Figure 3. γδ NKT cells in the thymus of Itk^−/− mice have characteristics of less mature αβ iNKT cells.

Thymocytes from WT and Itk^−/− mice, or WT-4Get and Itk^−/−-4Get mice were stained and analyzed by flow cytometry.

(A) Total thymocytes were analyzed for TCR versus TCRδ (left panels), and TCRδ-positive cells were gated on and examined for Vγ1.1 and Vδ6.3 expression (second panels). V6 cells were further gated on HSA (third panels). iNKT cells were identified by staining with CD1d tetramer and anti-TCR² (fourth panels). iNKT cells were further gated on HSA expression (right panels).

(B) HSA⁻ V6 γδ T cells and iNKT cells in the thymus were analyzed for expression of CD44, CD122, CD4 and intracellular PLZF. Dot-plots show representative data; compilations of data from of all experiments are shown in the graphs below. Statistically significant differences are shown with p values.

(C) HSA⁻ V6 γδ T cells and iNKT cells from WT-4Get and Itk^−/−-4Get mice were stained and analyzed for CD44 vs GFP, CD122 vs GFP and CD4 vs GFP expression. Data are representative of at least two independent experiments.

(D) PLZF^hi and PLZF^int V6 γδ T cell and iNKT cell populations were gated as shown in panel B and absolute numbers were calculated (left two panels). The percentages and absolute numbers of GFP⁺ cells in the HSA⁻ V6 γδ T cell and iNKT cell populations are shown (right two panels). Statistically significant differences are shown with p values.