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Rab27a regulates phagosomal pH and NADPH oxidase recruitment to dendritic cell phagosomes

Nature Cell Biology volume 9, pages 367–378 (2007)Cite this article

Abstract

To prevent excessive degradation of internalized antigens, which could destroy the peptides recognized by T lymphocytes, dendritic cells have developed several strategies that limit proteolytic activity in phagosomes. The recruitment of the NADPH oxidase NOX2 prevents acidification of phagosomes, limiting antigen degradation. Here, we show that dendritic cells derived from Rab27a-deficient ashen mice show increased phagosome acidification and antigen degradation, causing a defect in antigen cross-presentation. Enhanced acidification results from a delay in the recruitment to phagosomes of a subset of lysosome-related organelles containing the membrane subunits of NOX2. The Rab27a-dependent recruitment of these “inhibitory lysosome-related organelles” to phagosomes continuously limits acidification and degradation of ingested particles in dendritic cells, thus promoting antigen cross-presentation.

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Figure 1: Rab27a is specifically expressed in wild-type (WT) dendritic cells (DCs) and is efficiently recruited to dendritic-cell phagosomes.
Figure 2: Decreased cross-presentation in Rab27a-deficient dendritic cells.
Figure 3: Excessive phagosomal acidification in Rab27a-deficient dendritic cells causes inhibition in cross-presentation.
Figure 4: Increased antigen degradation in Rab27a-deficient dendritic cells causes inefficient cross-presentation.
Figure 5: NOX2 colocalizes with Rab27a.
Figure 6: NOX2 is present in lysosome-related organelles.
Figure 7: Recruitment of lysosome-related organelles to phagosomes is delayed in the absence of Rab27a.
Figure 8: NOX2 is inefficiently recruited and activated in Rab27a-deficient dendritic cells.

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Acknowledgements

We thank R. Anders and D. Barral for providing bone marrow cells during the early stages of this work and H. Graham and M. Mules for mouse breeding. We are grateful to F. Sepulveda and B. Manoury for helpful discussion and for their help with the in vitro degradation assays and C. Watts for providing the tetanus toxin C-fragment. We thank J.-B. Sibarita for assistence with the image processing and S. Dogniaux, P. Benaroch, P. Guermonprez and all the members of U653 INSERM for help with reagents and discussions. We also acknowledge J. Calafat and H. Janssen for the gift of 7D5 antibody (anti-cytb558), and I. Cruz Moura and H. Schild for HY long peptide design and validation. We are grateful to the Ligue National de Lutte Contre le Cancer (LLNC), the Welcome Trust and BBSRC and Fondation Bettencourt, for support. A.S. was supported by LLNC, INSERM and Marie Curie Fellowship Program, and C.J. was supported by the Institut Curie, INSERM and Fondation pour la Recherche Medicale.

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  1. Carolina Jancic and Ariel Savina: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut Curie, INSERM U653, Immunité et Cancer, 26 rue d'Ulm, 75248, Paris, Cedex 05, France

    Carolina Jancic, Ariel Savina & Sebastian Amigorena

  2. Molecular and Cellular Medicine, NHLI, Imperial College London, Exhibition Rd, London, SW7 2AZ, UK

    Christina Wasmeier, Tanya Tolmachova & Miguel C. Seabra

  3. INSERM U773,Centre de Recherche Biomedicale Bichat Beaujon CRB3,

    Jamel El-Benna, Pham My-Chan Dang & Maire-Anne Gougerot-Pocidalo

  4. Université Paris 7 Denis Diderot, site Bichat, UMRS 773, Paris, F-75018, France

    Jamel El-Benna, Pham My-Chan Dang & Maire-Anne Gougerot-Pocidalo

  5. Department of Immunology University of Tuebingen, Auf der Morgenstelle 15, Tuebingen, 72076, Germany

    Steve Pascolo

  6. Institut Curie, CNRS-UMR144, Structure et Compartiments Membranaires, 26 rue d'Ulm, 75248, Paris, Cedex 05, France

    Graça Raposo

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  1. Carolina Jancic
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Correspondence to Sebastian Amigorena.

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Jancic, C., Savina, A., Wasmeier, C. et al. Rab27a regulates phagosomal pH and NADPH oxidase recruitment to dendritic cell phagosomes. Nat Cell Biol 9, 367–378 (2007). https://doi.org/10.1038/ncb1552

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