Fig. 4

Multifunctional analysis of CD4⁺ and CD8⁺ T cell cytokine responses to RSV. Day 8 and day 29 boolean ‘OR’ gates for CD4⁺ and CD8⁺ T cell IFNγ⁺, TNFα⁺, IL-2⁺, IL-4⁺ and IL-17⁺ expression in response to RSV (F) peptide stimulation, were generated in Flowjo 10.1. The average of the boolean ‘OR’ gates was used to determine the top three responders. Multifunctional analysis revealed heterogeneity in cytokine production among the subjects as well as dominance of single cytokine production both pre- and post-vaccination compared to dual- or tri-CD4⁺ (a) and CD8⁺ (b) positive cytokines