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  • Review Article
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Antimicrobial reactive oxygen and nitrogen species: concepts and controversies

Nature Reviews Microbiology volume 2, pages 820–832 (2004)Cite this article

Key Points

  • The mechanisms by which phagocytes kill microorganisms are still not completely understood. Phagocyte-derived reactive oxygen species (ROS) and reactive nitrogen species (RNS) are undoubtedly important molecules in this process. Here, Ferric Fang reviews the biology of these molecules, with particular emphasis on aspects of their biology that remain controversial.

  • ROS and RNS are important molecules in host innate immunity. The author briefly reviews the diseases that occur when these molecules cannot be produced, including the roles of ROS in chronic granulomatous disease (CGD); RNS and inducible nitric oxide synthase (iNOS) promoter polymorphisms; and deficiencies in cytokine production or responses.

  • The NADPH phagocyte oxidase (phox)-mediated generation of ROS and the iNOS-mediated generation of RNS are described. The range of ROS and RNS species that are produced, and their cellular target molecules are also discussed.

  • Microorganisms can sometimes overcome the production of ROS and RNS to avoid being killed. The strategies that are used are reviewed, including evasion of ROS and RNS, suppression of ROS and RNS production, enzymic detoxification of reactive species, scavenging of the species to remove them, iron sequestration, stress responses and repairing the damage that the molecules cause.

  • Controversies in this field include the mechanisms by which nitric oxide is produced by macrophages, the roles of myeloperoxidase and xanthine oxidase, the importance of vesicular transport, the synergy between ROS and RNS, the synergy between ROS and proteases, and the generation of ROS by antibodies — all of which are discussed here.

Abstract

Phagocyte-derived reactive oxygen and nitrogen species are of crucial importance for host resistance to microbial pathogens. Decades of research have provided a detailed understanding of the regulation, generation and actions of these molecular mediators, as well as their roles in resisting infection. However, differences of opinion remain with regard to their host specificity, cell biology, sources and interactions with one another or with myeloperoxidase and granule proteases. More than a century after Metchnikoff first described phagocytosis, and more than four decades after the discovery of the burst of oxygen consumption that is associated with microbial killing, the seemingly elementary question of how phagocytes inhibit, kill and degrade microorganisms remains controversial. This review updates the reader on these concepts and the topical questions in the field.

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Figure 1: Antimicrobial mechanisms of phagocytes.
Figure 2: Reactive oxygen and nitrogen intermediate production in mammalian cells.
Figure 3: Microbial targets of reactive oxygen and nitrogen species.

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References

  1. Ambruso, D. R. et al. Human neutrophil immunodeficiency syndrome is associated with an inhibitory Rac2 mutation. Proc. Natl Acad. Sci. USA 97, 4654–4659 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Gray, G. R. et al. Neutrophil dysfunction, chronic granulomatous disease, and non-spherocytic haemolytic anaemia caused by complete deficiency of glucose-6-phosphate dehydrogenase. Lancet 2, 530–534 (1973).

    Article  CAS  PubMed  Google Scholar 

  3. Pollock, J. D. et al. Mouse model of X-linked chronic granulomatous disease, an inherited defect in phagocyte superoxide production. Nature Genet. 9, 202–209 (1995).

    Article  CAS  PubMed  Google Scholar 

  4. Jackson, S. H., Gallin, J. I. & Holland, S. M. The p47phox mouse knock-out model of chronic granulomatous disease. J. Exp. Med. 182, 751–758 (1995). Presents a model of X-linked deficiency of the NADPH phagocyte oxidase.

    Article  CAS  PubMed  Google Scholar 

  5. The, International Chronic Granulomatous Disease Cooperative Study Group. A controlled trial of interferon-γ to prevent infection in chronic granulomatous disease. N. Engl. J. Med. 324, 509–516 (1991).

  6. Jackson, S. H. et al. IFN-γ is effective in reducing infections in the mouse model of chronic granulomatous disease (CGD). J. Interferon Cytokine Res. 21, 567–573 (2001).

    Article  CAS  PubMed  Google Scholar 

  7. Muhlebach, T. J. et al. Treatment of patients with chronic granulomatous disease with recombinant human interferon-γ does not improve neutrophil oxidative metabolism, cytochrome b558 content or levels of four anti-microbial proteins. Clin. Exp. Immunol. 88, 203–206 (1992).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Levesque, M. C. et al. Nitric oxide synthase type 2 promoter polymorphisms, nitric oxide production, and disease severity in Tanzanian children with malaria. J. Infect. Dis. 180, 1994–2002 (1999). Provides the first genetic evidence of the role of iNOS in human resistance to infection.

    Article  CAS  PubMed  Google Scholar 

  9. Hobbs, M. R. et al. A new NOS2 promoter polymorphism associated with increased nitric oxide production and protection from severe malaria in Tanzanian and Kenyan children. Lancet 360, 1468–1475 (2002).

    Article  CAS  PubMed  Google Scholar 

  10. Kun, J. F. et al. Nitric oxide synthase 2 (Lambarene) (G-954C), increased nitric oxide production, and protection against malaria. J. Infect. Dis. 184, 330–336 (2001).

    Article  CAS  PubMed  Google Scholar 

  11. Lopansri, B. K. et al. Low plasma arginine concentrations in children with cerebral malaria and decreased nitric oxide production. Lancet 361, 676–678 (2003).

    Article  CAS  PubMed  Google Scholar 

  12. MacMicking, J. D. et al. Altered responses to bacterial infection and endotoxic shock in mice lacking inducible nitric oxide synthase. Cell 81, 641–650 (1995).

    Article  CAS  PubMed  Google Scholar 

  13. Nathan, C. & Shiloh, M. U. Reactive oxygen and nitrogen intermediates in the relationship between mammalian hosts and microbial pathogens. Proc. Natl Acad. Sci. USA 97, 8841–8848 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Mastroeni, P. et al. Antimicrobial actions of the NADPH phagocyte oxidase and inducible nitric oxide synthase in experimental salmonellosis. II. Effects on microbial proliferation and host survival in vivo. J. Exp. Med. 192, 237–248 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Vazquez-Torres, A., Jones-Carson, J., Mastroeni, P., Ischiropoulos, H. & Fang, F. C. Antimicrobial actions of the NADPH phagocyte oxidase and inducible nitric oxide synthase in experimental salmonellosis. I. Effects on microbial killing by activated peritoneal macrophages in vitro. J. Exp. Med. 192, 227–236 (2000). References 14 and 15 describe the sequential contributions of NADPH phagocyte oxidase and iNOS in host resistance to Salmonella infection.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Shiloh, M. U. et al. Phenotype of mice and macrophages deficient in both phagocyte oxidase and inducible nitric oxide synthase. Immunity 10, 29–38 (1999). Shows that NADPH oxidase and NO˙ synthase might compensate for one another as mice that are deficient in both develop spontaneous infections with commensal flora.

    Article  CAS  PubMed  Google Scholar 

  17. Voskuil, M. I. et al. Inhibition of respiration by nitric oxide induces a Mycobacterium tuberculosis dormancy program. J. Exp. Med. 198, 705–713 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Stenger, S., Donhauser, N., Thuring, H., Rollinghoff, M. & Bogdan, C. Reactivation of latent leishmaniasis by inhibition of inducible nitric oxide synthase. J. Exp. Med. 183, 1501–1514 (1996). Shows a clear role for NO˙ in the maintenance of latent infection.

    Article  CAS  PubMed  Google Scholar 

  19. Scanga, C. A. et al. Reactivation of latent tuberculosis: variations on the Cornell murine model. Infect. Immun. 67, 4531–4538 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Choi, H. S., Rai, P. R., Chu, H. W., Cool, C. & Chan, E. D. Analysis of nitric oxide synthase and nitrotyrosine expression in human pulmonary tuberculosis. Am. J. Respir. Crit. Care Med. 166, 178–186 (2002).

    Article  PubMed  Google Scholar 

  21. Qadoumi, M., Becker, I., Donhauser, N., Rollinghoff, M. & Bogdan, C. Expression of inducible nitric oxide synthase in skin lesions of patients with American cutaneous leishmaniasis. Infect. Immun. 70, 4638–4642 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Blos, M. et al. Organ-specific and stage-dependent control of Leishmania major infection by inducible nitric oxide synthase and phagocyte NADPH oxidase. Eur. J. Immunol. 33, 1224–1234 (2003).

