Assays for Inducing and Measuring Cell Death to Detect Macrophage Migration Inhibitory Factor (MIF) Release

  • Shahrzad Zamani
  • Eric F. MorandEmail author
  • Jacqueline K. Flynn
Part of the Methods in Molecular Biology book series (MIMB, volume 2080)


Cell death is a vital process for maintaining tissue homeostasis and removing potentially harmful cells. Cell death can be both programmed and non-programmed and is commonly divided into two main forms, termed apoptotic and necrotic death modes. In this chapter cell death is classified into apoptosis, primary necrosis, pyroptosis, and necroptosis. This chapter outlines the measurement of these different types of cell death and the relationship of measuring MIF release in these assays.

Key words

MIF Apoptosis Necrosis Pyroptosis Necroptosis Flow cytometry Colorimetric assay 


  1. 1.
    Calandra T, Roger T (2003) Macrophage migration inhibitory factor: a regulator of innate immunity. Nat Rev Immunol 3:791CrossRefGoogle Scholar
  2. 2.
    Bloom J, Sun S, Al-Abed Y (2016) MIF, a controversial cytokine: a review of structural features, challenges, and opportunities for drug development. Expert Opin Ther Targets 20(12):1463–1475CrossRefGoogle Scholar
  3. 3.
    Calandra T et al (1994) The macrophage is an important and previously unrecognized source of macrophage migration inhibitory factor. J Exp Med 179(6):1895–1902CrossRefGoogle Scholar
  4. 4.
    Rice EK et al (2003) Induction of MIF synthesis and secretion by tubular epithelial cells: a novel action of angiotensin II. Kidney Int 63(4):1265–1275CrossRefGoogle Scholar
  5. 5.
    Calandra T et al (2000) Protection from septic shock by neutralization of macrophage migration inhibitory factor. Nat Med 6:164CrossRefGoogle Scholar
  6. 6.
    Radstake TRDJ et al (2005) Correlation of rheumatoid arthritis severity with the genetic functional variants and circulating levels of macrophage migration inhibitory factor. Arthritis Rheum 52(10):3020–3029CrossRefGoogle Scholar
  7. 7.
    Kim HR et al (2007) Macrophage migration inhibitory factor upregulates angiogenic factors and correlates with clinical measures in rheumatoid arthritis. J Rheumatol 34(5):927–936PubMedGoogle Scholar
  8. 8.
    Foote A et al (2004) Macrophage migration inhibitory factor in systemic lupus erythematosus. J Rheumatol 31(2):268–273PubMedGoogle Scholar
  9. 9.
    Sánchez E et al (2006) Evidence of association of macrophage migration inhibitory factor gene polymorphisms with systemic lupus erythematosus. Genes Immun 7:433CrossRefGoogle Scholar
  10. 10.
    Kok T et al (2018) Small-molecule inhibitors of macrophage migration inhibitory factor (MIF) as an emerging class of therapeutics for immune disorders. Drug Discov Today 23(11):1910–1918CrossRefGoogle Scholar
  11. 11.
    Hertelendy J et al (2018) Macrophage migration inhibitory factor - a favorable marker in inflammatory diseases? Curr Med Chem 25(5):601–605CrossRefGoogle Scholar
  12. 12.
    Roth S et al (2015) Secondary necrotic neutrophils release interleukin-16C and macrophage migration inhibitory factor from stores in the cytosol. Cell Death Discov 1:15056–15056CrossRefGoogle Scholar
  13. 13.
    Roth S, Solbach W, Laskay T (2016) IL-16 and MIF: messengers beyond neutrophil cell death. Cell Death Dis 7(1):e2049–e2049CrossRefGoogle Scholar
  14. 14.
    Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35(4):495–516CrossRefGoogle Scholar
  15. 15.
    Jorgensen I, Rayamajhi M, Miao EA (2017) Programmed cell death as a defence against infection. Nat Rev Immunol 17:151CrossRefGoogle Scholar
  16. 16.
