Figure 8.

Expression of immune checkpoint factors on the CD4 and CD8 T cells activated by CD3 and CD28 ligation and supernatants from the natural killer (NK)–dendritic cell (DC) cross talk assay. DCs and NK cells from the same donor were exposed to 1 μg/ml F-HIV, complement-opsonized HIV (C-HIV), CI-HIV, PHA, or mock treated for 24 h. The experiment was replicated four times using cells derived from four different donors. Allogeneic T cells were stimulated by CD3/CD28 ligation in the presence of supernatants from these cultures. (A,D) The CD4 T cell (A) and CD8 T cell (D) phenotype generated was assessed using flow cytometry. Heat maps of the percentage of CD4 cells or CD8 T cells positive for phenotypic markers was created. The number of CD4 T cells (B) or CD8 T cells (E) positive for CD38, and the number of cells coexpressing PD-1, TIM-3, and LAG-3 were evaluated. (C,F) Gating strategy for flow cytometry analysis of a representative sample. Boxplots show mean with Tukey error bars. One-way ANOVA followed by a Bonferroni posttest was used to test for statistical significance (*p < 0.05, N = 4).