    Article  CAS  PubMed  Google Scholar 

  23. Mastroeni, P. et al. Interleukin-12 is required for control of the growth of attenuated aromatic-compound-dependent salmonellae in BALB/c mice: role of γ-interferon and macrophage activation. Infect. Immun. 66, 4767–4776 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Xing, Z., Zganiacz, A. & Santosuosso, M. Role of IL-12 in macrophage activation during intracellular infection: IL-12 and mycobacteria synergistically release TNF-α and nitric oxide from macrophages via IFN-γ induction. J. Leukoc. Biol. 68, 897–902 (2000).

    CAS  PubMed  Google Scholar 

  25. de Jong, R. et al. Severe mycobacterial and Salmonella infections in interleukin-12 receptor-deficient patients. Science 280, 1435–1438 (1998).

    Article  CAS  PubMed  Google Scholar 

  26. Morahan, G. et al. A promoter polymorphism in the gene encoding interleukin-12 p40 (IL12B) is associated with mortality from cerebral malaria and with reduced nitric oxide production. Genes Immun. 3, 414–418 (2002).

    Article  CAS  PubMed  Google Scholar 

  27. Babior, B. M., Lambeth, J. D. & Nauseef, W. The neutrophil NADPH oxidase. Arch. Biochem. Biophys. 397, 342–344 (2002). An up-to-date mini-review of NADPH oxidase.

    Article  CAS  PubMed  Google Scholar 

  28. Vignais, P. V. The superoxide-generating NADPH oxidase: structural aspects and activation mechanism. Cell Mol. Life Sci. 59, 1428–1459 (2002).

    Article  CAS  PubMed  Google Scholar 

  29. Cadwallader, K. A. et al. Regulation of phosphatidylinositol 3-kinase activity and phosphatidylinositol 3,4,5-trisphosphate accumulation by neutrophil priming agents. J. Immunol. 169, 3336–3344 (2002).

    Article  CAS  PubMed  Google Scholar 

  30. Ellson, C. D. et al. PtdIns(3)P regulates the neutrophil oxidase complex by binding to the PX domain of p40(phox). Nature Cell Biol. 3, 679–682 (2001).

    Article  CAS  PubMed  Google Scholar 

  31. Cassatella, M. A. et al. Molecular basis of interferon-γ and lipopolysaccharide enhancement of phagocyte respiratory burst capability. Studies on the gene expression of several NADPH oxidase components. J. Biol. Chem. 265, 20241–20246 (1990).

    CAS  PubMed  Google Scholar 

  32. Dinauer, M. C. Regulation of neutrophil function by Rac GTPases. Curr. Opin. Hematol. 10, 8–15 (2003).

    Article  CAS  PubMed  Google Scholar 

  33. Zhao, X., Carnevale, K. A. & Cathcart, M. K. Human monocytes use Rac1, not Rac2, in the NADPH oxidase complex. J. Biol. Chem. 278, 40788–40792 (2003).

    Article  CAS  PubMed  Google Scholar 

  34. Bokoch, G. M., Quilliam, L. A., Bohl, B. P., Jesaitis, A. J. & Quinn, M. T. Inhibition of Rap1A binding to cytochrome b558 of NADPH oxidase by phosphorylation of Rap1A. Science 254, 1794–1796 (1991).

    Article  CAS  PubMed  Google Scholar 

  35. DeCoursey, T. E., Morgan, D. & Cherny, V. V. The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels. Nature 422, 531–534 (2003).

    Article  CAS  PubMed  Google Scholar 

  36. Sbarra, A. J. & Karnovsky, M. L. The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J. Biol. Chem. 234, 1355–1362 (1959).

    CAS  PubMed  Google Scholar 

  37. Hibbs, J. B. Jr. Infection and nitric oxide. J. Infect. Dis. 185 (Suppl. 1), S9–S17 (2002).

    Article  CAS  PubMed  Google Scholar 

  38. Stamler, J. S., Lamas, S. & Fang, F. C. Nitrosylation. The prototypic redox-based signaling mechanism. Cell 106, 675–683 (2001). The authors propose that thiol nitrosylation is a model for redox-based signal transduction.

    Article  CAS  PubMed  Google Scholar 

  39. Serbina, N. V., Salazar-Mather, T. P., Biron, C. A., Kuziel, W. A. & Pamer, E. G. TNF/iNOS-producing dendritic cells mediate innate immune defense against bacterial infection. Immunity 19, 59–70 (2003).

    Article  CAS  PubMed  Google Scholar 

  40. Bogdan, C., Rollinghoff, M. & Diefenbach, A. The role of nitric oxide in innate immunity. Immunol. Rev. 173, 17–26 (2000).

    Article  CAS  PubMed  Google Scholar 

  41. Benjamin, N. & Dykhuizen, R. in Nitric Oxide and Infection (ed. Fang, F. C.) 215–230 (Kluwer Academic/Plenum Publishers, New York, 1999).

    Google Scholar 

  42. Stuehr, D. J. Mammalian nitric oxide synthases. Biochim Biophys. Acta 1411, 217–230 (1999).

    Article  CAS  PubMed  Google Scholar 

  43. Stuehr, D. J. & Marletta, M. A. Mammalian nitrate biosynthesis: mouse macrophages produce nitrite and nitrate in response to Escherichia coli lipopolysaccharide. Proc. Natl Acad. Sci. USA 82, 7738–7742 (1985).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Xie, Q. W., Kashiwabara, Y. & Nathan, C. Role of transcription factor NF-κB/Rel in induction of nitric oxide synthase. J. Biol. Chem. 269, 4705–4708 (1994).

    CAS  PubMed  Google Scholar 

  45. Kamijo, R. et al. Requirement for transcription factor IRF-1 in NO synthase induction in macrophages. Science 263, 1612–1615 (1994).

    Article  CAS  PubMed  Google Scholar 

  46. Taylor, B. S. & Geller, D. A. Molecular regulation of the human inducible nitric oxide synthase (iNOS) gene. Shock 13, 413–424 (2000). Discusses the fundamental differences between human and murine iNOS promoters.

    Article  CAS  PubMed  Google Scholar 

  47. Vazquez-Torres, A. et al. Toll-like receptor 4 dependence of innate and adaptive immunity to Salmonella: importance of the Kupffer cell network. J. Immunol. 172, 6202–6208 (2004).

    Article  CAS  PubMed  Google Scholar 

  48. Kolodziejski, P. J., Musial, A., Koo, J. S. & Eissa, N. T. Ubiquitination of inducible nitric oxide synthase is required for its degradation. Proc. Natl Acad. Sci. USA 99, 12315–12320 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. El-Gayar, S., Thuring-Nahler, H., Pfeilschifter, J., Rollinghoff, M. & Bogdan, C. Translational control of inducible nitric oxide synthase by IL-13 and arginine availability in inflammatory macrophages. J. Immunol. 171, 4561–4568 (2003).

    Article  CAS  PubMed  Google Scholar 

  50. Schoedon, G. et al. Regulation of the L-arginine-dependent and tetrahydrobiopterin-dependent biosynthesis of nitric oxide in murine macrophages. Eur. J. Biochem. 213, 833–839 (1993).

    Article  CAS  PubMed  Google Scholar 

  51. Nathan, C. & Xie, Q. W. Nitric oxide synthases: roles, tolls, and controls. Cell 78, 915–918 (1994).

    Article  CAS  PubMed  Google Scholar 

  52. Gaston, B. & Stamler, J. S. in Nitric Oxide and Infection (ed. Fang, F. C.) 37–55 (Kluwer Academic/Plenum Publishers, New York, 1999).

    Google Scholar 

  53. De Groote, M. A. & Fang, F. C. in Nitric Oxide and Infection (ed. Fang, F. C.) 231–261 (Kluwer Academic/Plenum Publishers, New York, 1999).

    Google Scholar 

  54. Imlay, J. A. & Linn, S. Bimodal pattern of killing of DNA-repair-defective or anoxically grown Escherichia coli by hydrogen peroxide. J. Bacteriol. 166, 519–527 (1986). Shows that the mechanism of bacterial killing by H 2 O 2 is concentration dependent.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Imlay, J. A. & Linn, S. DNA damage and oxygen radical toxicity. Science 240, 1302–1309 (1988).

    Article  CAS  PubMed  Google Scholar 

  56. McCormick, M. L., Buettner, G. R. & Britigan, B. E. Endogenous superoxide dismutase levels regulate iron-dependent hydroxyl radical formation in Escherichia coli exposed to hydrogen peroxide. J. Bacteriol. 180, 622–625 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  57. Tamarit, J., Cabiscol, E. & Ros, J. Identification of the major oxidatively damaged proteins in Escherichia coli cells exposed to oxidative stress. J. Biol. Chem. 273, 3027–3032 (1998).

