title: Oxidized phagosomal NOX2 complex is replenished from lysosomes.
abstract: In dendritic cells, the NADPH oxidase 2 complex (NOX2) is recruited to the phagosomal membrane during antigen uptake. NOX2 produces reactive oxygen species (ROS) in the lumen of the phagosome that kill ingested pathogens, delay antigen breakdown and alter the peptide repertoire for presentation to T cells. How the integral membrane component of NOX2, cytochromeb558(which comprises CYBB and CYBA), traffics to phagosomes is incompletely understood. In this study, we show in dendritic cells derived from human blood-isolated monocytes that cytochromeb558is initially recruited to the phagosome from the plasma membrane during phagosome formation. Cytochromeb558also traffics from a lysosomal pool to phagosomes and this is required to replenish oxidatively damaged NOX2. We identified syntaxin-7, SNAP23 and VAMP8 as the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins mediating this process. Our data describe a key mechanism of how dendritic cells sustain ROS production after antigen uptake that is required to initiate T cell responses.
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen 6525 GA, The Netherlands.
authors.forename: Ilse
authors.identifier: 0000-0002-3589-5006
authors.initials: I
authors.lastname: Dingjan
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen 6525 GA, The Netherlands.
authors.forename: Peter T A
authors.identifier: 0000-0003-2228-2822
authors.initials: PT
authors.lastname: Linders
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen 6525 GA, The Netherlands.
authors.forename: Luuk
authors.initials: L
authors.lastname: van den Bekerom
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen 6525 GA, The Netherlands.
authors.forename: Maksim V
authors.initials: MV
authors.lastname: Baranov
authors.affiliation: Department of Neurobiology, Max-Planck Institute for Biophysical Chemistry, Göttingen 37077, Germany.
authors.forename: Partho
authors.initials: P
authors.lastname: Halder
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen 6525 GA, The Netherlands.
authors.forename: Martin
authors.initials: M
authors.lastname: Ter Beest
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen 6525 GA, The Netherlands Geert.vandenBogaart@Radboudumc.nl.
authors.forename: Geert
authors.initials: G
authors.lastname: van den Bogaart
mesh_terms: D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D003573:Cytochrome b Group / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D052118:Lysosomal-Associated Membrane Protein 1 / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008954:Models, Biological; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism*; D010084:Oxidation-Reduction; D010588:Phagosomes / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins / Q000378:metabolism; D050767:Qc-SNARE Proteins / Q000378:metabolism; D050683:R-SNARE Proteins / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / Q000378:metabolism
chemical_list: D003573:Cytochrome b Group; D052118:Lysosomal-Associated Membrane Protein 1; D008562:Membrane Glycoproteins; D010716:Phosphatidylinositols; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D050683:R-SNARE Proteins; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; C100493:SNAP23 protein, human; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; C112938:VAMP8 protein, human; C036888:cytochrome b558; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases
keywords: Dendritic cell; NOX2; Phagocytosis; Phagosome maturation; Reactive oxygen species; SNARE