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Evaluation of Classical, Alternative, and Regulatory Functions of Bone Marrow-Derived Macrophages

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Mouse Models of Allergic Disease

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1032))

Abstract

The role of macrophage subsets in allergic diseases in vivo is under current investigation. These cells perform sentinel functions in the lung, the skin, and the gastrointestinal mucosa. Their interface with environmental cues influences the initiation, progression, development, and resolution of allergic diseases. Researchers often culture bone marrow-derived macrophages to study macrophage biology. The in vitro maturation of bone marrow precursor cells into mature macrophages is a powerful technique used to study macrophage biology. The polarization or differential activation of macrophages into functionally distinct subsets can provide insight into allergic disease pathologies. Classically activated, alternatively activated, and regulatory macrophages have different effector functions that can affect allergic responses. Understanding macrophage biology during allergen exposure, host sensitization, and disease progression/resolution may lead to improved therapeutic interventions. The purpose of this chapter is to outline protocols used for the culture and polarization of classically activated, alternatively activated, and regulatory macrophages. In addition, the techniques to measure macrophage-specific effector molecules by ELISA, RT-PCR, and immunoblotting are reviewed.

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References

  1. Murray PJ, Wynn TA (2011) Protective and pathogenic functions of macrophage subsets. Nat Rev Immunol 11(11):723–737

    Article  PubMed  CAS  Google Scholar 

  2. Moreira AP, Hogaboam CM (2011) Macrophages in allergic asthma: fine-tuning their pro- and anti-inflammatory actions for disease resolution. J Interferon Cytokine Res 31(6):485–491

    Article  PubMed  CAS  Google Scholar 

  3. Palm NW, Rosenstein RK, Medzhitov R (2012) Allergic host defenses. Nature 484:465–472

    Article  PubMed  CAS  Google Scholar 

  4. Holgate ST (2012) Innate and adaptive immune responses in asthma. Nat Med 18:673–683

    Article  PubMed  CAS  Google Scholar 

  5. Holgate ST (2011) Pathophysiology of asthma: what has our current understanding taught us about new therapeutic approaches? J Allergy Clin Immunol 128(3):495–505

    Article  PubMed  CAS  Google Scholar 

  6. Gordon S (2007) The macrophage: past, present and future. Eur J Immunol 37 Suppl: S9–S17

    Google Scholar 

  7. Gordon S, Martinez FO (2010) Alternative activation of macrophages: mechanism and functions. Immunity 32(5):593–604

    Article  PubMed  CAS  Google Scholar 

  8. Mantovani A (2006) Macrophage diversity and polarization: in vivo veritas. Blood 108(2):408–409

    Article  CAS  Google Scholar 

  9. Sica A, Mantovani A (2012) Macrophage plasticity and polarization: in vivo veritas. J Clin Invest 122(3):787–795

    Article  PubMed  CAS  Google Scholar 

  10. Lawrence T, Natoli G (2011) Transcriptional regulation of macrophage polarization: enabling diversity with identity. Nat Rev Immunol 11(11):750–761

    Article  PubMed  CAS  Google Scholar 

  11. Moreira AP, Cavassani KA, Hullinger R et al (2010) Serum amyloid P attenuates M2 macrophage activation and protects against fungal spore-induced allergic airway disease. J Allergy Clin Immunol 126(4):712–721.e7

    Article  PubMed  CAS  Google Scholar 

  12. Bedoret D, Wallemacq H, Marichal T et al (2009) Lung interstitial macrophages alter dendritic cell functions to prevent airway allergy in mice. J Clin Invest 119(12):3723–3738

    Article  PubMed  CAS  Google Scholar 

  13. Shahid SK, Kharitonov SA, Wilson NM et al (2002) Increased interleukin-4 and decreased interferon-γ in exhaled breath condensate of children with asthma. Am J Respir Crit Care Med 165(9):1290–1293

    Article  PubMed  Google Scholar 

  14. Kim CK, Kim SW, Park CS et al (2003) Bronchoalveolar lavage cytokine profiles in acute asthma and acute bronchiolitis. J Allergy Clin Immunol 112(1):64–71

    Article  PubMed  CAS  Google Scholar 

  15. Edwards JP, Zhang X, Frauwirth KA, Mosser DM (2006) Biochemical and functional characterization of three activated macrophage populations. J Leukoc Biol 80(6):1298–1307

    Article  PubMed  CAS  Google Scholar 

  16. Mosser DM, Zhang X (2008) Activation of murine macrophages. Curr Protoc Immunol Chapter 14: Unit 14.2

    Google Scholar 

  17. Mosser DM, Edwards JP (2008) Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8(12):958–969

    Article  PubMed  CAS  Google Scholar 

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Davis, B.K. (2013). Evaluation of Classical, Alternative, and Regulatory Functions of Bone Marrow-Derived Macrophages. In: Allen, I. (eds) Mouse Models of Allergic Disease. Methods in Molecular Biology, vol 1032. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-496-8_6

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  • DOI: https://doi.org/10.1007/978-1-62703-496-8_6

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-495-1

  • Online ISBN: 978-1-62703-496-8

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