    Article  CAS  PubMed  Google Scholar 

  58. Shohet, S. B., Pitt, J., Baehner, R. L. & Poplack, D. G. Lipid peroxidation in the killing of phagocytized pneumococci. Infect. Immun. 10, 1321–1328 (1974).

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Imlay, J. A. Pathways of oxidative damage. Annu. Rev. Microbiol. 57, 395–418 (2003).

    Article  CAS  PubMed  Google Scholar 

  60. Stadler, N., Hofer, M. & Sigler, K. Mechanisms of Saccharomyces cerevisiae PMA1 H+-ATPase inactivation by Fe2+, H2O2 and Fenton reagents. Free Radic. Res. 35, 643–653 (2001).

    Article  CAS  PubMed  Google Scholar 

  61. De Groote, M. A. et al. Genetic and redox determinants of nitric oxide cytotoxicity in a Salmonella typhimurium model. Proc. Natl Acad. Sci. USA 92, 6399–6403 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Schapiro, J. M., Libby, S. J. & Fang, F. C. Inhibition of bacterial DNA replication by zinc mobilization during nitrosative stress. Proc. Natl Acad. Sci. USA 100, 8496–8501 (2003). Indicates that zinc metalloproteins that are involved in DNA replication are a crucial target of nitrogen oxides.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Pacelli, R. et al. Nitric oxide potentiates hydrogen peroxide-induced killing of Escherichia coli. J. Exp. Med. 182, 1469–1479 (1995).

    Article  CAS  PubMed  Google Scholar 

  64. Stevanin, T. M. et al. Flavohemoglobin Hmp affords inducible protection for Escherichia coli respiration, catalyzed by cytochromes bo′ or bd, from nitric oxide. J. Biol. Chem. 275, 35868–35875 (2000). Describe how NO˙ inhibits bacterial respiration, which is antagonized by a bacterial flavohaemoglobin that converts NO˙ to nitrate.

    Article  CAS  PubMed  Google Scholar 

  65. Shi, L., Jung, Y. J., Tyagi, S., Gennaro, M. L. & North, R. J. Expression of TH1-mediated immunity in mouse lungs induces a Mycobacterium tuberculosis transcription pattern characteristic of nonreplicating persistence. Proc. Natl Acad. Sci. USA 100, 241–246 (2003).

    Article  CAS  PubMed  Google Scholar 

  66. Lepoivre, M., Fieschi, F., Coves, J., Thelander, L. & Fontecave, M. Inactivation of ribonucleotide reductase by nitric oxide. Biochem. Biophys. Res. Commun. 179, 442–448 (1991).

    Article  CAS  PubMed  Google Scholar 

  67. Woodmansee, A. N. & Imlay, J. A. A mechanism by which nitric oxide accelerates the rate of oxidative DNA damage in Escherichia coli. Mol. Microbiol. 49, 11–22 (2003).

    Article  CAS  PubMed  Google Scholar 

  68. Flint, D. H., Tuminello, J. F. & Emptage, M. H. The inactivation of Fe–S clusters containing hydrolyases by superoxide. J. Biol. Chem. 268, 22369–22376 (1993).

    CAS  PubMed  Google Scholar 

  69. Keyer, K. & Imlay, J. A. Superoxide accelerates DNA damage by elevating free-iron levels. Proc. Natl Acad. Sci. USA 93, 13635–13640 (1996).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Wink, D. A. et al. DNA deaminating ability and genotoxicity of nitric oxide and its progenitors. Science 254, 1001–1003 (1991).

    Article  CAS  PubMed  Google Scholar 

  71. Burney, S., Caulfield, J. L., Niles, J. C., Wishnok, J. S. & Tannenbaum, S. R. The chemistry of DNA damage from nitric oxide and peroxynitrite. Mutat. Res. 424, 37–49 (1999).

    Article  CAS  PubMed  Google Scholar 

  72. Spek, E. J. et al. Recombinational repair is critical for survival of Escherichia coli exposed to nitric oxide. J. Bacteriol. 183, 131–138 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Evans, T. J. et al. Cytokine-treated human neutrophils contain inducible nitric oxide synthase that produces nitration of ingested bacteria. Proc. Natl Acad. Sci. USA 93, 9553–9558 (1996).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  74. Miller, R. A. & Britigan, B. E. Role of oxidants in microbial pathophysiology. Clin. Microbiol. Rev. 10, 1–18 (1997). Reviews some of the many examples of bacterial oxidative-stress resistance mechanisms that promote virulence.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  75. Celli, J. & Finlay, B. B. Bacterial avoidance of phagocytosis. Trends Microbiol. 10, 232–237 (2002).

    Article  CAS  PubMed  Google Scholar 

  76. Black, D. S. & Bliska, J. B. Identification of p130Cas as a substrate of Yersinia YopH (Yop51), a bacterial protein tyrosine phosphatase that translocates into mammalian cells and targets focal adhesions. EMBO J. 16, 2730–2744 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Rosqvist, R., Forsberg, A. & Wolf-Watz, H. Intracellular targeting of the Yersinia YopE cytotoxin in mammalian cells induces actin microfilament disruption. Infect. Immun. 59, 4562–4569 (1991).

    CAS  PubMed  PubMed Central  Google Scholar 

  78. Vazquez-Torres, A. et al. Salmonella pathogenicity island 2-dependent evasion of the phagocyte NADPH oxidase. Science 287, 1655–1658 (2000). Shows that Salmonella translocates proteins into the macrophage cytosol, which seems to interfere with vesicular transport.

    Article  CAS  PubMed  Google Scholar 

  79. Chakravortty, D., Hansen-Wester, I. & Hensel, M. Salmonella pathogenicity island 2 mediates protection of intracellular Salmonella from reactive nitrogen intermediates. J. Exp. Med. 195, 1155–1166 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Cowley, S. C., Myltseva, S. V. & Nano, F. E. Phase variation in Francisella tularensis affecting intracellular growth, lipopolysaccharide antigenicity and nitric oxide production. Mol. Microbiol. 20, 867–874 (1996).

    Article  CAS  PubMed  Google Scholar 

  81. Bliska, J. B. & Black, D. S. Inhibition of the Fc receptor-mediated oxidative burst in macrophages by the Yersinia pseudotuberculosis tyrosine phosphatase. Infect. Immun. 63, 681–685 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  82. Pearson, R. D., Symes, P., Conboy, M., Weiss, A. A. & Hewlett, E. L. Inhibition of monocyte oxidative responses by Bordetella pertussis adenylate cyclase toxin. J. Immunol. 139, 2749–2754 (1987).

    CAS  PubMed  Google Scholar 

  83. Prada, J., Malinowski, J., Muller, S., Bienzle, U. & Kremsner, P. G. Effects of Plasmodium vinckei hemozoin on the production of oxygen radicals and nitrogen oxides in murine macrophages. Am. J. Trop. Med. Hyg. 54, 620–624 (1996).

    Article  CAS  PubMed  Google Scholar 

  84. Choi, K. S. & Dumler, J. S. Early induction and late abrogation of respiratory burst in A. phagocytophilum-infected neutrophils. Ann. NY Acad. Sci. 990, 488–493 (2003).

    Article  PubMed  Google Scholar 

  85. Mott, J., Rikihisa, Y. & Tsunawaki, S. Effects of Anaplasma phagocytophila on NADPH oxidase components in human neutrophils and HL-60 cells. Infect. Immun. 70, 1359–1366 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Carlyon, J. A., Chan, W. T., Galan, J., Roos, D. & Fikrig, E. Repression of rac2 mRNA expression by Anaplasma phagocytophila is essential to the inhibition of superoxide production and bacterial proliferation. J. Immunol. 169, 7009–7018 (2002).

    Article  CAS  PubMed  Google Scholar 

  87. Saura, M. et al. An antiviral mechanism of nitric oxide: inhibition of a viral protease. Immunity 10, 21–28 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Colasanti, M., Persichini, T., Venturini, G. & Ascenzi, P. S-nitrosylation of viral proteins: molecular bases for antiviral effect of nitric oxide. IUBMB Life 48, 25–31 (1999).

