The innate lymphoid cells (ILCs) family comprise a group of newly discovered cytokine-secreting cells with lymphoid morphology that lack rearranged antigen-specific receptors. ILCs play an important role in many immune processes, including control of infections, inflammation, and tissue repair. ILCs are divided into three groups, ILC1, ILC2, and ILC3. ILC2 are critical for the initiation of anti-helminthic and allergic immune responses producing high levels of Tʜ2 cytokines IL-5 and IL-13.
Human type 2 innate lymphoid cells (ILC2) are defined as lineage (CD2, CD3, CD14, CD16, CD19, CD56, CD235a, CD123)-negative and (CD127, CD161, CRTH2)-positive lymphocytes.1 Using the ILC2 Isolation Kit, human type 2 innate lymphoid cells are pre-enriched in a first step by depletion of lineage-positive cells, to facilitate the enrichment of highly pure target cells through subsequent enrichment using CD294 (CRTH2)-PE and Anti-PE MicroBeads.
Human type 2 innate lymphoid cells (ILC2) isolated from human peripheral blood mononuclear cells (PBMCs) can be used for further phenotypical or functional characterization.
Type 2 innate lymphoid cells (ILC2) were isolated from peripheral blood mononuclear cells by using the ILC2 Isolation Kit, two LS and two MS Columns, as well as a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD45-VioBlue® and lineage markers (CD2, CD3, CD14, CD16, CD123, CD235a (Glycophorin A) conjugated to FITC, CD19 and CD56 conjugated to VioBright™ FITC), and CD294 (CRTH2)-PE (A) or CD161-PE-Vio® 770 and CD127-APC (B) and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals, CD45, and propidium iodide fluorescence.
Isolated CD294 (CRTH2)+ ILC2