Figure 2

Gating strategy to identify immune cell populations in different swine tissues.

Single cell suspensions were labeled with PBS57-loaded and unloaded CD1d-tetetramer and various antibodies. (a) T cells were identified as CD3⁺ cells after first gating on single-celled, live lymphocytes according to size and granularity. NKT-cells were distinguished from conventional T cells according to CD1d-tetramer staining. NKT-cell and conventional T cell subsets were distinguished according to surface expression of CD4, CD8α, CD44, and CD11b. (b) NK cells were identified as CD8α⁺ CD3⁻ cells after gating single live lymphocytes. Different NK cell subsets were distinguished from each other according to Nkp46 and CD11b surface expression (c) γδ T cells were identified as TCRδ⁺ CD3⁺ cells after gating on single-celled, live lymphocytes. (d) Monocytes and granulocytes were identified as CD172α⁺ cells with respectively high and low granularity according to side scatter. One representative lung sample is shown.