Figure 3.

Poly-functional CD4⁺ T cells are boosted following ZOSTAVAX vaccination. PBMC were thawed and rested overnight before VZV antigen stimulation. One million PBMC/sample were stimulated with antigens for 5.5 h at 37°C/5% CO₂, with brefeldin A and monensin added 30 min after the initial incubation. Cells were stained with Vividye, then fixed, permeabilized, and stained with antibodies to identify (A) CD3⁺ T cells, expressing either CD4 or CD8 that were producing (B) IFNγ, IL-2, TNFα, and CD154. Figure shows IE62-specific responses from one individual, and numbers on dot plots represent percentages of cells (DMSO subtracted) expressing each molecule. (C) Shows the heterogeneity of IE63-specific poly-functional CD4⁺ T cells for all 21 subjects as determined by Boolean-gating strategy. Each dot represents one subject, and the mean is shown for all phenotypes.