The anti-GFP immune response in mice coinjected with LV-CMV-GFP and GFP-tg APCs. (A) Splenic APCs were isolated from GFP-tg mice, and the FACS profile is shown. (B) C57BL/6 mice were injected with 15 × 106 GFP-tg APCs the day before LV-CMV-GFP injection. Four weeks after injection, the anti-GFP immune response was analyzed in mice injected with vehicle (□), LV-CMV-GFP (▒), or LV-CMV-GFP + GFP-tg APCs (■). CD8+ T cells infiltrating the liver were detected by FACS after mechanical destruction of the tissue. (C) GFP-specific IFN-γ–producing CD8+ T cells present in the spleen were counted by ELISPOT. (D) GFP-specific antibodies present in the sera diluted 10 000-fold were measured by ELISA. Data are expressed as average (± SD). One representative experiment of 2 is presented (3 animals in the control group, which is the same included in Figure 3; and 5 animals in the other groups). (E) Liver sections were analyzed by confocal microscopy after immunostaining for GFP (green) and Topro-3 (blue).
