Innate lymphoid cells - Flow cytometry for ILCs

4 important questions on Innate lymphoid cells - Flow cytometry for ILCs

What does histopaque do in blood samples?

In density gradient centrifugation, histopaque is a solution that allows blood cells to separate based on how heavy they are. Leads to PBMC, lymphocyte layer.

What are the axes of flow cytometry graphs?

X axis: Size of the cell
Y axis: cell granularity

How can we find different ILCs with flow cytometry?

1. Extract PBMC with histopaque
2. Culture the contents of PBMC
3. Get rid of everything else that is not lymphocytes (e.g. RBC, granulocytes)
4. Get rid of dead cells
5. Use Lin- to get rid of differentiated cells that are not ILCs
6. Use CD45+ to get rid of other non immune cells
5. Use CD127+ to find ILCs, it is the receptor marker for IL-7, which is a survival marker for ILC
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What are the master transcription factors for each ILC?

ILC1: T-bet, for its identity
ILC2: GATA3, for its identity
ILC3: RORyt (RORC), for its differentiation

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