Figure 8.

Analysis of CD8+ T-cell multifunctionality. PBMCs were incubated with pools of HPV peptides for 10 days, re-stimulated with HPV E7 22-mer peptides, stained with antibodies recognizing various surface and functional markers (CD3, CD4, CD8, IFN-γ, TNF-α, IL-2, CD154), and analysed by flow cytometry as described under Materials and Methods. (A) Assessment of the co-expression of CD154, IFN-γ, IL-2, and TNF-α. The pie charts illustrate the relative representation of cells co-expressing all four, different combinations of three, different combinations of two, or only a single of these functional markers within total functional cells (i.e. all cells that are positive for CD154 and/or IFN-γ and/or IL-2 and/or TNF), for those samples that had measurable CD8+ T-cell responses to HPV16 E7 peptide re-stimulation as defined in Materials and Methods. The data are not background-subtracted. (B) Percentage of CD8-cells expressing multiple (≥3) functional markers. Results are shown for each subject defined as a responder under the criteria described in Materials and Methods.