    Article  CAS  PubMed  Google Scholar 

  89. Cao, W., Bao, C. & Lowenstein, C. J. Inducible nitric oxide synthase expression inhibition by adenovirus E1A. Proc. Natl Acad. Sci. USA 100, 7773–7778 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  90. Seyler, R. W. Jr, Olson, J. W. & Maier, R. J. Superoxide dismutase-deficient mutants of Helicobacter pylori are hypersensitive to oxidative stress and defective in host colonization. Infect. Immun. 69, 4034–4040 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  91. Fang, F. C. et al. Virulent Salmonella typhimurium has two periplasmic Cu, Zn-superoxide dismutases. Proc. Natl Acad. Sci. USA 96, 7502–7507 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Wilson, T. M., de Lisle, G. W. & Collins, D. M. Effect of inhA and katG on isoniazid resistance and virulence of Mycobacterium bovis. Mol. Microbiol. 15, 1009–1015 (1995).

    Article  CAS  PubMed  Google Scholar 

  93. Bishai, W. R., Howard, N. S., Winkelstein, J. A. & Smith, H. O. Characterization and virulence analysis of catalase mutants of Haemophilus influenzae. Infect. Immun. 62, 4855–4860 (1994).

    CAS  PubMed  PubMed Central  Google Scholar 

  94. Poole, R. K. & Hughes, M. N. New functions for the ancient globin family: bacterial responses to nitric oxide and nitrosative stress. Mol. Microbiol. 36, 775–783 (2000).

    Article  CAS  PubMed  Google Scholar 

  95. Pathania, R., Navani, N. K., Gardner, A. M., Gardner, P. R. & Dikshit, K. L. Nitric oxide scavenging and detoxification by the Mycobacterium tuberculosis haemoglobin, HbN in Escherichia coli. Mol. Microbiol. 45, 1303–1314 (2002).

    Article  CAS  PubMed  Google Scholar 

  96. Gardner, A. M., Helmick, R. A. & Gardner, P. R. Flavorubredoxin, an inducible catalyst for nitric oxide reduction and detoxification in Escherichia coli. J. Biol. Chem. 277, 8172–8177 (2002).

    Article  CAS  PubMed  Google Scholar 

  97. Liu, L. et al. A metabolic enzyme for S-nitrosothiol conserved from bacteria to humans. Nature 410, 490–494 (2001).

    Article  CAS  PubMed  Google Scholar 

  98. Bryk, R., Griffin, P. & Nathan, C. Peroxynitrite reductase activity of bacterial peroxiredoxins. Nature 407, 211–215 (2000).

    Article  CAS  PubMed  Google Scholar 

  99. de Jesus-Berrios, M. et al. Enzymes that counteract nitrosative stress promote fungal virulence. Curr. Biol. 13, 1963–1968 (2003).

    Article  CAS  PubMed  Google Scholar 

  100. Newton, G. L. et al. Distribution of thiols in microorganisms: mycothiol is a major thiol in most actinomycetes. J. Bacteriol. 178, 1990–1995 (1996).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  101. Thomson, L., Denicola, A. & Radi, R. The trypanothione–thiol system in Trypanosoma cruzi as a key antioxidant mechanism against peroxynitrite-mediated cytotoxicity. Arch. Biochem. Biophys. 412, 55–64 (2003).

    Article  CAS  PubMed  Google Scholar 

  102. Brenot, A., King, K. Y., Janowiak, B., Griffith, O. & Caparon, M. G. Contribution of glutathione peroxidase to the virulence of Streptococcus pyogenes. Infect. Immun. 72, 408–413 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  103. St John, G. et al. Peptide methionine sulfoxide reductase from Escherichia coli and Mycobacterium tuberculosis protects bacteria against oxidative damage from reactive nitrogen intermediates. Proc. Natl Acad. Sci. USA 98, 9901–9906 (2001).

    Article  CAS  PubMed  Google Scholar 

  104. De Groote, M. A., Testerman, T., Xu, Y., Stauffer, G. & Fang, F. C. Homocysteine antagonism of nitric oxide-related cytostasis in Salmonella typhimurium. Science 272, 414–417 (1996). Shows that homocysteine antagonizes the actions of NO˙ in bacteria as well as in blood vessels.

    Article  CAS  PubMed  Google Scholar 

  105. van Guldener, C. & Stehouwer, C. D. Hyperhomocysteinemia, vascular pathology, and endothelial dysfunction. Semin. Thromb. Hemost. 26, 281–289 (2000).

    Article  CAS  PubMed  Google Scholar 

  106. Wang, Y., Aisen, P. & Casadevall, A. Cryptococcus neoformans melanin and virulence: mechanism of action. Infect. Immun. 63, 3131–3136 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  107. Chaturvedi, V., Wong, B. & Newman, S. L. Oxidative killing of Cryptococcus neoformans by human neutrophils. Evidence that fungal mannitol protects by scavenging reactive oxygen intermediates. J. Immunol. 156, 3836–3840 (1996).

    CAS  PubMed  Google Scholar 

  108. Simpson, J. A., Smith, S. E. & Dean, R. T. Scavenging by alginate of free radicals released by macrophages. Free Radic. Biol. Med. 6, 347–353 (1989).

    Article  CAS  PubMed  Google Scholar 

  109. Andrews, S. C. Iron storage in bacteria. Adv. Microb. Physiol. 40, 281–351 (1998).

    Article  CAS  PubMed  Google Scholar 

  110. Almiron, M., Link, A. J., Furlong, D. & Kolter, R. A novel DNA-binding protein with regulatory and protective roles in starved Escherichia coli. Genes Dev. 6, 2646–2654 (1992).

    Article  CAS  PubMed  Google Scholar 

  111. Hantke, K. Iron and metal regulation in bacteria. Curr. Opin. Microbiol. 4, 172–177 (2001).

    Article  CAS  PubMed  Google Scholar 

  112. Chen, C. Y. & Morse, S. A. Neisseria gonorrhoeae bacterioferritin: structural heterogeneity, involvement in iron storage and protection against oxidative stress. Microbiology 145, 2967–2975 (1999).

    Article  CAS  PubMed  Google Scholar 

  113. Wai, S. N., Nakayama, K., Umene, K., Moriya, T. & Amako, K. Construction of a ferritin-deficient mutant of Campylobacter jejuni: contribution of ferritin to iron storage and protection against oxidative stress. Mol. Microbiol. 20, 1127–1134 (1996).

    Article  CAS  PubMed  Google Scholar 

  114. Halsey, T. A., Vazquez-Torres, A., Gravdahl, D. J., Fang, F. C. & Libby, S. J. The ferritin-like Dps protein is required for Salmonella enterica serovar typhimurium oxidative stress resistance and virulence. Infect. Immun. 72, 1155–1158 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  115. D'Autreaux, B., Touati, D., Bersch, B., Latour, J. M. & Michaud-Soret, I. Direct inhibition by nitric oxide of the transcriptional ferric uptake regulation protein via nitrosylation of the iron. Proc. Natl Acad. Sci. USA 99, 16619–16624 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  116. Lobysheva, I. I., Stupakova, M. V., Mikoyan, V. D., Vasilieva, S. V. & Vanin, A. F. Induction of the SOS DNA repair response in Escherichia coli by nitric oxide donating agents: dinitrosyl iron complexes with thiol-containing ligands and S-nitrosothiols. FEBS Lett. 454, 177–180 (1999).

    Article  CAS  PubMed  Google Scholar 

  117. Christman, M. F., Morgan, R. W., Jacobson, F. S. & Ames, B. N. Positive control of a regulon for defenses against oxidative stress and some heat-shock proteins in Salmonella typhimurium. Cell 41, 753–762 (1985).

    Article  CAS  PubMed  Google Scholar 

  118. Greenberg, J. T., Monach, P., Chou, J. H., Josephy, P. D. & Demple, B. Positive control of a global antioxidant defense regulon activated by superoxide-generating agents in Escherichia coli. Proc. Natl Acad. Sci. USA 87, 6181–6185 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  119. Zheng, M., Aslund, F. & Storz, G. Activation of the OxyR transcription factor by reversible disulfide bond formation. Science 279, 1718–1721 (1998).

