Host control of infections crucially depends on the capability to kill pathogens with reactive oxygen species (ROS). However, these toxic molecules can also readily damage host components and cause severe immunopathology. Here, we show that neutrophils use their most abundant granule protein, myeloperoxidase, to target ROS specifically to pathogens while minimizing collateral tissue damage. A computational model predicted that myeloperoxidase efficiently scavenges diffusible H2O2 at the surface of phagosomal Salmonella and converts it into highly reactive HOCl (bleach), which rapidly damages biomolecules within a radius of less than 0.1 μm. Myeloperoxidase-deficient neutrophils were predicted to accumulate large quantities of H2O2 that still effectively kill Salmonella, but most H2O2 would leak from the phagosome. Salmonella stimulation of neutrophils from normal and myeloperoxidase-deficient human donors experimentally confirmed an inverse relationship between myeloperoxidase activity and extracellular H2O2 release. Myeloperoxidase-deficient mice infected with Salmonella had elevated hydrogen peroxide tissue levels and exacerbated oxidative damage of host lipids and DNA, despite almost normal Salmonella control. These data show that myeloperoxidase has a major function in mitigating collateral tissue damage during antimicrobial oxidative bursts, by converting diffusible long-lived H2O2 into highly reactive, microbicidal and locally confined HOCl at pathogen surfaces.
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Klebanoff, S. J., Kettle, A. J., Rosen, H., Winterbourn, C. C. & Nauseef, W. M. Myeloperoxidase: a front-line defender against phagocytosed microorganisms. J. Leukoc. Biol. 93, 185–198 (2013).
Arnhold, J., Furtmüller, P. G., Regelsberger, G. & Obinger, C. Redox properties of the couple compound I/native enzyme of myeloperoxidase and eosinophil peroxidase. Eur. J. Biochem. 268, 5142–5148 (2001).
Storkey, C., Davies, M. J. & Pattison, D. I. Reevaluation of the rate constants for the reaction of hypochlorous acid (HOCl) with cysteine, methionine, and peptide derivatives using a new competition kinetic approach. Free Radic. Biol. Med. 73, 60–66 (2014).
Parker, H. & Winterbourn, C. C. Reactive oxidants and myeloperoxidase and their involvement in neutrophil extracellular traps. Front. Immunol. 3, 424 (2013).
Metzler, K. D., Goosmann, C., Lubojemska, A., Zychlinsky, A. & Papayannopoulos, V. A myeloperoxidase-containing complex regulates neutrophil elastase release and actin dynamics during NETosis. Cell Rep. 8, 883–896 (2014).
Davies, M. J., Hawkins, C. L., Pattison, D. I. & Rees, M. D. Mammalian heme peroxidases: from molecular mechanisms to health implications. Antioxid. Redox Signal. 10, 1199–1234 (2008).
Ruggeri, R. B. et al. Discovery of 2-(6-(5-chloro-2-methoxyphenyl)-4-oxo-2-thioxo-3,4-dihydropyrimidin-1(2H)-yl)acet amide (PF-06282999): a highly selective mechanism-based myeloperoxidase inhibitor for the treatment of cardiovascular diseases. J. Med. Chem. 58, 8513–8528 (2015).
Winterbourn, C. C., Hampton, M. B., Livesey, J. H. & Kettle, A. J. Modeling the reactions of superoxide and myeloperoxidase in the neutrophil phagosome: implications for microbial killing. J. Biol. Chem. 281, 39860–39869 (2006).
Burton, N. A. et al. Disparate impact of oxidative host defenses determines the fate of Salmonella during systemic infection in mice. Cell Host Microbe 15, 72–83 (2014).
Imlay, J. A. in EcoSal (eds Curtiss, R. et al.) Module 5.4.4 (ASM, 2009).
Seaver, L. C. & Imlay, J. A. Hydrogen peroxide fluxes and compartmentalization inside growing Escherichia coli. J. Bacteriol. 183, 7182–7189 (2001).
