Fig. 4

Similar frequencies of T cells recognizing Mtb-infected macrophages are detected by the MIME assay and MIM-ICS. a Pulmonary T cells were cultured with Mtb-infected macrophages and analyzed by ICS and flow cytometry. Representative flow plots showing the frequency of pulmonary CD4 (left column) or CD8 (right column) T cells expressing CD69 and producing IFN-γ after a 6 h culture with Mtb-infected macrophages. b The MIME or the Mtb-infected macrophage-ICS (MIM-ICS) assay were used to calculate the frequency of CD4 (left) or CD8 (right) pulmonary T cells that recognized Mtb-infected macrophages. U uninfected macrophages, Mtb Mtb-infected macrophages. n.s. not significant (t test). c The frequency of ESAT-6- or Ag85b-specific CD4 T cells (left) or the frequency of TB10.4- or 32a-specific CD8 T cells (right) among T cells from the lungs of Mtb-infected mice as determined by elispot or ICS, using peptide-pulsed uninfected macrophages, or tetramer staining. Data are representative of two independent experiments using pooled T cells from five mice at 5 WPI (shown, a–c) or 7.5 months post infection, analyzed in triplicates. Ordinary one-way ANOVA with Tukey’s multiple comparisons was performed for statistical analyses, adjusted p values: *≤0.05, **≤0.01, ***≤0.001, ****≤0.0001