    Article  CAS  PubMed  Google Scholar 

  120. Paget, M. S., Kang, J. G., Roe, J. H. & Buttner, M. J. σR, an RNA polymerase σ-factor that modulates expression of the thioredoxin system in response to oxidative stress in Streptomyces coelicolor A3(2). EMBO J. 17, 5776–5782 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  121. Helmann, J. D. et al. The global transcriptional response of Bacillus subtilis to peroxide stress is coordinated by three transcription factors. J. Bacteriol. 185, 243–253 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  122. Boylan, J. A., Posey, J. E. & Gherardini, F. C. Borrelia oxidative stress response regulator, BosR: a distinctive Zn-dependent transcriptional activator. Proc. Natl Acad. Sci. USA 100, 11684–11689 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  123. Sak, B. D., Eisenstark, A. & Touati, D. Exonuclease III and the catalase hydroperoxidase II in Escherichia coli are both regulated by the katF gene product. Proc. Natl Acad. Sci. USA 86, 3271–3275 (1989).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  124. Mukhopadhyay, P., Zheng, M., Bedzyk, L. A., LaRossa, R. A. & Storz, G. Prominent roles of the NorR and Fur regulators in the Escherichia coli transcriptional response to reactive nitrogen species. Proc. Natl Acad. Sci. USA 101, 745–750 (2004). The authors argue that, despite some overlap, there seem to be important differences in the microbial transcriptional responses to oxidative and nitrosative stress.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  125. Cruz-Ramos, H. et al. NO sensing by FNR: regulation of the Escherichia coli NO-detoxifying flavohaemoglobin, Hmp. EMBO J. 21, 3235–3244 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  126. Zumft, W. G. Nitric oxide signaling and NO dependent transcriptional control in bacterial denitrification by members of the FNR-CRP regulator family. J. Mol. Microbiol. Biotechnol. 4, 277–286 (2002).

    CAS  PubMed  Google Scholar 

  127. Suvarnapunya, A. E., Lagasse, H. A. & Stein, M. A. The role of DNA base excision repair in the pathogenesis of Salmonella enterica serovar typhimurium. Mol. Microbiol. 48, 549–559 (2003).

    Article  CAS  PubMed  Google Scholar 

  128. Darwin, K. H., Ehrt, S., Gutierrez-Ramos, J. C., Weich, N. & Nathan, C. F. The proteasome of Mycobacterium tuberculosis is required for resistance to nitric oxide. Science 302, 1963–1966 (2003).

    Article  CAS  PubMed  Google Scholar 

  129. Buchmeier, N. A., Lipps, C. J., So, M. Y. & Heffron, F. Recombination-deficient mutants of Salmonella typhimurium are avirulent and sensitive to the oxidative burst of macrophages. Mol. Microbiol. 7, 933–936 (1993).

    Article  CAS  PubMed  Google Scholar 

  130. O'Rourke, E. J. et al. Pathogen DNA as target for host-generated oxidative stress: role for repair of bacterial DNA damage in Helicobacter pylori colonization. Proc. Natl Acad. Sci. USA 100, 2789–2794 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  131. Buchmeier, N. A. et al. DNA repair is more important than catalase for Salmonella virulence in mice. J. Clin. Invest. 95, 1047–1053 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  132. Brawn, M. K. & Fridovich, I. Increased superoxide radical production evokes inducible DNA repair in Escherichia coli. J. Biol. Chem. 260, 922–925 (1985).

    CAS  PubMed  Google Scholar 

  133. Zheng, M. et al. DNA microarray-mediated transcriptional profiling of the Escherichia coli response to hydrogen peroxide. J. Bacteriol. 183, 4562–4570 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  134. Gerschman, R., Gilbert, D. L., Nye, S. W., Dwyer, P. & Fenn, W. O. Oxygen poisoning and X-irradiation: a mechanism in common. Science 119, 623–626 (1954).

    Article  CAS  PubMed  Google Scholar 

  135. Schneemann, M. et al. Nitric oxide synthase is not a constituent of the antimicrobial armature of human mononuclear phagocytes. J. Infect. Dis. 167, 1358–1363 (1993).

    Article  CAS  PubMed  Google Scholar 

  136. Weinberg, J. B. Nitric oxide production and nitric oxide synthase type 2 expression by human mononuclear phagocytes: a review. Mol. Med. 4, 557–591 (1998). Reviews compelling evidence for NO˙ production by human macrophages.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  137. Wheeler, M. A. et al. Bacterial infection induces nitric oxide synthase in human neutrophils. J. Clin. Invest. 99, 110–116 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  138. Schneemann, M. & Schoedon, G. Species differences in macrophage NO production are important. Nature Immunol. 3, 102 (2002).

    Article  CAS  Google Scholar 

  139. Fang, F. C. & Vazquez-Torres, A. Nitric oxide production by human macrophages: there's NO doubt about it. Am. J. Physiol. Lung Cell Mol. Physiol. 282, L941–L943 (2002).

    Article  CAS  PubMed  Google Scholar 

  140. Nicholson, S. et al. Inducible nitric oxide synthase in pulmonary alveolar macrophages from patients with tuberculosis. J. Exp. Med. 183, 2293–2302 (1996).

    Article  CAS  PubMed  Google Scholar 

  141. Annane, D. et al. Compartmentalised inducible nitric-oxide synthase activity in septic shock. Lancet 355, 1143–1148 (2000).

    Article  CAS  PubMed  Google Scholar 

  142. Pham, T. N. et al. Elevated serum nitric oxide levels in patients with inflammatory arthritis associated with co-expression of inducible nitric oxide synthase and protein kinase C-η in peripheral blood monocyte-derived macrophages. J. Rheumatol. 30, 2529–2534 (2003).

    CAS  PubMed  Google Scholar 

  143. Nauseef, W. M. & Malech, H. L. Analysis of the peptide subunits of human neutrophil myeloperoxidase. Blood 67, 1504–1507 (1986).

    CAS  PubMed  Google Scholar 

  144. Daugherty, A., Dunn, J. L., Rateri, D. L. & Heinecke, J. W. Myeloperoxidase, a catalyst for lipoprotein oxidation, is expressed in human atherosclerotic lesions. J. Clin. Invest. 94, 437–444 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  145. Foote, C. S., Goyne, T. E. & Lehrer, R. I. Assessment of chlorination by human neutrophils. Nature 301, 715–716 (1983).

    Article  CAS  PubMed  Google Scholar 

  146. Eiserich, J. P. et al. Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils. Nature 391, 393–397 (1998).

    Article  CAS  PubMed  Google Scholar 

  147. Klebanoff, S. J. Iodination of bacteria: a bactericidal mechanism. J. Exp. Med. 126, 1063–1078 (1967).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  148. Rosen, H. & Klebanoff, S. J. Bactericidal activity of a superoxide anion-generating system. A model for the polymorphonuclear leukocyte. J. Exp. Med. 149, 27–39 (1979).

    Article  CAS  PubMed  Google Scholar 

  149. Rosen, H., Orman, J., Rakita, R. M., Michel, B. R. & VanDevanter, D. R. Loss of DNA–membrane interactions and cessation of DNA synthesis in myeloperoxidase-treated Escherichia coli. Proc. Natl Acad. Sci. USA 87, 10048–10052 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  150. Thomas, E. L. & Aune, T. M. Oxidation of Escherichia coli sulfhydryl components by the peroxidase–hydrogen peroxide–iodide antimicrobial system. Antimicrob. Agents Chemother. 13, 1006–1010 (1978).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  151. Rakita, R. M., Michel, B. R. & Rosen, H. Myeloperoxidase-mediated inhibition of microbial respiration: damage to Escherichia coli ubiquinol oxidase. Biochemistry 28, 3031–3036 (1989).

    Article  CAS  PubMed  Google Scholar 

  152. Rosen, H., Crowley, J. R. & Heinecke, J. W. Human neutrophils use the myeloperoxidase–hydrogen peroxide–chloride system to chlorinate but not nitrate bacterial proteins during phagocytosis. J. Biol. Chem. 277, 30463–30468 (2002).

    Article  CAS  PubMed  Google Scholar 

  153. Chapman, A. L., Hampton, M. B., Senthilmohan, R., Winterbourn, C. C. & Kettle, A. J. Chlorination of bacterial and neutrophil proteins during phagocytosis and killing of Staphylococcus aureus. J. Biol. Chem. 277, 9757–9762 (2002).

    Article  CAS  PubMed  Google Scholar 

  154. Saito, M. et al. H2O2-nonproducing Streptococcus pyogenes strains: survival in stationary phase and virulence in chronic granulomatous disease. Microbiology 147, 2469–2477 (2001).

    Article  CAS  PubMed  Google Scholar 

  155. Kottilil, S., Malech, H. L., Gill, V. J. & Holland, S. M. Infections with Haemophilus species in chronic granulomatous disease: insights into the interaction of bacterial catalase and H2O2 production. Clin. Immunol. 106, 226–230 (2003).

    Article  CAS  PubMed  Google Scholar 

  156. Parry, M. F. et al. Myeloperoxidase deficiency: prevalence and clinical significance. Ann. Intern. Med. 95, 293–301 (1981). Shows that humans with MPO deficiency have little or no immunocompromise, in contrast to individuals with CGD.