Makino, N., Sasaki, K., Hashida, K. & Sakakura, Y. A metabolic model describing the H2O2 elimination by mammalian cells including H2O2 permeation through cytoplasmic and peroxisomal membranes: comparison with experimental data. Biochim. Biophys. Acta 1673, 149–159 (2004).
Kundrat, P., Bauer, G., Jacob, P. & Friedland, W. Mechanistic modelling suggests that the size of preneoplastic lesions is limited by intercellular induction of apoptosis in oncogenically transformed cells. Carcinogenesis 33, 253–259 (2012).
Green, J. N., Kettle, A. J. & Winterbourn, C. C. Protein chlorination in neutrophil phagosomes and correlation with bacterial killing. Free Radic. Biol. Med. 77, 49–56 (2014).
Korshunov, S. S. & Imlay, J. A. A potential role for periplasmic superoxide dismutase in blocking the penetration of external superoxide into the cytosol of Gram-negative bacteria. Mol. Microbiol. 43, 95–106 (2002).
Stroppolo, M. E. et al. Single mutation at the intersubunit interface confers extra efficiency to Cu,Zn superoxide dismutase. FEBS Lett. 483, 17–20 (2000).
Bull, C. & Fee, J. A. Steady-state kinetic studies of superoxide dismutases: properties of the iron containing protein from Escherichia coli. J. Am. Chem. Soc. 107, 3295–3304 (1985).
Park, S., You, X. & Imlay, J. A. Substantial DNA damage from submicromolar intracellular hydrogen peroxide detected in Hpx- mutants of Escherichia coli. Proc. Natl Acad. Sci. USA 102, 9317–9322 (2005).
Winterbourn, C. C. Reconciling the chemistry and biology of reactive oxygen species. Nat. Chem. Biol. 4, 278–286 (2008).
Benfeitas, R., Selvaggio, G., Antunes, F., Coelho, P. M. & Salvador, A. Hydrogen peroxide metabolism and sensing in human erythrocytes: a validated kinetic model and reappraisal of the role of peroxiredoxin II. Free Radic. Biol. Med. 74, 35–49 (2014).
Nauseef, W. M., Metcalf, J. A. & Root, R. K. Role of myeloperoxidase in the respiratory burst of human neutrophils. Blood 61, 483–492 (1983).
Gerber, C. E., Kuci, S., Zipfel, M., Niethammer, D. & Bruchelt, G. Phagocytic activity and oxidative burst of granulocytes in persons with myeloperoxidase deficiency. Eur. J. Clin. Chem. Clin. Biochem. 34, 901–908 (1996).
Gross, S. et al. Bioluminescence imaging of myeloperoxidase activity in vivo. Nat. Med. 15, 455–461 (2009).
Kettle, A. J., Gedye, C. A., Hampton, M. B. & Winterbourn, C. C. Inhibition of myeloperoxidase by benzoic acid hydrazides. Biochem. J. 308(Pt 2), 559–563 (1995).
Flemmig, J., Remmler, J., Zschaler, J. & Arnhold, J. Detection of the halogenating activity of heme peroxidases in leukocytes by aminophenyl fluorescein. Free Radic. Res. 49, 768–776 (2015).
Naegelen, I. et al. An essential role of syntaxin 3 protein for granule exocytosis and secretion of IL-1α, IL-1β, IL-12β, and CCL4 from differentiated HL-60 cells. J. Leukoc. Biol. 97, 557–571 (2015).
Winterbourn, C. C., Kettle, A. J. & Hampton, M. B. Reactive oxygen species and neutrophil function. Annu. Rev. Biochem. 85, 765–792 (2016).
Tsolis, R. M., Xavier, M. N., Santos, R. L. & Baumler, A. J. How to become a top model: the impact of animal experimentation on human Salmonella disease research. Infect. Immun. 79, 1806–1814 (2011).
Conlan, J. W. Critical roles of neutrophils in host defense against experimental systemic infections of mice by Listeria monocytogenes, Salmonella typhimurium, and Yersinia enterocolitica. Infect. Immun. 65, 630–635 (1997).
