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Blood, 1 December 2008, Vol. 112, No. 12, pp. 4712-4722.
Prepublished online as a Blood First Edition Paper on August 5, 2008; DOI 10.1182/blood-2008-01-134791.
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PHAGOCYTES
NADPH oxidase controls phagosomal pH and antigen cross-presentation in human dendritic cells
Adriana R. Mantegazza1,
Ariel Savina1,
Mónica Vermeulen2,
Laura Pérez3,
Jorge Geffner2,
Olivier Hermine4,
Sergio D. Rosenzweig3,
Florence Faure1, and
Sebastián Amigorena1
1 Inserm U653 Immunité et cancer, Institut Curie, Paris, France;
2 Instituto de Investigaciones Hematológicas, Academia Nacional de Medicina, Buenos Aires, Argentina;
3 Servicio de Immunología, Hospital Garrahan, Buenos Aires, Argentina; and
4 Centre National de Recherche Scientifique (CNRS) UMR 8147, Hôpital Necker-Enfants-Malades, Paris, France
The phagocyte NADPH oxidase (NOX2) is critical for the bactericidal activity of phagocytic cells and plays a major role in innate immunity. We showed recently that NOX2 activity in mouse dendritic cells (DCs) prevents acidification of phagosomes, promoting antigen cross-presentation. Inorder to investigate the role of NOX2 in the regulation of the phagosomal pH in human DCs, we analyzed the production of reactive oxygen species (ROS) and the phagosomal/endosomal pH in monocyte-derived DCs and macrophages (MØs) from healthy donors or patients with chronic granulomatous disease (CGD). As expected, we found that human MØs acidify their phagosomes more efficiently than human DCs. Accordingly, the expression of the vacuolar proton ATPase (V-H+-ATPase) was higher in MØs than in DCs. Phagosomal ROS production, however, was also higher in MØs than in DCs, due to higher levels of gp91phox expression and recruitment to phagosomes. In contrast, in the absence of active NOX2, the phagosomal and endosomal pH decreased. Both in the presence of a NOX2 inhibitor and in DCs derived from patients with CGD, the cross-presentation of 2 model tumor antigens was impaired. We conclude that NOX2 activity participates in the regulation of the phagosomal and endosomal pH in human DCs, and is required for efficient antigen cross-presentation.
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