Isolation and Identification of Group 2 Innate Lymphoid Cells in Settings of Type 2 Inflammation

: The recent discovery of innate lymphoid cells (ILCs), a major source of canonical T cell cytokines, has prompted significant interest into understanding the role of these novel cells in immune responses. Unlike T cells, ILCs lack antigen receptors, instead receiving activation from locally derived tissue signals. Group 2 ILCs (ILC2s), which express the genes encoding GATA-3, interleukin (IL)-4, IL-5, IL-9, and IL-13, are distributed throughout nonlymphoid tissues. Although ILC2s lack antigen receptors, there are considerable similarities that they share with their Th2 cell counterparts, including receptors, secreted signals, and key transcription factors. Here we describe a method of isolating ILC2s for analysis from peripheral tissues of the mouse. The approach consists of digesting and mechanically dissociating harvested organs followed by staining with fluorescently labeled antibodies for flow cytometry or cell sorting. We suggest panels of antibodies for each tissue that can be used as positive and negative markers to selectively separate ILC2s from other cells, and we demonstrate marker specificity with example cells from a "cytokine reporter" mouse strain.