FIG 2.
Myeloid dendritic cells are able to repress immediate early gene expression in coculture with LSEC or MEF. MEF or LSEC were infected with recombinant MCMVs and cocultured with antigen-specific CD8+ T cells or mDC. (A) LSEC infected with MCMVIE2SL-MIEP (MOI of 0.001). Where indicated, LSEC were cocultured with CD4− CD19− CD8+ sorted cells from gBT-I (ratio of 1:1) mice, CD11c+ mDC (ratio of 1:1), or both mDC and CD8 T cells. Virus titer in supernatants on day 5 postinfection was assessed in triplicate, and average values ± SEM are shown. (B) LSEC infected with MCMVr (MOI of 0.01) were cocultured with CD3− Nkp46+ sorted NK cells (ratio of 4:1), CD11c+ mDC (ratio of 4:1), or both NK cells and DC and compared to LSEC monocultures as control. Virus titer in supernatants on day 6 postinfection was assessed in triplicate, and average values ± SEM are shown. (C) LSEC were infected with MCMVr at an MOI of 0.1 and cocultured with increasing proportions of CD11c+ mDC. Each condition was performed in biological triplicates in two independent experiments, and average virus titers in supernatants at day 6 postinfection are shown. Errors bars indicate SEM. (D) MEF were infected with MCMVr at the indicated MOIs and cocultured with CD11c+ mDC at a ratio of 1:4. Supernatants from biological triplicates at the indicated days were titrated on MEF, and graphs show average values ± SEM. (E) Representative bright-field and fluorescence microscopy pictures of MEF infected at an MOI of 0.1 in the presence or absence of CD11c+ mDC. Pictures were taken at day 6 postinfection. DL, detection limit.