Figure 5. EBV and HHV-6A dUTPase DCM induce cytokines that are associated with T_FH-like and T_H1 cell signatures.

(A) Naive CD4⁺ T cells were isolated from PBMCs as described above (Figure 2 and Methods section), cultured in vitro with anti-CD3/anti-CD28 coated beads alone (Beads) or in the presence of DCM (Ctl or EBV dUTPase or HHV-6A dUTPase; 25% vol/vol), and restimulated or not on day 3 with PMA/I. Following treatments, supernatants were examined for IL-21 (A) and IFN-γ (B) concentrations by ELISA. (C and D) ELISA of CXCL13 or IL-10 in supernatants of naive CD4⁺ T cells stimulated for 5 days by plate-bound anti-CD3 and human recombinant ICOS ligand chimera, in the absence (Neg Ctl) or presence of DCM (Ctl, EBV dUTPase, or HHV6A dUTPase) alone or together with IL-12. Treatment of cells with activin A plus IL-12 was used as the positive control, and cells stimulated with beads only were used as the negative control. Data represent the mean ± SEM of n = 6–8 (A, C, and D) and n = 4 (B) independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 of treatments versus negative Ctl or beads by 1-way ANOVA Kruskal-Wallis multiple comparisons test with Dunn’s correction.