Fig. 5

Expansion of antigen-specific T cells in a CMV model system. Monocytes and T cells were isolated from peripheral blood mononuclear cells of donors screened for having CMV-specific T cells. Monocytes were differentiated with GM-CSF/IL-4 to immature dendritic cells (DCs). DCs were infected with LOAd(-), LOAd703 or LOAd732, or stimulated with Poly(I:C)/TNFα (positive control), or left untreated. 24 h later, DCs were pulsed with the CMV peptide pp65 and co-cultured with autologous T cells. After 11 days of co-culture, cells were harvested and analyzed with flow cytometry. In A, the percentage of expanded CMV-specific T cells (CD3 + CD8 + CMVtet +), natural killer (NK) cells (CD3-CD56 + /CD16 +) and regulatory T cells (Tregs: CD3 + CD4 + CD25 + CD127low/-) is shown (A). In B, the percentage of exhaustion/activation markers on CMV-specific T cells is shown (B). Note, CD69 expression is analyzed on CD8 + T cells. The graph in C displays the different phenotypes of CD8 + T cells (Effector: CD45RA + CCR7-, Naïve: CD45RA + CCR7 + , Central memory (CM): CD45RA-CCR7 + , Effector memory (EM): CD45RA-CCR7-) (C). Box plots show the median and quartiles and whiskers display the minimum and maximum (n = 20; note for Tregs, LAG-3, CD69 and graphs in C n = 9). Statistical differences between treatments and untreated cells were determined with Kruskal–Wallis test followed by Dunn’s multiple comparison test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001)