Effect of LIGHT reverse signaling on CTL development.
2C mouse spleen cells (4 × 106 cells/2 mL/well) were mixed with an equal amount of mitomycin C– treated BALB/c mouse spleen cells (4 × 106cells/2 mL/well) and seeded in flat-bottomed 24-well plates, which were precoated with TR6-FC (10 μg/mL) or normal human IgG (NhIgG, 10 μg/mL) as a control. Soluble human LIGHT (20 μg/mL) was added at the beginning of the culture in certain samples as indicated. The cells were cultured in the presence of 10 U/mL IL-2 for 6 days. CTL activity in the stimulated cells was then measured by a standard 4-hour51Cr-release assay, using P815 cells as targets. The samples were tested in triplicate, and means ± SD of percentage of target cell lysis are shown. The experiments were performed twice with similar results, and the data of a representative experiment are presented.
