FIG. 7.

Exposure to MVA-HIV-infected cells inhibits HIV replication in DCs and DC-T-cell cocultures. (A) Impact of MVA-HIV-infected cells on HIV replication in DCs. HeLa cells were infected with MVA-HIV (1 h, MOI = 1) (HeLa-MVA) or mock infected (HeLa), washed extensively, seeded (5 h) to allow MVA-HIV infection, and cocultured with DCs (1 HeLa cell for 10 DCs). After overnight (ON) coincubation, DCs were harvested by gentle pipetting and infected with HIV_NL-AD8 (2 h, high and low doses of 100 and 1 ng of HIV Gag p24/ml/10⁶ cells, respectively), and HIV replication was monitored. (B) Kinetics of HIV replication in DC culture supernatants, assessed by HIV Gag p24 ELISA. Data are means ± SD for duplicates and are representative of 2 independent experiments. (C) Impact of MVA-HIV-infected cells on HIV transfer. HeLa cells or primary myotubes were infected with MVA-HIV or mock infected and were cocultured with DCs as described for panel A. After overnight coincubation, DCs were harvested, infected with HIV_NL-AD8 (2 h, 1 ng of HIV Gag p24/ml/10⁶ cells), washed, and cocultured with activated autologous CD4⁺ T cells (1 DC to 1 T cell) in 96-well plates at 2 × 10⁶ cells/ml. (D) Kinetics of HIV replication in DC-T-cell culture supernatants, assessed by HIV Gag p24 ELISA. As controls, DCs were cultured separately (DC) and CD4⁺ T cells were directly infected with the same HIV dose (T), and HIV replication was monitored. Data are means ± SD for duplicates and are representative of 3 independent experiments.