Confirmation of <i>H-2^b</i>-restricted T cell epitopes in MPT83.

<p>C57BL/6 mice (n = 3) were immunized with 100 µg of DNA-MPT83 or pcDNA3 control vector by i.m.i three times at two-weekly intervals. Four weeks post-immunization, mice were harvested, and the pooled splenocytes were labelled with anti-CD3-PerCP, anti-CD4-Alexaflour 700 and anti-CD8-APC-Cy7 mAbs and the CD4⁺ and CD8⁺ T cells were separated by flow cytometry. The CD4⁺ (A) and CD8⁺ (B) T cells were stimulated with irradiated splenocytes and the peptides 1, 5, 9, 13 and 14 (10 µg/ml), MPT83 (10 µg/ml) or ConA (3 µg/ml), and the antigen-specific IFN-γ responses were measured by ELIspot after 16 h. Data represent the means of replicates for each treatment, and are representative of two independent experiments.