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Mice, Inbred NOD
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Molecular Sequence Data
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NF-kappa B / genetics
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NF-kappa B / immunology*
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NF-kappa B / metabolism
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NF-kappaB-Inducing Kinase
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Proinsulin / biosynthesis
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Proinsulin / genetics
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Proinsulin / immunology
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / immunology
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Protein Serine-Threonine Kinases / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Signal Transduction
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TNF Receptor-Associated Factor 2 / pharmacology
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TNF Receptor-Associated Factor 3 / pharmacology
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TNF Receptor-Associated Factor 6 / pharmacology
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Vaccines / administration & dosage*
Substances
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IDO1 protein, human
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Indoleamine-Pyrrole 2,3,-Dioxygenase
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NF-kappa B
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RNA, Small Interfering
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Recombinant Fusion Proteins
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TNF Receptor-Associated Factor 2
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TNF Receptor-Associated Factor 3
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TNF Receptor-Associated Factor 6
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TRAF3 protein, human
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Vaccines
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Cholera Toxin
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Proinsulin
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Protein Serine-Threonine Kinases
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I-kappa B Kinase