Figure 4.

IFN-γ treatment in combination with BCG or BCG alone induces similar proportions of antigen-specific Vβ17⁺ T cells in recall responses, but which differ in IFN-γ producing ability DC were loaded with inactivated Influenza A virus or control protein (KLH) by pulsing with these proteins for 1 h followed by 2 hours of incubation before maturation with BCG (1.6 × 10⁶ CFU/ml), BCG and IFN-γ (1000 U/ml), or left untreated (immature). After 20 hours of maturation, the DC were washed and autologous PBMC were added at a 10:1 (PBMC:DC) ratio in AIM-V media containing IL-2 and IL-15. Seven or eight days later the resulting cell lines were counted, analyzed for the percentage of Vβ17⁺CD8⁺ expression on T cells by flow cytometry, and the results calculated as a percent of the response seen using Influenza A-pulsed immature DC as the control (A). The influenza A virus-specific Vβ17⁺CD8⁺ T cells were re-stimulated for 40 h (including addition of GolgiPlug™ in the last 16 h) with immature or BCG ± IFN-γ matured DC loaded with Influenza M1 protein and evaluated for intracellular IFN-γ (B). Data in panel A are combined from three separate experiments and data in panel B are from one representative experiment out of four similar experiments.