Figure 3.

Ex vivo expansion of human iNKT cells by α-GlcCer and α-GalCer analogues. Peripheral blood mononuclear cells (PBMC) from four different donors were stimulated with the indicated glycolipids at a concentration of 250 nM in the presence of low levels of exogenous IL-2 and IL-7. At day 8, cultures were harvested and analysed by flow cytometry using monoclonal antibodies specific for CD3 and for the invariant TCRα chain expressed by iNKT cells (6B11). (A) Dot plots showing relative levels of CD3⁺ 6B11⁺iNKT cells are shown for one representative donor. Numbers in upper right quadrant indicate percentages of total lymphocytes that are iNKT cells. (B) Absolute numbers of iNKT cells in the cultures were determined by flow cytometry using fluorescent counting beads, and the values of iNKT cell fold expansion were determined by dividing by the input number of iNKT cells.