Advertisement

Protocol for Determining the Effect of Neuroendocrine Hormones on Murine ILC Function

  • Linda Quatrini
  • Sophie UgoliniEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2121)

Abstract

Neuroendocrine hormones are recognized as important mediators of inflammation that participate in the regulation of the magnitude and length of the immune response. It was demonstrated that endogenous glucocorticoids control the function of innate lymphoid cells (ILCs), and this regulatory mechanism is both cell type- and tissue-specific and is required for host protection during infections. We describe here how to analyze in vitro the effects of corticosterone on murine ILCs, using flow cytometry. The protocols described allow for the identification of the specific combination of stimuli with which glucocorticoids cooperate to regulate the function of ILCs. These methods are instrumental to understanding the molecular mechanisms downstream of glucocorticoid receptor activation and can explain the tissue specificity of ILC response to glucocorticoids.

Key words

NK cells ILCs Glucocorticoids Flow cytometry IFN-γ Cytotoxic granule exocytosis 

Notes

Acknowledgments

S. Ugolini lab is supported by institutional grants from INSERM, CNRS, Aix-Marseille University and Marseille-Immunopole to the CIML and received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program, under grant agreement No. 648768; from the Agence Nationnale de la Recherche (ANR) (No. ANR-14-CE14-0009-01) and from the ARC foundation (No. PGA120140200817). L. Quatrini has received funding from AIRC and from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 800924.

References

  1. 1.
    Weikum ER, Knuesel MT, Ortlund EA et al (2017) Glucocorticoid receptor control of transcription: precision and plasticity via allostery. Nat Rev Mol Cell Biol 18:159–174CrossRefGoogle Scholar
  2. 2.
    Cain DW, Cidlowski JA (2017) Immune regulation by glucocorticoids. Nat Rev Immunol 17:233–247CrossRefGoogle Scholar
  3. 3.
    Quatrini L, Wieduwild E, Guia S et al (2017) Host resistance to endotoxic shock requires the neuroendocrine regulation of group 1 innate lymphoid cells. J Exp Med 214:3531–3541CrossRefGoogle Scholar
  4. 4.
    Quatrini L, Wieduwild E, Escaliere B et al (2018) Endogenous glucocorticoids control host resistance to viral infection through the tissue-specific regulation of PD-1 expression on NK cells. Nat Immunol 19:954–962CrossRefGoogle Scholar
  5. 5.
    Vivier E, Tomasello E, Baratin M et al (2008) Functions of natural killer cells. Nat Immunol 9:503–510CrossRefGoogle Scholar
  6. 6.
    Shapiro HM (1993) Trends and developments in flow cytometry instrumentation. Ann N Y Acad Sci 677:155–166CrossRefGoogle Scholar
  7. 7.
    Assenmacher M, Schmitz J, Radbruch A (1994) Flow cytometric determination of cytokines in activated murine T helper lymphocytes: expression of interleukin-10 in interferon-gamma and in interleukin-4-expressing cells. Eur J Immunol 24:1097–1101CrossRefGoogle Scholar
  8. 8.
    Elson LH, Nutman TB, Metcalfe DD et al (1995) Flow cytometric analysis for cytokine production identifies T helper 1, T helper 2, and T helper 0 cells within the human CD4+CD27− lymphocyte subpopulation. J Immunol 154:4294–4301PubMedGoogle Scholar
  9. 9.
    Jung T, Schauer U, Heusser C et al (1993) Detection of intracellular cytokines by flow cytometry. J Immunol Methods 159:197–207CrossRefGoogle Scholar
  10. 10.
    Prussin C, Metcalfe DD (1995) Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods 188:117–128CrossRefGoogle Scholar
  11. 11.
    Sander B, Hoiden I, Andersson U et al (1993) Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. Cytokine detection by immunoassay and intracellular immunostaining. J Immunol Methods 166:201–214CrossRefGoogle Scholar
  12. 12.
    Carlyle JR, Mesci A, Ljutic B et al (2006) Molecular and genetic basis for strain-dependent NK1.1 alloreactivity of mouse NK cells. J Immunol 176:7511–7524CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of ImmunologyIRCSS Bambino Gesù Children’s HospitalRomeItaly
  2. 2.Aix Marseille University, CNRS, INSERM, Centre d’Immunologie de Marseille-LuminyMarseilleFrance

Personalised recommendations