PMID- 30808727 OWN - NLM STAT- MEDLINE DCOM- 20200116 LR - 20210325 IS - 1535-9484 (Electronic) IS - 1535-9476 (Print) IS - 1535-9476 (Linking) VI - 18 IP - 5 DP - 2019 May TI - Spatiotemporal Changes of the Phagosomal Proteome in Dendritic Cells in Response to LPS Stimulation. PG - 909-922 LID - S1535-9476(20)31602-9 [pii] LID - 10.1074/mcp.RA119.001316 [doi] AB - Dendritic cells (DCs) are professional phagocytes that use innate sensing and phagocytosis to internalize and degrade self as well as foreign material, such as pathogenic bacteria, within phagosomes. These intracellular compartments are equipped to generate antigenic peptides that serve as source for antigen presentation to T cells initiating adaptive immune responses. The phagosomal proteome of DCs is only partially studied and is highly dynamic as it changes during phagosome maturation, when phagosomes sequentially interact with endosomes and lysosomes. In addition, the activation status of the phagocyte can modulate the phagosomal composition and is able to shape phagosomal functions.In this study, we determined spatiotemporal changes of the proteome of DC phagosomes during their maturation and compared resting and lipopolysaccharide (LPS)-stimulated bone marrow-derived DCs by label-free, quantitative mass spectrometry. Ovalbumin-coupled latex beads were used as phagocytosis model system and revealed that LPS-treated DCs show decreased recruitment of proteins involved in phagosome maturation, such as subunits of the vacuolar proton ATPase, cathepsin B, D, S, and RAB7. In contrast, those phagosomes were characterized by an increased recruitment of proteins involved in antigen cross-presentation, e.g. different subunits of MHC I molecules, the proteasome and tapasin, confirming the observed increase in cross-presentation efficacy in those cells. Further, several proteins were identified that were not previously associated with phagosomal functions. Hierarchical clustering of phagosomal proteins demonstrated that their acquisition to DC phagosomes is not only dependent on the duration of phagosome maturation but also on the activation state of DCs. Thus, our study provides a comprehensive overview of how DCs alter their phagosome composition in response to LPS, which has profound impact on the initiation of efficient immune responses. CI - (c) 2019 Pauwels et al. FAU - Pauwels, Anne-Marie AU - Pauwels AM AD - From the double daggerUnit of Molecular Signal Transduction in Inflammation, VIB Center for Inflammation Research, Ghent, Belgium;; section signDepartment of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. FAU - Hartlova, Anetta AU - Hartlova A AD - paragraph signInstitute for Cell and Molecular Biosciences, Newcastle University, Newcastle-upon-Tyne, UK. FAU - Peltier, Julien AU - Peltier J AD - paragraph signInstitute for Cell and Molecular Biosciences, Newcastle University, Newcastle-upon-Tyne, UK. FAU - Driege, Yasmine AU - Driege Y AD - From the double daggerUnit of Molecular Signal Transduction in Inflammation, VIB Center for Inflammation Research, Ghent, Belgium;; section signDepartment of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. FAU - Baudelet, Griet AU - Baudelet G AD - From the double daggerUnit of Molecular Signal Transduction in Inflammation, VIB Center for Inflammation Research, Ghent, Belgium;; section signDepartment of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. FAU - Brodin, Priscille AU - Brodin P AD - ||Univ. Lille, CNRS, INSERM, CHU Lille, Institut Pasteur de Lille, U1019-UMR 8204-CIIL-Center for Infection and Immunity of Lille, Lille, France. FAU - Trost, Matthias AU - Trost M AD - paragraph signInstitute for Cell and Molecular Biosciences, Newcastle University, Newcastle-upon-Tyne, UK. FAU - Beyaert, Rudi AU - Beyaert R AD - From the double daggerUnit of Molecular Signal Transduction in Inflammation, VIB Center for Inflammation Research, Ghent, Belgium;; section signDepartment of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. FAU - Hoffmann, Eik AU - Hoffmann E AD - From the double daggerUnit of Molecular Signal Transduction in Inflammation, VIB Center for Inflammation Research, Ghent, Belgium;; section signDepartment of Biomedical Molecular Biology, Ghent University, Ghent, Belgium;; ||Univ. Lille, CNRS, INSERM, CHU Lille, Institut Pasteur de Lille, U1019-UMR 8204-CIIL-Center for Infection and Immunity of Lille, Lille, France. LA - eng GR - MC_UP_A500_1020/MRC_/Medical Research Council/United Kingdom GR - MC_UU_12016/5/MRC_/Medical Research Council/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190226 PL - United States TA - Mol Cell Proteomics JT - Molecular & cellular proteomics : MCP JID - 101125647 RN - 0 (Lipopolysaccharides) RN - 0 (Proteome) SB - IM MH - Animals MH - Dendritic Cells/drug effects/*metabolism MH - Kinetics MH - Lipopolysaccharides/*pharmacology MH - Mice, Inbred C57BL MH - Phagosomes/drug effects/*metabolism MH - Proteome/*metabolism MH - Proteomics MH - Time Factors PMC - PMC6495253 OTO - NOTNLM OT - Cell biology* OT - Cellular organelles* OT - Endocytosis* OT - Immunology* OT - Inflammatory response OT - Label-free quantification OT - Protein Identification* OT - dendritic cell OT - phagocytosis OT - phagosome maturation EDAT- 2019/02/28 06:00 MHDA- 2020/01/17 06:00 PMCR- 2020/05/01 CRDT- 2019/02/28 06:00 PHST- 2019/01/10 00:00 [received] PHST- 2019/02/23 00:00 [revised] PHST- 2019/02/28 06:00 [pubmed] PHST- 2020/01/17 06:00 [medline] PHST- 2019/02/28 06:00 [entrez] PHST- 2020/05/01 00:00 [pmc-release] AID - S1535-9476(20)31602-9 [pii] AID - RA119.001316 [pii] AID - 10.1074/mcp.RA119.001316 [doi] PST - ppublish SO - Mol Cell Proteomics. 2019 May;18(5):909-922. doi: 10.1074/mcp.RA119.001316. Epub 2019 Feb 26.