Induction of IFN-γ- and TNF-α-secreting CD8⁺ T cell subpopulations upon DC priming.

<p>PBMC from volunteer CVD4000#65 were co-cultured with DC alone (media), or pre-mixed with live or heat-killed <i>S.</i> Typhi at a MOI of 10∶1, or uninfected or <i>S.</i> Typhi-infected cells at a 1∶4 blast∶DC ratio. After 20 hours of incubation, cells were surface stained with a combination of mAb to CD3, CD4, CD8, CD14, CD19, CD45RA, and CD62L as well as ViViD. After fixation and permeabilization, cells were intracellularly stained for IFN-γ and TNF-α and analyzed by multichromatic flow cytometry. Lymphocytes gated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005879#pone-0005879-g005" target="_blank">Figure 5</a> were followed by additional gating on CD3, CD4 and CD8, as well as CDR45RA versus CD62L to analyze T_EM and T_EMRA cell subsets. Numbers correspond to the percentage of positive cells in the indicated regions in each histogram. These results are representative of 1 of 4 volunteers with similar results.