Abstract
Innate lymphoid cells (ILCs) are a relatively new family of lymphoid cells that lack lineage cell surface markers but produce various effector cytokines. Based on phenotype and function, the group 2 ILCs (ILC2s) mirror the features of the adaptive CD4+ Th2 cell subset. In humans, they are traditionally characterized as the Lin−IL7Rα+CRΤΗ2+CD161+ cell population that produces type 2 cytokines – IL-5 and IL-13. However, the commonly used surface markers for human ILC2s leave a majority of type 2 cytokine-producing ILC2s unaccounted for. Recently, we characterized a distinct type 2 cytokine-producing Lin− population that lacks surface expression of canonical CRTH2 but expresses CD30 and TNFR2. Herein, we describe a detailed protocol for isolation, staining, and analysis of the conventional Lin−CRTH2+IL7Ra+ and the non-conventional Lin−CD30+TNFR2+ ILC2 populations.
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
Similar content being viewed by others
References
Klose CSN, Artis D (2020) Innate lymphoid cells control signaling circuits to regulate tissue-specific immunity. Cell Res 30:475–491
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
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
Mjösberg JM, Trifari S, Crellin NK et al (2011) Human IL-25- and IL-33-responsive type 2 innate lymphoid cells are defined by expression of CRTH2 and CD161. Nat Immunol 12:1055–1062
Björklund ÅK, Forkel M, Picelli S et al (2016) The heterogeneity of human CD127(+) innate lymphoid cells revealed by single-cell RNA sequencing. Nat Immunol 17:451–460
Kirkham CL, Carlyle JR (2014) Complexity and diversity of the NKR-P1:Clr (Klrb1:Clec2) recognition systems. Front Immunol 5:214
De Fanis U, Mori F, Kurnat RJ et al (2007) GATA3 up-regulation associated with surface expression of CD294/CRTH2: a unique feature of human Th cells. Blood 109(10):4343–4350
Cosmi L, Annunziato F, Galli MIG et al (2000) CRTH2 is the most reliable marker for the detection of circulating human type 2 Th and type 2 T cytotoxic cells in health and disease. Eur J Immunol 30:2972–2979
Liu S, Sirohi K, Verma M et al (2020) Optimal identification of human conventional and nonconventional (CRTH2-IL7Rα-) ILC2s using additional surface markers. J Allergy Clin Immunol 146:390–405
Acknowledgments
The work was supported by NIH grants AI102943, AI137970, and HL126895; and grants from the Cohen Family Foundation, NBL Fellowship Foundation, Judy Renick Research Fund and Colorado Technology and Department of Medicine of NJH.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Sripada, A., Sirohi, K., Alam, R. (2022). Isolation and Characterization of Conventional and Non-conventional Type 2 Innate Lymphoid Cells (ILC2s) from Human Peripheral Blood Mononuclear Cells (PBMCs). In: Gorska, M.M. (eds) Asthma. Methods in Molecular Biology, vol 2506. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2364-0_13
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2364-0_13
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2363-3
Online ISBN: 978-1-0716-2364-0
eBook Packages: Springer Protocols