Journal Article

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

Hulin, Philippe, Nedellec, Steven, Ménager, Jérémie, Ebstein, Frédéric, Duverger, Eric, Kloetzel, Peter-Michael, Jotereau, Francine, Lehmann, Andrea, Oger, Romain, Guilloux, Yannick

2014

Overview

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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adenosine Triphosphatases / Adenosine Triphosphatases - metabolism / Amino acids / Antigen Presentation / Antigen Presentation - immunology / Antigen-presenting cells / Antigens / Antigens - chemistry / Antigens - immunology / Antigens, Neoplasm / Antigens, Neoplasm - chemistry / Antigens, Neoplasm - immunology / ATP-Binding Cassette Transporters / ATP-Binding Cassette Transporters - metabolism / Biochemistry / Biology / Cancer / Cancer vaccines / CD8 antigen / Cell Cycle Proteins / Cell Cycle Proteins - metabolism / Confocal microscopy / Cross-Priming / Cross-Priming - immunology / Cytoplasm / Cytosol / Cytotoxicity / Dendritic Cells / Dendritic Cells - immunology / Dendritic Cells - metabolism / Endocytosis / Endocytosis - immunology / Endoplasmic reticulum / Endoplasmic Reticulum-Associated Degradation / Endosomes / Endosomes - metabolism / Health aspects / Histocompatibility antigen HLA / HLA antigens / Human health and pathology / Humans / Immune system / Immunology / Immunotherapy / Kinetics / Life Sciences / Lymphocytes / Lymphocytes T / Lysosomes / Lysosomes - metabolism / Machinery and equipment / Major histocompatibility complex / MART-1 Antigen / MART-1 Antigen - chemistry / MART-1 Antigen - immunology / Membrane Proteins / Membrane Proteins - metabolism / Microscopy / Models, Biological / Peptides / Peptides - chemical synthesis / Peptides - chemistry / Peptides - immunology / Proteasome Endopeptidase Complex / Proteasome Endopeptidase Complex - metabolism / Proteasomes / Protein Transport / Proteins / Quality control / SEC Translocation Channels / Subunit structure / T cell receptors / T cells / Translocation / Tumors / Vaccination / Vaccines / Valosin Containing Protein