Advertisement

Isolation of Group 2 Innate Lymphoid Cells from Mouse Lungs

  • Claudia U. Duerr
  • Jörg H. Fritz
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1656)

Abstract

The recently described group 2 innate lymphoid cells (ILC2) exert critical roles in type 2 immune responses, epithelial repair at mucosal tissues and metabolic homeostasis. ILC2 release large amounts of type 2 cytokines such as interleukin 4 (IL-4), IL-5, and IL-13, driving type 2 immunity such as the defense against helminths. However, if not tightly regulated ILC2 can trigger unwanted type 2 immunopathologies including allergic airway inflammation, airway hyper-responsiveness, and atopic dermatitis. Viral respiratory tract infections, archetypal triggers of type 1 immune responses, often give rise to pulmonary type 2 immunopathologies such as asthma and asthma exacerbations. Interestingly, pulmonary viral infections induce the release of IL-33, followed by induction of ILC2-mediated pulmonary type 2 immunopathology independent of the adaptive immune system. Due to their scarcity at steady state but also after infection and inflammation, pulmonary ILC2 are challenging to work with. In this chapter, we describe the detection and isolation procedure of pulmonary mouse ILC2 by flow cytometry and compare four distinct enzymatic mouse lung tissue processing protocols for optimized cell yield.

Key words

Group 2 innate lymphoid cells Lung Respiratory virus infections IL-33 Mucosal immunity Innate type 2 immune response 

References

  1. 1.
    Hansel TT, Johnston SL, Openshaw PJ (2013) Microbes and mucosal immune responses in asthma. Lancet 381:861–873CrossRefGoogle Scholar
  2. 2.
    Wynn TA (2015) Type 2 cytokines: mechanisms and therapeutic strategies. Nat Rev Immunol 15:271–282CrossRefGoogle Scholar
  3. 3.
    Edwards MR, Bartlett NW, Hussell T, Openshaw P, Johnston SL (2012) The microbiology of asthma. Nat Rev Microbiol 10:459–471CrossRefGoogle Scholar
  4. 4.
    Artis D, Spits H (2015) The biology of innate lymphoid cells. Nature 517:293–301CrossRefGoogle Scholar
  5. 5.
    Paul WE, Zhu J (2010) How are T(H)2-type immune responses initiated and amplified? Nat Rev Immunol 10:225–235CrossRefGoogle Scholar
  6. 6.
    Moro K et al (2010) Innate production of T(H)2 cytokines by adipose tissue-associated c-kit(+)Sca-1(+) lymphoid cells. Nature 463:540–544CrossRefGoogle Scholar
  7. 7.
    Neill DR et al (2010) Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity. Nature 464:1367–1370CrossRefGoogle Scholar
  8. 8.
    Price AE et al (2010) Systemically dispersed innate IL-13-expressing cells in type 2 immunity. Proc Natl Acad Sci U S A 107:11489–11494CrossRefGoogle Scholar
  9. 9.
    McKenzie AN, Spits H, Eberl G (2014) Innate lymphoid cells in inflammation and immunity. Immunity 41:366–374CrossRefGoogle Scholar
  10. 10.
    Diefenbach A, Colonna M, Koyasu S (2014) Development, differentiation, and diversity of innate lymphoid cells. Immunity 41:354–365CrossRefGoogle Scholar
  11. 11.
    Huang Y et al (2015) IL-25-responsive, lineage-negative KLRG1(hi) cells are multipotential 'inflammatory' type 2 innate lymphoid cells. Nat Immunol 16:161–169CrossRefGoogle Scholar
  12. 12.
    Koyasu S (2015) Inflammatory ILC2 cells: disguising themselves as progenitors? Nat Immunol 16:133–134CrossRefGoogle Scholar
  13. 13.
    Gause WC, Wynn TA, Allen JE (2013) Type 2 immunity and wound healing: evolutionary refinement of adaptive immunity by helminths. Nat Rev Immunol 13:607–614CrossRefGoogle Scholar
  14. 14.
    Nussbaum JC et al (2013) Type 2 innate lymphoid cells control eosinophil homeostasis. Nature 502:245–248CrossRefGoogle Scholar
  15. 15.
    Halim TY et al (2014) Group 2 innate lymphoid cells are critical for the initiation of adaptive T helper 2 cell-mediated allergic lung inflammation. Immunity 40:425–435CrossRefGoogle Scholar
  16. 16.
    Duerr CU, Fritz JH (2016) Regulation of group 2 innate lymphoid cells. Cytokine 87:1–8CrossRefGoogle Scholar
  17. 17.
    Chang YJ et al (2011) Innate lymphoid cells mediate influenza-induced airway hyper-reactivity independently of adaptive immunity. Nat Immunol 12:631–638CrossRefGoogle Scholar
  18. 18.
    Monticelli LA et al (2011) Innate lymphoid cells promote lung-tissue homeostasis after infection with influenza virus. Nat Immunol 12:1045–1054CrossRefGoogle Scholar
  19. 19.
    Duerr CU et al (2016) Type I interferon restricts type 2 immunopathology through the regulation of group 2 innate lymphoid cells. Nat Immunol 17:65–75CrossRefGoogle Scholar
  20. 20.
    Moro K et al (2016) Interferon and IL-27 antagonize the function of group 2 innate lymphoid cells and type 2 innate immune responses. Nat Immunol 17:76–86CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Microbiology and ImmunologyMcGill University Research Centre on Complex Traits (MRCCT), McGill UniversityMontrealCanada
  2. 2.Department of PhysiologyMcGill University Research Centre on Complex Traits (MRCCT), McGill UniversityMontrealCanada

Personalised recommendations