Isolation and Characterization of Conventional and Non-conventional Type 2 Innate Lymphoid Cells (ILC2s) from Human Peripheral Blood Mononuclear Cells (PBMCs)

AbstractInnate 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.Key wordsInnate lymphoid cellsPeripheral blood mononuclear cellsFlow cytometryCRTH2 (Chemoattractant Receptor-Homologous Molecule Expressed on Th2 cells)CD30TNFR2 (Tumor Necrosis Factor Receptor 2)