title: Cross-presentation of synthetic long peptides by human dendritic cells: a process dependent on ERAD component p97/VCP but Not sec61 and/or Derlin-1.
abstract: Antitumor vaccination using synthetic long peptides (SLP) is an additional therapeutic strategy currently under development. It aims to activate tumor-specific CD8(+) CTL by professional APCs such as DCs. DCs can activate T lymphocytes by MHC class I presentation of exogenous antigens - a process referred to as "cross-presentation". Until recently, the intracellular mechanisms involved in cross-presentation of soluble antigens have been unclear. Here, we characterize the cross-presentation pathway of SLP Melan-A16-40 containing the HLA-A2-restricted epitope26-35 (A27L) in human DCs. Using confocal microscopy and specific inhibitors, we show that SLP16-40 is rapidly taken up by DC and follows a classical TAP- and proteasome-dependent cross-presentation pathway. Our data support a role for the ER-associated degradation machinery (ERAD)-related protein p97/VCP in the transport of SLP16-40 from early endosomes to the cytoplasm but formally exclude both sec61 and Derlin-1 as possible retro-translocation channels for cross-presentation. In addition, we show that generation of the Melan-A26-35 peptide from the SLP16-40 was absolutely not influenced by the proteasome subunit composition in DC. Altogether, our findings propose a model for cross-presentation of SLP which tends to enlarge the repertoire of potential candidates for retro-translocation of exogenous antigens to the cytosol.
authors.affiliation: INSERM U892, Nantes, France ; Université de Nantes, Nantes, France ; CNRS, UMR 6299, Nantes, France.
authors.forename: Jérémie
authors.initials: J
authors.lastname: Ménager
authors.affiliation: Institut of Biochemistry, Charité University Hospital, Humboldt University, Berlin, Germany.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Ebstein
authors.affiliation: INSERM U892, Nantes, France ; Université de Nantes, Nantes, France ; CNRS, UMR 6299, Nantes, France.
authors.forename: Romain
authors.initials: R
authors.lastname: Oger
authors.affiliation: INSERM U892, Nantes, France ; CNRS, UMR 6299, Nantes, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Hulin
authors.affiliation: INSERM U892, Nantes, France ; CNRS, UMR 6299, Nantes, France.
authors.forename: Steven
authors.initials: S
authors.lastname: Nedellec
authors.affiliation: Glycobiochimie, ICOA, Université d'Orléans, Orléans, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Duverger
authors.affiliation: Institut of Biochemistry, Charité University Hospital, Humboldt University, Berlin, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Lehmann
authors.affiliation: Institut of Biochemistry, Charité University Hospital, Humboldt University, Berlin, Germany.
authors.forename: Peter-Michael
authors.initials: PM
authors.lastname: Kloetzel
authors.affiliation: INSERM U892, Nantes, France ; Université de Nantes, Nantes, France ; CNRS, UMR 6299, Nantes, France.
authors.forename: Francine
authors.initials: F
authors.lastname: Jotereau
authors.affiliation: INSERM U892, Nantes, France ; Université de Nantes, Nantes, France ; CNRS, UMR 6299, Nantes, France.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Guilloux
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D017951:Antigen Presentation / Q000276:immunology*; D000941:Antigens / Q000737:chemistry / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000276:immunology; D018797:Cell Cycle Proteins / Q000378:metabolism; D045142:Cross-Priming / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D004705:Endocytosis / Q000276:immunology; D060746:Endoplasmic Reticulum-Associated Degradation; D011992:Endosomes / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008247:Lysosomes / Q000378:metabolism; D058965:MART-1 Antigen / Q000737:chemistry / Q000276:immunology; D008565:Membrane Proteins / Q000378:metabolism; D008954:Models, Biological; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D021381:Protein Transport; D000069816:SEC Translocation Channels; D000074405:Valosin Containing Protein
chemical_list: D018528:ATP-Binding Cassette Transporters; D000941:Antigens; D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; C487709:DERL1 protein, human; D058965:MART-1 Antigen; D008565:Membrane Proteins; D010455:Peptides; D000069816:SEC Translocation Channels; C477327:transporter associated with antigen processing (TAP); D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein