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Infection and Immunity, May 2004, p. 2872-2878, Vol. 72, No. 5
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.5.2872-2878.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Mycobacteria Inhibit Nitric Oxide Synthase Recruitment to Phagosomes during Macrophage Infection

Barbara H. Miller,1 Rutilio A. Fratti,1 Jens F. Poschet,2 Graham S. Timmins,3 Sharon S. Master,2 Marcos Burgos,4 Michael A. Marletta,5 and Vojo Deretic2*

Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, Michigan 48109-0620,1 Department of Molecular Genetics and Microbiology,2 School of Pharmacy,3 Department of Medicine, University of New Mexico Health Sciences Center, Albuquerque, New Mexico 87131,4 Howard Hughes Medical Institute, Department of Biological Chemistry and Department of Medicinal Chemistry, University of Michigan, Ann Arbor, Michigan 48109-06065

Received 12 July 2003/ Returned for modification 6 October 2003/ Accepted 5 January 2004

Inducible nitric oxide synthase (iNOS) is a cytoplasmic protein responsible for the generation of nitric oxide (NO · ) in macrophages. In this work, we hypothesized that the intracellular localization of iNOS is significant for effective delivery of NO · to phagosomes containing ingested microorganisms. Using immunofluorescence microscopy and Western blot analysis, iNOS was shown to localize in the vicinity of phagosomes containing latex beads in stimulated macrophages. iNOS also localized to phagosomes containing Escherichia coli. The colocalization of iNOS with ingested latex beads was an actin-dependent process, since treatment with the actin microfilament disrupter cytochalasin D prevented iNOS recruitment to latex bead phagosomes. In contrast to E. coli and inert particle phagosomes, mycobacterial phagosomes did not colocalize with iNOS. This study demonstrates that (i) iNOS can be recruited to phagosomes; (ii) this recruitment is dependent on a functional actin cytoskeleton; (iii) certain microorganisms have the ability to prevent or reduce colocalization with iNOS; and (iv) spatial exclusion of iNOS may play a role in Mycobacterium tuberculosis pathogenesis.

* Corresponding author. Mailing address: Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, C915 Camino de Salud, Albuquerque, NM 87131. Phone: (505) 272-0291. Fax: (505) 272-5309. E-mail: vderetic{at}salud.unm.edu.

Editor: S. H. E. Kaufmann

Infection and Immunity, May 2004, p. 2872-2878, Vol. 72, No. 5
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.5.2872-2878.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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