Abstract
Innate lymphoid cells (ILCs) are present in most tissues within the body but have been most extensively studied within mucosal barriers such as the lung and intestine. Isolation of immune cells from such tissues requires enzymatic digestion, and the number and composition of the cells released are dependent upon robust protocols tailored to the tissue of study. Here, detailed methodologies to isolate ILCs from various barrier sites and their draining lymph nodes (LNs) are described. Flow cytometry staining and gating strategies for identification and quantification of ILCs are then provided. Combined, these provide an efficient means to study ILCs within the small intestine lamina propria, lung, ear skin, and LNs.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Moro K, Yamada T, Tanabe M et al (2010) Innate production of T(H)2 cytokines by adipose tissue-associated c-Kit(+)Sca-1(+) lymphoid cells. Nature 463:540–544
Neill DR, Wong SH, Bellosi A et al (2010) Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity. Nature 464:1367–1370
Sawa S, Cherrier M, Lochner M et al (2010) Lineage relationship analysis of RORgammat+ innate lymphoid cells. Science 330:665–669
Artis D, Spits H (2015) The biology of innate lymphoid cells. Nature 517:293–301
Serafini N, Vosshenrich CA, Di Santo JP (2015) Transcriptional regulation of innate lymphoid cell fate. Nat Rev Immunol 15:415–428
Spits H, Artis D, Colonna M et al (2013) Innate lymphoid cells—a proposal for uniform nomenclature. Nat Rev Immunol 13:145–149
Walker JA, Barlow JL, Mckenzie AN (2013) Innate lymphoid cells—how did we miss them? Nat Rev Immunol 13:75–87
Klose CS, Flach M, Mohle L et al (2014) Differentiation of type 1 ILCs from a common progenitor to all helper-like innate lymphoid cell lineages. Cell 157:340–356
Gordon SM, Chaix J, Rupp LJ et al (2012) The transcription factors T-bet and Eomes control key checkpoints of natural killer cell maturation. Immunity 36:55–67
Halim TY, Krauss RH, Sun AC et al (2012) Lung natural helper cells are a critical source of Th2 cell-type cytokines in protease allergen-induced airway inflammation. Immunity 36:451–463
Price AE, Liang HE, Sullivan BM et al (2010) Systemically dispersed innate IL-13-expressing cells in type 2 immunity. Proc Natl Acad Sci U S A 107:11489–11494
Wilhelm C, Hirota K, Stieglitz B et al (2011) An IL-9 fate reporter demonstrates the induction of an innate IL-9 response in lung inflammation. Nat Immunol 12:1071–1077
Yagi R, Zhong C, Northrup DL et al (2014) The transcription factor GATA3 is critical for the development of all IL-7Ralpha-expressing innate lymphoid cells. Immunity 40:378–388
Halim TY, Maclaren A, Romanish MT et al (2012) Retinoic-acid-receptor-related orphan nuclear receptor alpha is required for natural helper cell development and allergic inflammation. Immunity 37:463–474
Hoyler T, Klose CS, Souabni A et al (2012) The transcription factor GATA-3 controls cell fate and maintenance of type 2 innate lymphoid cells. Immunity 37:634–648
Walker JA, Oliphant CJ, Englezakis A et al (2015) Bcl11b is essential for group 2 innate lymphoid cell development. J Exp Med 212:875–882
Cherrier M, Sawa S, Eberl G (2012) Notch, Id2, and RORgammat sequentially orchestrate the fetal development of lymphoid tissue inducer cells. J Exp Med 209:729–740
Eberl G, Marmon S, Sunshine MJ et al (2004) An essential function for the nuclear receptor RORgamma(t) in the generation of fetal lymphoid tissue inducer cells. Nat Immunol 5:64–73
Sonnenberg GF, Monticelli LA, Alenghat T et al (2012) Innate lymphoid cells promote anatomical containment of lymphoid-resident commensal bacteria. Science 336:1321–1325
Takatori H, Kanno Y, Watford WT et al (2009) Lymphoid tissue inducer-like cells are an innate source of IL-17 and IL-22. J Exp Med 206:35–41
Mebius RE (2003) Organogenesis of lymphoid tissues. Nat Rev Immunol 3:292–303
Hepworth MR, Fung TC, Masur SH et al (2015) Immune tolerance. Group 3 innate lymphoid cells mediate intestinal selection of commensal bacteria-specific CD4(+) T cells. Science 348:1031–1035
Hepworth MR, Monticelli LA, Fung TC et al (2013) Innate lymphoid cells regulate CD4+ T-cell responses to intestinal commensal bacteria. Nature 498:113–117
Bernink JH, Krabbendam L, Germar K et al (2015) Interleukin-12 and -23 control plasticity of CD127(+) group 1 and group 3 innate lymphoid cells in the intestinal lamina propria. Immunity 43:146–160
Bernink JH, Peters CP, Munneke M et al (2013) Human type 1 innate lymphoid cells accumulate in inflamed mucosal tissues. Nat Immunol 14:221–229
Klose CS, Kiss EA, Schwierzeck V et al (2013) A T-bet gradient controls the fate and function of CCR6-RORgammat+ innate lymphoid cells. Nature 494:261–265
Dutton EE, Camelo A, Sleeman M et al (2017) Characterisation of innate lymphoid cell populations at different sites in mice with defective T cell immunity. Wellcome Open Res 2:117
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Dutton, E.E., Withers, D.R. (2020). Identification of Murine Innate Lymphoid Cell Subsets in Barrier Tissues and Their Draining Lymph Nodes Using Flow Cytometry. In: Amarnath, S. (eds) Innate Lymphoid Cells . Methods in Molecular Biology, vol 2121. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0338-3_3
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0338-3_3
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0337-6
Online ISBN: 978-1-0716-0338-3
eBook Packages: Springer Protocols