Flow cytometric analysis of innate lymphoid cells: challenges and solutions

Introduction The three groups of helper innate lymphoid cells (ILCs), namely ILC1, ILC2 and ILC3, have been identified by flow cytometry combinations cell surface markers. Here, we review various ways ILCs are currently identified, focusing on potential problems their solutions. first step to identify all is exclude other lymphocytes myeloid lineage-specific markers (Lin). However, the Lin cocktail varies in studies, definition Lin- population containing often ambiguous, resulting contamination + cells, particularly T cells. Method We designed ILC populations tissues B6 mice cytometry. To minimize contamination, TCR/CD3ϵ antibodies were used separately from cocktail. confirmed expression transcription factors GATA3, RORγt, T-bet Eomes. Result ILC1s mouse liver Lin-NKp46 NK1.1 − CD49a CD49b . defining remains a challenge. ILC2s lung Thy1 CD127 ST2 whereas small intestine GATA3 RORγt ILC3s spleen, liver, Discussion heterogeneous strategies be for each tissue. Excluding cases crucial, combination T-bet, Eomes should ILCs. Using CD3ϵ/TCRs different fluorochrome not minimizes specifically individual tissues.