    Article  CAS  PubMed  Google Scholar 

  157. Aratani, Y. et al. Severe impairment in early host defense against Candida albicans in mice deficient in myeloperoxidase. Infect. Immun. 67, 1828–1836 (1999). Presents a description of an MPO-deficient mouse model.

    CAS  PubMed  PubMed Central  Google Scholar 

  158. Brennan, M. L. et al. Increased atherosclerosis in myeloperoxidase-deficient mice. J. Clin. Invest. 107, 419–430 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  159. Eiserich, J. P. et al. Myeloperoxidase, a leukocyte-derived vascular NO oxidase. Science 296, 2391–2394 (2002).

    Article  CAS  PubMed  Google Scholar 

  160. Brennan, M. L. et al. Prognostic value of myeloperoxidase in patients with chest pain. N. Engl. J. Med. 349, 1595–1604 (2003).

    Article  CAS  PubMed  Google Scholar 

  161. Vorbach, C., Harrison, R. & Capecchi, M. R. Xanthine oxidoreductase is central to the evolution and function of the innate immune system. Trends Immunol. 24, 512–517 (2003).

    Article  CAS  PubMed  Google Scholar 

  162. Takao, S. et al. Role of reactive oxygen metabolites in murine peritoneal macrophage phagocytosis and phagocytic killing. Am. J. Physiol. 271, C1278–C1284 (1996).

    Article  CAS  PubMed  Google Scholar 

  163. Umezawa, K. et al. Induction of nitric oxide synthesis and xanthine oxidase and their roles in the antimicrobial mechanism against Salmonella typhimurium infection in mice. Infect. Immun. 65, 2932–2940 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  164. Segal, B. H. et al. Xanthine oxidase contributes to host defense against Burkholderia cepacia in the p47phox−/− mouse model of chronic granulomatous disease. Infect. Immun. 68, 2374–2378 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  165. Simmonds, H. A., Goday, A. & Morris, G. S. Superoxide radicals, immunodeficiency and xanthine oxidase activity: man is not a mouse! Clin. Sci. (Lond) 68, 561–565 (1985).

    Article  CAS  Google Scholar 

  166. Aratani, Y. et al. Relative contributions of myeloperoxidase and NADPH-oxidase to the early host defense against pulmonary infections with Candida albicans and Aspergillus fumigatus. Med. Mycol. 40, 557–563 (2002).

    Article  CAS  PubMed  Google Scholar 

  167. Miyakawa, H. et al. Effects of inducible nitric oxide synthase and xanthine oxidase inhibitors on SEB-induced interstitial pneumonia in mice. Eur. Respir. J. 19, 447–457 (2002).

    Article  CAS  PubMed  Google Scholar 

  168. Moorhouse, P. C., Grootveld, M., Halliwell, B., Quinlan, J. G. & Gutteridge, J. M. Allopurinol and oxypurinol are hydroxyl radical scavengers. FEBS Lett. 213, 23–28 (1987).

    Article  CAS  PubMed  Google Scholar 

  169. Frayha, R. A., Salti, I. S., Arnaout, A., Khatchadurian, A. & Uthman, S. M. Hereditary xanthinuria: report on three patients and short review of the literature. Nephron 19, 328–332 (1977).

    Article  CAS  PubMed  Google Scholar 

  170. Vorbach, C., Scriven, A. & Capecchi, M. R. The housekeeping gene xanthine oxidoreductase is necessary for milk fat droplet enveloping and secretion: gene sharing in the lactating mammary gland. Genes Dev. 16, 3223–3235 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  171. Wang, J. et al. Serum xanthine oxidase: origin, regulation, and contribution to control of trypanosome parasitemia. Antioxid. Redox Signal. 4, 161–178 (2002).

    Article  CAS  PubMed  Google Scholar 

  172. Sengelov, H., Nielsen, M. H. & Borregaard, N. Separation of human neutrophil plasma membrane from intracellular vesicles containing alkaline phosphatase and NADPH oxidase activity by free flow electrophoresis. J. Biol. Chem. 267, 14912–14917 (1992).

    CAS  PubMed  Google Scholar 

  173. Badwey, J. A. et al. Comparative aspects of oxidative metabolism of neutrophils from human blood and guinea pig peritonea: magnitude of the respiratory burst, dependence upon stimulating agents, and localization of the oxidases. J. Cell. Physiol. 105, 541–545 (1980).

    Article  CAS  PubMed  Google Scholar 

  174. Ohno, Y., Hirai, K., Kanoh, T., Uchino, H. & Ogawa, K. Subcellular localization of H2O2 production in human neutrophils stimulated with particles and an effect of cytochalasin-B on the cells. Blood 60, 253–260 (1982).

    CAS  PubMed  Google Scholar 

  175. Heyworth, P. G. et al. Neutrophil nicotinamide adenine dinucleotide phosphate oxidase assembly. Translocation of p47-phox and p67-phox requires interaction between p47-phox and cytochrome b558 . J. Clin. Invest. 87, 352–356 (1991).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  176. Allen, L. A. et al. Transient association of the nicotinamide adenine dinucleotide phosphate oxidase subunits p47phox and p67phox with phagosomes in neutrophils from patients with X-linked chronic granulomatous disease. Blood 93, 3521–3530 (1999).

    CAS  PubMed  Google Scholar 

  177. Kobayashi, T., Robinson, J. M. & Seguchi, H. Identification of intracellular sites of superoxide production in stimulated neutrophils. J. Cell Sci. 111, 81–91 (1998). Shows that active NADPH oxidase might assemble in vesicles that subsequently move to the plasma membrane or phagosomal compartments.

    CAS  PubMed  Google Scholar 

  178. Seguchi, H. & Kobayashi, T. Study of NADPH oxidase-activated sites in human neutrophils. J. Electron Microsc. (Tokyo) 51, 87–91 (2002).

    Article  CAS  Google Scholar 

  179. Lang, M. L. & Kerr, M. A. Neutrophil NADPH oxidase does not assemble on macropinocytic vacuole membranes. Immunol. Lett. 72, 1–6 (2000).

    Article  CAS  PubMed  Google Scholar 

  180. Kobayashi, T. et al. A simple approach for the analysis of intracellular movement of oxidant-producing intracellular compartments in living human neutrophils. Histochem. Cell Biol. 113, 251–257 (2000).

    Article  CAS  PubMed  Google Scholar 

  181. Johansson, A. et al. Different subcellular localization of cytochrome b and the dormant NADPH-oxidase in neutrophils and macrophages: effect on the production of reactive oxygen species during phagocytosis. Cell. Immunol. 161, 61–71 (1995).

    Article  CAS  PubMed  Google Scholar 

  182. Badwey, J. A. et al. Comparative biochemical and cytochemical studies on superoxide and peroxide in mouse macrophages. J. Cell. Physiol. 115, 208–216 (1983).

    Article  CAS  PubMed  Google Scholar 

  183. Calafat, J. et al. Evidence for small intracellular vesicles in human blood phagocytes containing cytochrome b558 and the adhesion molecule CD11b/CD18. Blood 81, 3122–3129 (1993).

    CAS  PubMed  Google Scholar 

  184. Teixeira, C. F., Azevedo, N. L., Carvalho, T. M., Fuentes, J. & Nagao, P. E. Cytochemical study of Streptococcus agalactiae and macrophage interaction. Microsc. Res. Tech. 54, 254–259 (2001).

    Article  CAS  PubMed  Google Scholar 

  185. Vazquez-Torres, A., Fantuzzi, G., Edwards, C. K., Dinarello, C. A. & Fang, F. C. Defective localization of the NADPH phagocyte oxidase to Salmonella-containing phagosomes in tumor necrosis factor p55 receptor-deficient macrophages. Proc. Natl Acad. Sci. USA 98, 2561–2565 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  186. Vodovotz, Y., Russell, D., Xie, Q. W., Bogdan, C. & Nathan, C. Vesicle membrane association of nitric oxide synthase in primary mouse macrophages. J. Immunol. 154, 2914–2925 (1995).

    CAS  PubMed  Google Scholar 

  187. Fang, F. & Vazquez-Torres, A. Salmonella selectively stops traffic. Trends Microbiol. 10, 391–392 (2002).

    Article  CAS  PubMed  Google Scholar 

  188. Webb, J. L., Harvey, M. W., Holden, D. W. & Evans, T. J. Macrophage nitric oxide synthase associates with cortical actin but is not recruited to phagosomes. Infect. Immun. 69, 6391–6400 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  189. Brunelli, L., Crow, J. P. & Beckman, J. S. The comparative toxicity of nitric oxide and peroxynitrite to Escherichia coli. Arch. Biochem. Biophys. 316, 327–334 (1995).

