Figure 5.

Vaccine-elicited cytokine production is due to different effector/memory phenotypes based upon culture conditions. For long-term culture, PBMCs were isolated at 6 WPV and stimulated with a cocktail of Ag85A, TB10.4, and PPD for 13 days followed by transfer to 96-well round bottom plates with APCs and addition of media alone PPD (A) or Ag85A/TB10.4 (B) for an additional 16 h. Cytokine production and effector/memory phenotype were determined by ICS flow cytometry. Results are presented as average and SE. Frequency of cytokine-producing cells under long-term or ex vivo culture exhibiting Tcm, Tem, effector, or naïve phenotype using the gating strategy described in Figure S1 in Supplementary Material. *Differences (P < 0.05) in cytokine production within memory subsets (i.e., Tcm, Tem, effector, or naïve) in long-term as compared to ex vivo cultures (Tukey’s).