    Cohen JJ (1991) Programmed cell death in the immune system. Adv Immunol 50:55–85CrossRefGoogle Scholar
  17. 17.
    Yang Y et al (2015) Programmed cell death and its role in inflammation. Mil Med Res 2(1):12CrossRefGoogle Scholar
  18. 18.
    Choi JJ, Reich Iii CF, Pisetsky DS (2004) Release of DNA from dead and dying lymphocyte and monocyte cell lines in vitro. Scand J Immunol 60(1–2):159–166CrossRefGoogle Scholar
  19. 19.
    Vermes I et al (1995) A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 184(1):39–51CrossRefGoogle Scholar
  20. 20.
    Yurinskaya V et al (2017) A comparative study of U937 cell size changes during apoptosis initiation by flow cytometry, light scattering, water assay and electronic sizing. Apoptosis 22(10):1287–1295CrossRefGoogle Scholar
  21. 21.
    Castilla R et al (2004) Dual effect of ethanol on cell death in primary culture of human and rat hepatocytes. Alcohol Alcohol 39(4):290–296CrossRefGoogle Scholar
  22. 22.
    Tait SWG, Ichim G, Green DR (2014) Die another way--non-apoptotic mechanisms of cell death. J Cell Sci 127(Pt 10):2135–2144CrossRefGoogle Scholar
  23. 23.
    Bergsbaken T, Fink SL, Cookson BT (2009) Pyroptosis: host cell death and inflammation. Nat Rev Microbiol 7(2):99–109CrossRefGoogle Scholar
  24. 24.
    Man SM, Karki R, Kanneganti TD (2017) Molecular mechanisms and functions of pyroptosis, inflammatory caspases and inflammasomes in infectious diseases. Immunol Rev 277(1):61–75CrossRefGoogle Scholar
  25. 25.
    Magna M, Pisetsky D (2015) The role of cell death in the pathogenesis of SLE: Is pyroptosis the missing link? Scand J Immunol 82(3):218–224CrossRefGoogle Scholar
  26. 26.
    Aglietti RA, Dueber EC (2017) Recent insights into the molecular mechanisms underlying pyroptosis and gasdermin family functions. Trends Immunol 38(4):261–271CrossRefGoogle Scholar
  27. 27.
    Vanaja SK, Rathinam VA, Fitzgerald KA (2015) Mechanisms of inflammasome activation: recent advances and novel insights. Trends Cell Biol 25(5):308–315CrossRefGoogle Scholar
  28. 28.
    He Y, Hara H, Nunez G (2016) Mechanism and regulation of NLRP3 inflammasome activation. Trends Biochem Sci 41(12):1012–1021CrossRefGoogle Scholar
  29. 29.
    Lang T et al (2018) Macrophage migration inhibitory factor is required for NLRP3 inflammasome activation. Nat Commun 9(1):2223CrossRefGoogle Scholar
  30. 30.
    Dhuriya YK, Sharma D (2018) Necroptosis: a regulated inflammatory mode of cell death. J Neuroinflammation 15(1):199CrossRefGoogle Scholar
  31. 31.
    Omoto S et al (2015) Suppression of RIP3-dependent necroptosis by human cytomegalovirus. J Biol Chem 290(18):11635–11648CrossRefGoogle Scholar
  32. 32.
    Murphy JM, Silke J (2014) Ars Moriendi; the art of dying well–new insights into the molecular pathways of necroptotic cell death. EMBO Rep 15(2):155–164CrossRefGoogle Scholar
  33. 33.
    Hanson B (2016) Necroptosis: a new way of dying? Cancer Biol Ther 17(9):899–910CrossRefGoogle Scholar
  34. 34.
    Kearney CJ, Martin SJ (2017) An inflammatory perspective on necroptosis. Mol Cell 65(6):965–973CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Shahrzad Zamani
    • 1
  • Eric F. Morand
    • 2
    Email author
  • Jacqueline K. Flynn
    • 2
  1. 1.Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical ResearchMonash UniversityClaytonAustralia
  2. 2.Rheumatology Group, Centre for Inflammatory Diseases, Department of Medicine, School of Clinical Sciences at Monash Health, Faculty of Medicine, Nursing & Health SciencesMonash University, Monash Medical CentreClaytonAustralia

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