Figure 4
Phenotypic and functional assessment of virus-specific CD8
+ T cells in YATECs.
(A) Representative staining of CD27 on HLA-A11 EBV EBNA-3B AK10–specific CD8+ T cells identified in 1 YATEC and 1 control subject. Percentages of CD27– or CD27+ cells within tetramer-positive populations are shown. (B) Expression of the differentiation markers CD27 or CD57 on EBV- or CMV-specific CD8+ T cells, identified using tetramers in YATECs or control individuals. Horizontal bars indicate the median. (C) Representative example of simultaneous multifunctional assessment of B8-FL9 EBV–specific CD8+ T cells from a YATEC patient. Cells were stimulated overnight in the presence of cognate peptide prior to intracellular staining and analysis using 9-color flow cytometry. Percentages of cells in the different quadrants are shown. Plots are gated on CD3+CD8+ T cells and tetramer-positive T cells, respectively. (D) Polyfunctional profiling of EBV-specific CD8+ T cell populations from YATECs (n = 8) or controls (n = 10) upon stimulation with their cognate peptide. For simplicity, percentages of cells are grouped according to the number of functions (from CD107a or CD40L, IFN-γ, TNF-α, IL-2, and MIP-1β) elicited in response to antigen encounter. Individual segments of the pie charts represent the proportions of cells within each total population that exhibited the number of functions indicated. (E) Polyfunctional profiling of CMV-specific CD8+ or CD4+ T cell populations from CMV-seropositive YATECs (n = 15) or CMV-seropositive controls (n = 15) upon stimulation with pp65 and IE1 overlapping peptides.
