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Infection and Immunity, February 2003, p. 891-903, Vol. 71, No. 2
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.2.891-903.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Phagocytosis and Intracellular Fate of Aspergillus fumigatus Conidia in Alveolar Macrophages

O. Ibrahim-Granet,1* B. Philippe,1 H. Boleti,2,{dagger} E. Boisvieux-Ulrich,3 D. Grenet,4 M. Stern,4 and J. P. Latgé1*

Unité des Aspergillus, Institut Pasteur,1 Unité de Biologie des Interactions Cellulaires, Institut Pasteur, CNRS URA 1960, 75724 Paris Cedex 15,2 Laboratoire de Cytophysiologie et Toxicologie Cellulaire, Université Paris VII, Paris,3 Service de Pneumologie, Hôpital Foch, 92 151 Suresnes Cedex, France4

Received 5 July 2002/ Returned for modification 15 August 2002/ Accepted 13 November 2002

Aspergillus fumigatus is the most prevalent airborne fungal pathogen responsible for fatal invasive aspergillosis in immunocompromised patients. Upon arrival in the lung alveolus, conidia of A. fumigatus are phagocytosed by alveolar macrophages, the major phagocytic cells of the lung. Engulfment and intracellular trafficking of A. fumigatus conidia in alveolar macrophages of two different origins, the murine cell line MH-S and human pulmonary alveolar macrophages, were analyzed by electron microscopy and immunofluorescence. Phagocytosis of A. fumigatus conidia required actin polymerization and phosphatidylinositol 3-kinase activity. Fusion of A. fumigatus phagosomes with early and late endosomes was shown by immunolabeling with specific markers for the transferrin receptor, early endosome antigen, and Rab7. Maturation of A. fumigatus phagolysosomes was monitored by using a fixable acidotropic probe, LysoTracker Red DND-99, and an anti-cathepsin D antibody. Bafilomycin A-induced inhibition of lysosomal acidification abolished the conidial killing by the macrophages. These data suggest that the maturation of A. fumigatus phagosomes results from fusion with the compartments of the endocytic pathway and that the killing of conidia depends on phagolysosome acidification. A model for the phagocytosis of A. fumigatus conidia by alveolar macrophages is proposed on the basis of these results.

* Corresponding author. Mailing address for O. Ibrahim-Granet: Unité des Aspergillus, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: 33 0 1 40 61 35 18. Fax: 33 0 1 40 61 34 19. E-mail: ogranet{at}pasteur.fr. E-mail address for J. P. Latgé: jplatge{at}pasteur.fr.

Editor: T. R. Kozel

{dagger} Present address: Molecular Virology Laboratory, Hellenic Pasteur Institute, Athens 11521, Greece.

Infection and Immunity, February 2003, p. 891-903, Vol. 71, No. 2
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.2.891-903.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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