    Article  CAS  PubMed  Google Scholar 

  190. Hickman-Davis, J., Gibbs-Erwin, J., Lindsey, J. R. & Matalon, S. Surfactant protein A mediates mycoplasmacidal activity of alveolar macrophages by production of peroxynitrite. Proc. Natl Acad. Sci. USA 96, 4953–4958 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  191. Vazquez-Torres, A., Jones-Carson, J. & Balish, E. Peroxynitrite contributes to the candidacidal activity of nitric oxide-producing macrophages. Infect. Immun. 64, 3127–3133 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  192. Darrah, P. A., Hondalus, M. K., Chen, Q., Ischiropoulos, H. & Mosser, D. M. Cooperation between reactive oxygen and nitrogen intermediates in killing of Rhodococcus equi by activated macrophages. Infect. Immun. 68, 3587–3593 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  193. De Groote, M. A. et al. Periplasmic superoxide dismutase protects Salmonella from products of phagocyte NADPH-oxidase and nitric oxide synthase. Proc. Natl Acad. Sci. USA 94, 13997–14001 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  194. Reeves, E. P. et al. Killing activity of neutrophils is mediated through activation of proteases by K+ flux. Nature 416, 291–297 (2002). This report proposes a new mechanism for the synergy between NADPH phagocyte oxidase and neutrophil granule proteases.

    Article  CAS  PubMed  Google Scholar 

  195. Tkalcevic, J. et al. Impaired immunity and enhanced resistance to endotoxin in the absence of neutrophil elastase and cathepsin G. Immunity 12, 201–210 (2000).

    Article  CAS  PubMed  Google Scholar 

  196. Ahluwalia, J. et al. The large-conductance Ca2+-activated K+ channel is essential for innate immunity. Nature 427, 853–858 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  197. Roos, D. & Winterbourn, C. C. Immunology. Lethal weapons. Science 296, 669–671 (2002).

    Article  CAS  PubMed  Google Scholar 

  198. Staudinger, B. J., Oberdoerster, M. A., Lewis, P. J. & Rosen, H. mRNA expression profiles for Escherichia coli ingested by normal and phagocyte oxidase-deficient human neutrophils. J. Clin. Invest. 110, 1151–1163 (2002). Presents evidence that the bacteria in neutrophils experience oxidative stress and that their ability to respond is crucial for their intracellular survival.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  199. Belaaouaj, A. et al. Mice lacking neutrophil elastase reveal impaired host defense against Gram-negative bacterial sepsis. Nature Med. 4, 615–618 (1998).

    Article  CAS  PubMed  Google Scholar 

  200. MacIvor, D. M. et al. Normal neutrophil function in cathepsin G-deficient mice. Blood 94, 4282–4293 (1999).

    CAS  PubMed  Google Scholar 

  201. Murray, H. W. & Cartelli, D. M. Killing of intracellular Leishmania donovani by human mononuclear phagocytes. Evidence for oxygen-dependent and -independent leishmanicidal activity. J. Clin. Invest. 72, 32–44 (1983).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  202. Mandell, G. L. & Hook, E. W. Leukocyte bactericidal activity in chronic granulomatous disease: correlation of bacterial hydrogen peroxide production and susceptibility to intracellular killing. J. Bacteriol. 100, 531–532 (1969).

    CAS  PubMed  PubMed Central  Google Scholar 

  203. Kim, Y. M., Hong, S. J., Billiar, T. R. & Simmons, R. L. Counterprotective effect of erythrocytes in experimental bacterial peritonitis is due to scavenging of nitric oxide and reactive oxygen intermediates. Infect. Immun. 64, 3074–3080 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  204. Speert, D. P., Bond, M., Woodman, R. C. & Curnutte, J. T. Infection with Pseudomonas cepacia in chronic granulomatous disease: role of nonoxidative killing by neutrophils in host defense. J. Infect. Dis. 170, 1524–1531 (1994).

    Article  CAS  PubMed  Google Scholar 

  205. Dinauer, M. C., Gifford, M. A., Pech, N., Li, L. L. & Emshwiller, P. Variable correction of host defense following gene transfer and bone marrow transplantation in murine X-linked chronic granulomatous disease. Blood 97, 3738–3745 (2001).

    Article  CAS  PubMed  Google Scholar 

  206. Harrison, R. E., Touret, N. & Grinstein, S. Microbial killing: oxidants, proteases and ions. Curr. Biol. 12, R357–R359 (2002).

    Article  CAS  PubMed  Google Scholar 

  207. Wentworth, P. Jr et al. Evidence for antibody-catalyzed ozone formation in bacterial killing and inflammation. Science 298, 2195–2199 (2002). Suggests that ozone is an antimicrobial mediator that is formed by antibodies.

    Article  CAS  PubMed  Google Scholar 

  208. Takeuchi, K. & Ibusuki, T. Quantitative determination of aqueous-phase ozone by chemiluminescence using indigo-5,5′-disulfonate. Anal. Chem. 61, 619–623 (1989).

    Article  CAS  PubMed  Google Scholar 

  209. Kettle, A. J., Clark, B. M. & Winterbourn, C. C. Superoxide converts indigo carmine to isatin sulfonic acid: implications for the hypothesis that neutrophils produce ozone. J. Biol. Chem. 279, 18521–18525 (2004).

    Article  CAS  PubMed  Google Scholar 

  210. Parren, P. W., Leusen, J. H. & van de Winkel, J. G. Antibody-catalyzed water oxidation: state-of-the-art immunity or ancient history? Trends Immunol. 24, 467–469 (2003).

    Article  CAS  PubMed  Google Scholar 

  211. Forman, H. J. & Torres, M. Reactive oxygen species and cell signaling: respiratory burst in macrophage signaling. Am. J. Respir. Crit. Care Med. 166, S4–S8 (2002).

    Article  PubMed  Google Scholar 

  212. Kim, S. & Ponka, P. Role of nitric oxide in cellular iron metabolism. Biometals 16, 125–135 (2003).

    Article  CAS  PubMed  Google Scholar 

  213. Nathan, C. Specificity of a third kind: reactive oxygen and nitrogen intermediates in cell signaling. J. Clin. Invest. 111, 769–778 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  214. Ehrt, S. et al. Reprogramming of the macrophage transcriptome in response to interferon-γ and Mycobacterium tuberculosis: signaling roles of nitric oxide synthase-2 and phagocyte oxidase. J. Exp. Med. 194, 1123–1140 (2001). Describes the profound effects of NADPH oxidase and iNOS on gene expression.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  215. Kobayashi, S. D. et al. Gene expression profiling provides insight into the pathophysiology of chronic granulomatous disease. J. Immunol. 172, 636–643 (2004).

    Article  CAS  PubMed  Google Scholar 

  216. Cooper, A. M., Adams, L. B., Dalton, D. K., Appelberg, R. & Ehlers, S. IFN-γ and NO in mycobacterial disease: new jobs for old hands. Trends Microbiol. 10, 221–226 (2002).

    Article  CAS  PubMed  Google Scholar 

  217. Alam, M. S. et al. Role of nitric oxide in host defense in murine salmonellosis as a function of its antibacterial and antiapoptotic activities. Infect. Immun. 70, 3130–3142 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  218. Sadikot, R. T. et al. p47phox deficiency impairs NF-κB activation and host defense in Pseudomonas pneumonia. J. Immunol. 172, 1801–1808 (2004).

    Article  CAS  PubMed  Google Scholar 

  219. Huang, J. et al. The quantity of nitric oxide released by macrophages regulates Chlamydia-induced disease. Proc. Natl Acad. Sci. USA 99, 3914–3919 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  220. van der Veen, R. C. et al. Superoxide prevents nitric oxide-mediated suppression of helper T lymphocytes: decreased autoimmune encephalomyelitis in nicotinamide adenine dinucleotide phosphate oxidase knockout mice. J. Immunol. 164, 5177–5183 (2000).

    Article  CAS  PubMed  Google Scholar 

  221. Li, H. & Forstermann, U. Nitric oxide in the pathogenesis of vascular disease. J. Pathol. 190, 244–254 (2000).

    Article  CAS  PubMed  Google Scholar 

  222. Fang, F. C. Nitric Oxide and Infection 517 (Kluwer Academic/Plenum Publishers, New York, 1999). A comprehensive examination of the many roles of NO˙ in infection.

