Culture and transduction of human iNKT cells. (A) iNKT cells in PBMCs were expanded 300-folds after stimulation with α-GalCer for 14 d. The upper panel: prominent cell colonies were observed (Magnification: 200×); the lower panel: the percentage of iNKT cells in PBMCs and the purity of iNKT cells after isolation with microbeads. (B) The purity of iNKT cells before and after isolation with microbeads, (C) and the expression of the PE-Vα24 (red) and APC-Vβ5 (blue) genes after retroviral transduction, were observed under fluorescence microscopy (D) and analyzed with Flow cytometry. (E) Intracellular Flow cytometry analysis the iNKT cell populations expressed IFN-γ upon activated by Mtb antigen 38-kDa stimulation. UnTd (untransduced iNKT cells); EmTd (empty vector-transduced iNKT cells); Td (TCR gene-modified iNKT cells). Td + 38-kDa (TCR gene-modified iNKT cells + 38-kDa antigen-loaded DCs).