FIG 8.

HLA-A*0201-restricted polyfunctional CD8⁺ T cells elicited by MVA-7Ag1. HLA-A*0201 (A2) transgenic mice were immunized two times in 4-week intervals with MVA-7Ag1, MVA^B-pp65 (n = 5), or MVA-Venus (n = 1). One week postimmunization, antigen-specific T cell responses were evaluated by multicytokine ICS following stimulation with pp65- and PC-specific peptide libraries or an HLA-A*0201-restricted immunodominant pp65 peptide epitope. (A to C) Shown are the percentages of CD8⁺ T cells secreting IFN-γ (A), TNF-α (B), or IL-2 (C) following stimulation of splenocytes from A2 immunized mice with different stimuli. (D to F) Frequency of antigen-specific CD8⁺ T cells producing all combinations of IFN-γ, TNF-α, and IL-2 cytokines following in vitro stimulation with the HLA-A*0201-restricted peptide epitope (D), pp65-specific peptide library (E), or PC-specific peptide library (F) in MVA-7Ag1-, MVA^B-pp65-, or MVA-Venus-immunized B7 mice. Horizontal bars represent median values. (G) Pie charts show the relative contribution of each polyfunctional subset among the total polyfunctional response to the pp65 epitope, pp65 library, or PC library in MVA-7Ag1- or MVA^B-pp65-immunized A2 mice. Each pie chart represents the mean response across the immunized mice to the three different antigen stimulations. The average total percentage of CD8⁺ T cells responding to the peptide stimulation is shown under each pie chart. Polyfunctional subsets and functions (Fn°) are indicated in the color key. Significance of the difference between the groups was calculated using multiple t test.