In vivo adjuvant activity of in vitro–generated Th1-like NKT cells. Th1-like NKT cells were generated as shown in Figure 2C. Jα18^−/− mice were reconstituted with or without (No ES-NKT) Th1-like NKT cells (2 × 10⁶/mouse) and immunized with OVA-loaded TAP^−/− spleen cells (2 × 10⁷) and α-GalCer (2 μg; TOG immunization). One week later, spleen cells were restimulated in vitro with or without 1μM OVA peptide (257-264) as described.³⁴  As a control, spleen cells from wild-type B6 mice or Jα18^−/− mice reconstituted with Th2-like NKT cells (generated as shown in Figure 2B and that had been treated with or without TOG immunization) were stimulated in vitro with or without OVA_257-264 peptide. The percentage of IFN-γ–producing CD8⁺ T cells stimulated with or without OVA is shown by the intracellular cytokine staining (boxed). One representative of 4 independent experiments is shown.