Vassiloyanakopoulos, A. P., Okamoto, S. & Fierer, J. The crucial role of polymorphonuclear leukocytes in resistance to Salmonella Dublin infections in genetically susceptible and resistant mice. Proc. Natl Acad. Sci. USA 95, 7676–7681 (1998).
Cheminay, C., Chakravortty, D. & Hensel, M. Role of neutrophils in murine salmonellosis. Infect. Immun. 72, 468–477 (2004).
Dejager, L., Pinheiro, I., Bogaert, P., Huys, L. & Libert, C. Role for neutrophils in host immune responses and genetic factors that modulate resistance to Salmonella enterica serovar typhimurium in the inbred mouse strain SPRET/Ei. Infect. Immun. 78, 3848–3860 (2010).
Mastroeni, P. et al. Resistance and susceptibility to Salmonella infections lessons from mice and patients with immunodeficiencies. Rev. Med. Microbiol. 14, 53–62 (2003).
Lee, C. et al. Redox regulation of OxyR requires specific disulfide bond formation involving a rapid kinetic reaction path. Nat. Struct. Mol. Biol. 11, 1179–1185 (2004).
Aslund, F., Zheng, M., Beckwith, J. & Storz, G. Regulation of the OxyR transcription factor by hydrogen peroxide and the cellular thiol-disulfide status. Proc. Natl Acad. Sci. USA 96, 6161–6165 (1999).
Swirski, F. K. et al. Myeloperoxidase-rich Ly-6C+ myeloid cells infiltrate allografts and contribute to an imaging signature of organ rejection in mice. J. Clin. Invest. 120, 2627–2634 (2010).
Kremserova, S. et al. Lung neutrophilia in myeloperoxidase deficient mice during the course of acute pulmonary inflammation. Oxid. Med. Cell Longev. 2016, 5219056 (2016).
Endo, D., Saito, T., Umeki, Y., Suzuki, K. & Aratani, Y. Myeloperoxidase negatively regulates the expression of proinflammatory cytokines and chemokines by zymosan-induced mouse neutrophils. Inflamm. Res. 65, 151–159 (2016).
Brovkovych, V. et al. Augmented inducible nitric oxide synthase expression and increased NO production reduce sepsis-induced lung injury and mortality in myeloperoxidase-null mice. Am. J. Physiol. Lung Cell Mol. Physiol. 295, L96–L103 (2008).
Homme, M., Tateno, N., Miura, N., Ohno, N. & Aratani, Y. Myeloperoxidase deficiency in mice exacerbates lung inflammation induced by nonviable Candida albicans. Inflamm. Res. 62, 981–990 (2013).
Sugamata, R. et al. Contribution of neutrophil-derived myeloperoxidase in the early phase of fulminant acute respiratory distress syndrome induced by influenza virus infection. Microbiol. Immunol. 56, 171–182 (2012).
Takeuchi, K. et al. Severe neutrophil-mediated lung inflammation in myeloperoxidase-deficient mice exposed to zymosan. Inflamm. Res. 61, 197–205 (2012).
Brennan, M. L. et al. A tale of two controversies: defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase-deficient mice, and the nature of peroxidase-generated reactive nitrogen species. J. Biol. Chem. 277, 17415–17427 (2002).
Klinke, A. et al. Myeloperoxidase attracts neutrophils by physical forces. Blood 117, 1350–1358 (2011).
Imlay, J. A. The molecular mechanisms and physiological consequences of oxidative stress: lessons from a model bacterium. Nat. Rev. Microbiol. 11, 443–454 (2013).
Liou, G. Y. & Storz, P. Detecting reactive oxygen species by immunohistochemistry. Methods Mol. Biol. 1292, 97–104 (2015).
Seki, S. et al. In situ detection of lipid peroxidation and oxidative DNA damage in non-alcoholic fatty liver diseases. J. Hepatol. 37, 56–62 (2002).
