Protocol for Determining the Effect of Neuroendocrine Hormones on Murine ILC Function
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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 wordsNK cells ILCs Glucocorticoids Flow cytometry IFN-γ Cytotoxic granule exocytosis
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.