    Google Scholar 

  223. Petros, A. et al. Effects of a nitric oxide synthase inhibitor in humans with septic shock. Cardiovasc. Res. 28, 34–39 (1994).

    Article  CAS  PubMed  Google Scholar 

  224. Cobb, J. P. Use of nitric oxide synthase inhibitors to treat septic shock: the light has changed from yellow to red. Crit. Care Med. 27, 855–856 (1999). Argues that although initial clinical studies of NO˙ inhibition in sepsis have been disappointing, selective inhibitors await more extensive evaluation.

    Article  CAS  PubMed  Google Scholar 

  225. Akaike, T. et al. Pathogenesis of influenza virus-induced pneumonia: involvement of both nitric oxide and oxygen radicals. Proc. Natl Acad. Sci. USA 93, 2448–2453 (1996). Shows that the inhibition of ROS/RNS-mediated tissue damage can reduce mortality in certain infections.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  226. Adler, H. et al. Suppression of herpes simplex virus type 1 (HSV-1)-induced pneumonia in mice by inhibition of inducible nitric oxide synthase (iNOS, NOS2). J. Exp. Med. 185, 1533–1540 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  227. Davis, I. C. et al. Elevated generation of reactive oxygen/nitrogen species in hantavirus cardiopulmonary syndrome. J. Virol. 76, 8347–8259 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  228. Fujii, S., Akaike, T. & Maeda, H. Role of nitric oxide in pathogenesis of herpes simplex virus encephalitis in rats. Virology 256, 203–212 (1999).

    Article  CAS  PubMed  Google Scholar 

  229. Suntres, Z. E., Omri, A. & Shek, P. N. Pseudomonas aeruginosa-induced lung injury: role of oxidative stress. Microb. Pathog. 32, 27–34 (2002).

    Article  CAS  PubMed  Google Scholar 

  230. Auer, M., Pfister, L. A., Leppert, D., Tauber, M. G. & Leib, S. L. Effects of clinically used antioxidants in experimental pneumococcal meningitis. J. Infect. Dis. 182, 347–350 (2000).

    Article  CAS  PubMed  Google Scholar 

  231. de Gans, J. & van de Beek, D. Dexamethasone in adults with bacterial meningitis. N. Engl. J. Med. 347, 1549–1556 (2002).

    Article  CAS  PubMed  Google Scholar 

  232. Umeki, S. & Soejima, R. Hydrocortisone inhibits the respiratory burst oxidase from human neutrophils in whole-cell and cell-free systems. Biochim Biophys. Acta 1052, 211–215 (1990).

    Article  CAS  PubMed  Google Scholar 

  233. Korhonen, R., Lahti, A., Hamalainen, M., Kankaanranta, H. & Moilanen, E. Dexamethasone inhibits inducible nitric-oxide synthase expression and nitric oxide production by destabilizing mRNA in lipopolysaccharide-treated macrophages. Mol. Pharmacol. 62, 698–704 (2002).

    Article  CAS  PubMed  Google Scholar 

  234. Gao, X. P. et al. Role of NADPH oxidase in the mechanism of lung neutrophil sequestration and microvessel injury induced by Gram-negative sepsis: studies in p47phox−/− and gp91phox−/− mice. J. Immunol. 168, 3974–3982 (2002).

    Article  CAS  PubMed  Google Scholar 

  235. Morgenstern, D. E., Gifford, M. A., Li, L. L., Doerschuk, C. M. & Dinauer, M. C. Absence of respiratory burst in X-linked chronic granulomatous disease mice leads to abnormalities in both host defense and inflammatory response to Aspergillus fumigatus. J. Exp. Med. 185, 207–218 (1997). Shows that NADPH oxidase might be important in the resolution of inflammatory lesions.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  236. Weiss, J., Kao, L., Victor, M. & Elsbach, P. Respiratory burst facilitates the digestion of Escherichia coli killed by polymorphonuclear leukocytes. Infect. Immun. 55, 2142–2147 (1987).

    CAS  PubMed  PubMed Central  Google Scholar 

  237. Carson, M. J., Chadwick, D. L., Brubaker, C. A., Cleland, R. S. & Landing, B. H. Thirteen boys with progressive septic granulomatosis. Pediatrics 35, 405–412 (1965).

    CAS  PubMed  Google Scholar 

  238. Pignatelli, B. et al. Helicobacter pylori eradication attenuates oxidative stress in human gastric mucosa. Am. J. Gastroenterol. 96, 1758–1766 (2001).

    Article  CAS  PubMed  Google Scholar 

  239. Blanchard, T. G., Yu, F., Hsieh, C. L. & Redline, R. W. Severe inflammation and reduced bacteria load in murine helicobacter infection caused by lack of phagocyte oxidase activity. J. Infect. Dis. 187, 1609–1615 (2003).

    Article  CAS  PubMed  Google Scholar 

  240. Laroux, F. S. et al. Role of nitric oxide in the regulation of acute and chronic inflammation. Antioxid. Redox Signal. 2, 391–396 (2000).

    Article  CAS  PubMed  Google Scholar 

  241. Anstey, N. M. et al. Nitric oxide in Tanzanian children with malaria: inverse relationship between malaria severity and nitric oxide production/nitric oxide synthase type 2 expression. J. Exp. Med. 184, 557–567 (1996).

    Article  CAS  PubMed  Google Scholar 

  242. Matsushita, K. et al. Nitric oxide regulates exocytosis by S-nitrosylation of N-ethylmaleimide-sensitive factor. Cell 115, 139–150 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The author is deeply grateful for the insight and suggestions provided by M. Dinauer, S. Holland, C. Lowenstein, C. Nathan, W. Nauseef, H. Rosen and A. Vazquez-Torres, as well as the members of the Fang laboratory. Space limitations prevented the inclusion of many noteworthy citations — in such instances, the author has cited a representative article or referred the reader to a relevant review. Research support was provided by the US National Institutes of Health.

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  1. Departments of Laboratory Medicine and Microbiology, University of Washington School of Medicine, 1959 North East Pacific Street, Box 357242, Seattle, Washington, 98195-7242, USA

    Ferric C. Fang

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DATABASES

Infectious Disease Information

leishmaniasis

tuberculosis

OMIM

CGD

xanthinuria

SwissProt

DtxR

Fur

glucose-6-phosphate dehydrogenase

gp91-phox

iNOS

MPO

p22-phox

p47-phox

p67-phox

Rac1

Rac2

Rap1A

xanthine oxidase

YopE

YopH

FURTHER INFORMATION

Ferric C. Fang's laboratory

Glossary

PATHOPHYSIOLOGICAL

Functional changes that are associated with, or result from, disease or injury.

POLYMORPHONUCLEAR PHAGOCYTES

White blood cells with multi-lobed nuclei and cytoplasmic granules that are involved in inflammatory responses.

PROINFLAMMATORY CYTOKINES

Secreted proteins with autocrine or paracrine action that regulate the inflammatory response. There are many types of cytokine, which elicit different cellular responses, including the control of cell proliferation and differentiation, the regulation of immune responses and haematopoiesis.

AGONIST PEPTIDES

Peptides that mimic cognate antigen, which results in cellular activation.

ELECTROGENIC

Generating an electrical potential across a membrane.

DENDRITIC CELLS

'Professional' antigen-presenting cells that are found in the T-cell areas of lymphoid tissues and as minor cellular components in most tissues. They have a branched or dendritic morphology and are the most potent stimulators of T-cell responses.

ISOFORMS

Forms of a protein with slightly different amino-acid sequences that often have diverse activities, functions and/or distributions.

HYPOHALOUS

A compound in which a hydroxyl group is combined with a halogen atom.

FENTON REACTION

The reduction of hydrogen peroxide by ferrous iron.

PEROXIDATION

A type of reaction in which oxygen atoms are formed, which leads to the production of peroxides.

MYELOPEROXIDASE

Peroxidase from neutrophils that takes part in the bactericidal activity of these cells. The name originates from the first isolation from the blood of patients with myeloid leukaemia.

PROTEASOME

In eukaryotes the 26S proteasome is a large multisubunit protease complex that selectively degrades multi-ubiquitylated proteins. It contains a 20S particle that carries the catalytic activity and two regulatory 19S particles.

IONOPHORE

Small hydrophobic molecules that dissolve in lipid bilayers and increase the permeability of membranes to ions.

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Fang, F. Antimicrobial reactive oxygen and nitrogen species: concepts and controversies. Nat Rev Microbiol 2, 820–832 (2004). https://doi.org/10.1038/nrmicro1004

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