title: Chimeric Vaccine Stimulation of Human Dendritic Cell Indoleamine 2, 3-Dioxygenase Occurs via the Non-Canonical NF-κB Pathway.
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.
authors.affiliation: Center for Health Disparities and Molecular Medicine, Department of Basic Sciences, Loma Linda University School of Medicine, Loma Linda, California, United States of America.|Department of Molecular Biology, Chonbuk National University, Jeon-Ju, Republic of Korea.
authors.forename: Nan-Sun
authors.initials: NS
authors.lastname: Kim
authors.affiliation: Center for Health Disparities and Molecular Medicine, Department of Basic Sciences, Loma Linda University School of Medicine, Loma Linda, California, United States of America.|Loma Linda University School of Medicine, Department of Basic Sciences, Division of Physiology, Loma Linda, California, United States of America.
authors.forename: Jacques C
authors.initials: JC
authors.lastname: Mbongue
authors.affiliation: Center for Health Disparities and Molecular Medicine, Department of Basic Sciences, Loma Linda University School of Medicine, Loma Linda, California, United States of America.|Loma Linda University School of Medicine, Department of Basic Sciences, Division of Biochemistry, Loma Linda, California, United States of America.
authors.forename: Dequina A
authors.initials: DA
authors.lastname: Nicholas
authors.affiliation: Center for Health Disparities and Molecular Medicine, Department of Basic Sciences, Loma Linda University School of Medicine, Loma Linda, California, United States of America.|Loma Linda University School of Medicine, Department of Earth and Biological Sciences, Loma Linda, California, United States of America.
authors.forename: Grace E
authors.initials: GE
authors.lastname: Esebanmen
authors.affiliation: Loma Linda University School of Medicine, Department of Basic Sciences, Division of Biochemistry, Loma Linda, California, United States of America.
authors.forename: Juli J
authors.initials: JJ
authors.lastname: Unternaehrer
authors.affiliation: Endocrinology Section, JL Pettis Memorial VA Medical Center, Loma Linda, California, United States of America.
authors.forename: Anthony F
authors.initials: AF
authors.lastname: Firek
authors.affiliation: Center for Health Disparities and Molecular Medicine, Department of Basic Sciences, Loma Linda University School of Medicine, Loma Linda, California, United States of America.|Loma Linda University School of Medicine, Department of Basic Sciences, Division of Biochemistry, Loma Linda, California, United States of America.
authors.forename: William H R
authors.initials: WH
authors.lastname: Langridge
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015551:Autoimmunity / Q000187:drug effects; D001483:Base Sequence; D002772:Cholera Toxin / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D005786:Gene Expression Regulation; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011384:Proinsulin / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction; D047992:TNF Receptor-Associated Factor 2 / Q000494:pharmacology; D048008:TNF Receptor-Associated Factor 3 / Q000494:pharmacology; D048029:TNF Receptor-Associated Factor 6 / Q000494:pharmacology; D014612:Vaccines / Q000008:administration & dosage*; D000097782:NF-kappaB-Inducing Kinase
chemical_list: D002772:Cholera Toxin; D051550:I-kappa B Kinase; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D016328:NF-kappa B; D011384:Proinsulin; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D047992:TNF Receptor-Associated Factor 2; D048008:TNF Receptor-Associated Factor 3; D048029:TNF Receptor-Associated Factor 6; D014612:Vaccines; D000097782:NF-kappaB-Inducing Kinase; C576710:IDO1 protein, human; C090656:TRAF3 protein, human