Advertisement

Diagnostic Assays for Myeloperoxidase and Myeloperoxidase Deficiency

  • William M. Nauseef
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1124)

Abstract

Neutrophils (PMN) represent the dominant cell in the acute response to microbial infection and can contribute to some of the tissue damage that accompanies sterile inflammation. Effective antimicrobial activity in neutrophil phagosomes reflects the combined action of soluble agents in plasma with PMN-derived reactive oxygen species and granule proteins, including the azurophilic granule protein myeloperoxidase (MPO). The inhibition or the absence of the MPO–H2O2–halide system results in marked reduction in PMN killing of a variety of microbes, implicating its relative prominence in the hierarchy of PMN antimicrobial systems. Although the most profound clinical defects are manifested in patients lacking the capacity to generate reactive oxygen species, as seen in chronic granulomatous disease, an inherited deficiency of MPO can also increase the frequency or the severity of clinical infections.

Like related peroxidases expressed in animals, MPO can catalyze both one- and two-electron oxidations, thereby mediating peroxidation and halogenation, respectively. The presence of each activity can be assessed in inflammatory fluids or by stimulated PMN. Furthermore, histochemical staining provides an assessment of functional MPO in tissue or within PMN, and immunoblotting of isolated PMN for MPO can provide additional insight into the molecular basis of the observed absence of functional enzyme.

Keywords

Peroxidase Myeloperoxidase Eosinophil peroxidase Azurophilic granules 

References

  1. 1.
    Klebanoff SJ, Kettle AJ, Rosen H et al (2013) Myeloperoxidase: a front-line defender against phagocytosed microorganisms. J Leukoc Biol 93:185–198PubMedCrossRefGoogle Scholar
  2. 2.
    Hurst JK (2012) What really happens in the neutrophil phagosome? Free Radic Biol Med 53:508–520PubMedCrossRefGoogle Scholar
  3. 3.
    Dinauer MC, Nauseef WM, Newburger PE (2001) Inherited disorders of phagocyte killing. In: Scriver CR, Beaudet al, Valle D et al (eds) The metabolic and molecular bases of inherited diseases, vol 8. McGraw-Hill Companies, New York, pp 4857–4887Google Scholar
  4. 4.
    Mauch L, Lun A, O’Gorman MRG et al (2007) Chronic granulomatous disease (CGD) and complete myeloperoxidase deficiency both yield strongly reduced dihydrorhodamine 123 test signals but can be easily discerned in routine testing for CGD. Clin Chem 53:890–896PubMedCrossRefGoogle Scholar
  5. 5.
    Kettle AJ, Winterbourn CC (1994) Assays for the chlorination activity of myeloperoxidase. Methods Enzymol 233:502–512PubMedCrossRefGoogle Scholar
  6. 6.
    Kettle AJ (1999) Detection of 3-chlorotyrosine in proteins exposed to neutrophil oxidants. Methods Enzymol 300:111–120PubMedCrossRefGoogle Scholar
  7. 7.
    Dypbukt JM, Bishop C, Brooks WM et al (2005) A sensitive and selective assay for chloramine production by myeloperoxidase. Free Radic Biol Med 39:1468–1477PubMedCrossRefGoogle Scholar
  8. 8.
    Mage MM, Evans WH, Himmelhoch SR et al (1971) Immunological identification and quantification of guinea pig heterophil and eosinophil peroxidases. J Reticuloendothel Soc 9:201–208PubMedGoogle Scholar
  9. 9.
    Fabian I, Aronson M (1975) Deamination of histamine by peroxidase of neutrophils and eosinophils. J Reticuloendothel Soc 17: 141–145PubMedGoogle Scholar
  10. 10.
    Carlson MG, Peterson CG, Venge P (1985) Human eosinophil peroxidase: purification and characterization. J Immunol 134:1875–1879PubMedGoogle Scholar
  11. 11.
    Dri P, Cramer R, Soranzo MR et al (1982) New approaches to the detection of myeloperoxidase deficiency. Blood 60:323–327PubMedGoogle Scholar
  12. 12.
    Hansson M, Olsson I, Nauseef WM (2006) Biosynthesis, processing, and sorting of human myeloperoxidase. Arch Biochem Biophys 445: 214–224PubMedCrossRefGoogle Scholar
  13. 13.
    Taylor KL, Pohl J, Kinkade JM Jr (1992) Unique autolytic cleavage of human myeloperoxidase. J Biol Chem 267:25282–25288PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • William M. Nauseef
    • 1
  1. 1.Inflammation Program and Department of MedicineRoy J.and Lucille A. Carver College of Medicine, University of IowaCoralvilleUSA

Personalised recommendations