Strong polyfunctional CD4+ and CD8+ T cell responses directed against Rv0477c
target peptides. PMBCs were obtained after BCG vaccination and tested by ICS for IFN-γ, TNF-α and IL-2 production in CD4+ and CD8+ T cells. First, cells were gated on site and forward scatter, followed by gating on CD3+ events and CD4+ versus CD8+ T-cells. Responding T-cells were first tested for IFN-γ production, followed by IL-2 versus TNF-α production in the IFN- γ + and IFN-γ - T cell populations. Controls included stimulation with medium or with PMA/ionomycin (data not shown).