Isolation and analysis of group 2 innate lymphoid cells in mice

Kazuyo Moro, Kafi N. Ealey, Hiroki Kabata, Shigeo Koyasu

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Recent studies have identified distinct subsets of innate lymphocytes, collectively called innate lymphoid cells (ILCs), which lack antigen receptor expression but produce various effector cytokines. Group 2 ILCs (ILC2s) respond to epithelial cell-derived cytokines such as interleukin (IL)-25, IL-33 and thymic stromal lymphopoietin (TSLP), produce large amounts of type 2 cytokines, and have a key role in anti-helminth innate immunity and in the pathophysiology of allergic inflammation. The reported phenotypic characteristics of mouse ILC2s vary, depending on the tissue source and preparation method. This protocol describes improved methods for tissue-specific isolation and analysis of mouse ILC2s of high purity and yield from fat tissue, lung, bronchoalveolar lavage fluid (BALF) and small intestine. These improved methods are the result of our thorough investigation of enzymes used for tissue digestion, methods for the elimination of undesired cells, and a combination of antibodies for the detection and isolation of ILC2s. In addition, this new protocol now enables the isolation of ILC2s of high yield, even from inflamed tissues. Depending on the tissue being analyzed, it takes ∼ 2-4 h for isolation and flow cytometric analysis of ILC2s from the various tissues of a single mouse and ∼ 4-8 h to sort purified ILC2s from pooled tissues of multiple mice.

Original languageEnglish
Pages (from-to)792-806
Number of pages15
JournalNature Protocols
Volume10
Issue number5
DOIs
Publication statusPublished - 2015 May 6

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Lymphocytes
Tissue
Bronchoalveolar Lavage Fluid
Cytokines
Antigen Receptors
Interleukin-17
Helminths
Lymphocyte Subsets
Innate Immunity
Small Intestine
Digestion
Epithelial Cells
Fats
Inflammation
Fluids
Antibodies
Enzymes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Isolation and analysis of group 2 innate lymphoid cells in mice. / Moro, Kazuyo; Ealey, Kafi N.; Kabata, Hiroki; Koyasu, Shigeo.

In: Nature Protocols, Vol. 10, No. 5, 06.05.2015, p. 792-806.

Research output: Contribution to journalArticle

Moro, Kazuyo ; Ealey, Kafi N. ; Kabata, Hiroki ; Koyasu, Shigeo. / Isolation and analysis of group 2 innate lymphoid cells in mice. In: Nature Protocols. 2015 ; Vol. 10, No. 5. pp. 792-806.
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