Fig. 5.

Analysis of T cell receptor (TCR) Vα24/Vβ11 expression and CD1d tetramer staining of natural killer (NK) T cell lines. NK T cell lines were established by fluorescence activated cell sorting of anti-TCR Vα24 plus Vβ11-labelled cells and cultured for 1–3 weeks in medium supplemented with interleukin (IL)-2 and IL-15 (see Materials and methods). Cells double-stained with anti-TCR Vα24- fluorescein isothiocyanate (FITC) and Vβ11-phycoerythrin (PE) are detected within lymphocyte and CD3 gate; for CD1d tetramer staining, cells were stained with PE-conjugated PBS57 (αGalCer homologue) loaded CD1d tetramer combined with negative selection antibodies (a mixture of FITC-conjugated anti-CD13, 14, 16, 19). The percentages of CD1d tetramer binding T cells are shown in the upper left quadrants.