Innate Lymphoid Cell Gating Strategies for Flow Cytometry

Flow cytometry is essential for the study of innate lymphoid cells (ILCs). It allows multiparameter analysis of cell populations based on the expression of specific cell surface and/or intracellular molecules. Consensus gating strategies were established in collaboration with researchers in the ILCs field.1,2 For ILC single cell suspension preparation, R&D systems scientists used previously established protocols.3,4

Group 1 ILCs

Natural killer cells
Mouse  CD3-CD49b+CD161+NKp46+CD27high/low
Human CD3- CD57+CD56dim/brightCD16dim/-
Mouse/Human CD34- CD127+NKp44-CD117-
Mouse  CD45+CD3 epsilon-CD19-CD161+NKp46+CD160+
Human  CD45+CD3 epsilon-CD19-NKp44+NKp46+CD103+

Group 2 ILCs

Mouse/Human  CD45+CD3 epsilon-CD4-CD8-CD19-CD11b-CD161-CD90.2+T1/ST2+GATA3high

Group 3 ILCs

LTi cells
Mouse/Human CD4+/-CD127+ Integrin alpha 4 beta 7+ROR gamma t+NKp46-CCR6+
Mouse  CD45+CD3-CD19-ROR gamma t+NKp46-CD161 T-bet+/-
Human CD45+CD3-CD19-CD127+NKp44+CCR6-T-bet+/-
Mouse  CD45+CD3-CD19-ROR gamma t+NKp46+CD161low/- T-bethigh
Human  CD45+CD3-CD19-ROR gamma t+CD127+NKp44+CCR6+T-bethigh

More ILC Resources:

R&D Systems Innate Lymphoid Cells Differentiation Pathway

R&D Systems optimized protocols for Flow Cytometry samples preparation

  1. Fuchs, A. et al (2013) Immunity. 38:769
  2. Molofsky, A.B. et al (2013) J. Exp. Med. 210:535
  3. Morita, R. et al (2011) Immunity. 34: 108
  4. Hepworth, M.R. et al (2013) Nature. 498 :113


    Detection of CCR2 in Mouse Peripheral Blood Cells by Flow Cytometry. Mouse peripheral blood cells were stained with an APC-conjugated Rat Anti-Mouse Integrin alpha M/CD11b Monoclonal Antibody (Catalog # FAB1124A) and either (A) a PE-conjugated Rat Anti-Mouse/Rat CCR2 Monoclonal Antibody (Catalog # FAB5538P) or (B) an APC-conjugated Rat IgG2B Isotype Control (Catalog # IC013A).