Loft, S. & Poulsen, H. E. Cancer risk and oxidative DNA damage in man. J. Mol. Med. (Berl.) 74, 297–312 (1996).
Finkel, T. & Holbrook, N. J. Oxidants, oxidative stress and the biology of ageing. Nature 408, 239–247 (2000).
Lanza, F. Clinical manifestation of myeloperoxidase deficiency. J. Mol. Med. (Berl.) 76, 676–681 (1998).
Yuzhalin, A. E. & Kutikhin, A. G. Common genetic variants in the myeloperoxidase and paraoxonase genes and the related cancer risk: a review. J. Environ. Sci. Health C 30, 287–322 (2012).
Crimmins, E. M. & Finch, C. E. Infection, inflammation, height, and longevity. Proc. Natl Acad. Sci. USA 103, 498–503 (2006).
Zhang, R. et al. Myeloperoxidase functions as a major enzymatic catalyst for initiation of lipid peroxidation at sites of inflammation. J. Biol. Chem. 277, 46116–46122 (2002).
Kolaczkowska, E., Koziol, A., Plytycz, B. & Arnold, B. Inflammatory macrophages, and not only neutrophils, die by apoptosis during acute peritonitis. Immunobiology 215, 492–504 (2010).
Grant, A. J. et al. Caspase-3-dependent phagocyte death during systemic Salmonella enterica serovar Typhimurium infection of mice. Immunology 125, 28–37 (2008).
Branzk, N. et al. Neutrophils sense microbe size and selectively release neutrophil extracellular traps in response to large pathogens. Nat. Immunol. 15, 1017–1025 (2014).
Kutter, D. et al. Consequences of total and subtotal myeloperoxidase deficiency: risk or benefit? Acta Haematol. 104, 10–15 (2000).
Hjorth, R., Jonsson, A. K. & Vretblad, P. A rapid method for purification of human granulocytes using percoll. A comparison with dextran sedimentation. J. Immunol. Methods 43, 95–101 (1981).
Hoiseth, S. K. & Stocker, B. A. Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature 291, 238–239 (1981).
Kroger, C. et al. The transcriptional landscape and small RNAs of Salmonella enterica serovar Typhimurium. Proc. Natl Acad. Sci. USA 109, E1277–E1286 (2012).
de Chaumont, F. et al. Icy: an open bioimage informatics platform for extended reproducible research. Nat. Methods 9, 690–696 (2012).
Huang, L.-K. & Wang, M.-J. J. Image thresholding by minimizing the measures of fuzziness. Pattern Recognit. 28, 41–51 (1995).
The authors thank K. Ullrich and R. Kühl for taking blood from human donors, and thank all donors for blood donations. The authors thank I. Bartholomaeus and A. Martin for support with confocal microscopy. This study was supported in part by grants from the Swiss National Foundation (310030_156818 to D.B., PZ00P3_142403 to N.K. and PP00P3_144863 to M.R.) and the Gebert Rüf Foundation (GRS 058/14 to C.H., A.-V.B. and M.R.).
The authors declare no competing financial interests.
About this article
Cite this article
Schürmann, N., Forrer, P., Casse, O. et al. Myeloperoxidase targets oxidative host attacks to Salmonella and prevents collateral tissue damage. Nat Microbiol 2, 16268 (2017). https://doi.org/10.1038/nmicrobiol.2016.268
Distinct macrophage phenotypes and redox environment during the fin fold regenerative process in zebrafish
Scandinavian Journal of Immunology (2021)
Myeloperoxidase: A versatile mediator of endothelial dysfunction and therapeutic target during cardiovascular disease
Pharmacology & Therapeutics (2021)
The effects of neutrophil-generated hypochlorous acid and other hypohalous acids on host and pathogens
Cellular and Molecular Life Sciences (2020)
Escaping the Phagocytic Oxidative Burst: The Role of SODB in the Survival of Pseudomonas aeruginosa Within Macrophages
Frontiers in Microbiology (2020)