Figure 5.

E7-specific CD8⁺ T-cell immunity induced in mice by injection of exosomes from mice inoculated with Nef^mut/E7 DNA vector.

Notes: Donor C57 Bl/6 mice (8 per group) were inoculated two times with the indicated DNA vectors, and 10 days after the last inoculation PBMCs were recovered from retro-orbital bleeding and tested in IFN-γ Elispot assay for the presence of E7-specific CD8⁺ T-cells. In panel A the mean + SD of SFU/10⁵ cells as calculated from a representative of 2 experiments are shown. Two days later, the mice were sacrificed and exosomes were isolated from plasma by differential centrifugation. Equivalent amounts of these exosomes were then inoculated in syngeneic mice (3 per group) three times. Ten days after the last inoculation splenocytes were tested by IFN-γ Elispot assay carried out in triplicate. In panel B the means of SFU/10⁵ cells as calculated from two independent experiments are shown. PMA: cells incubated with 5 ng/mL PMA and 500 ng/mL ionomycin.

Abbreviations: PBMCs, peripheral blood mononuclear cells; PMA, phorbol 12-myristate 13-acetate; SD, standard deviation; SFU, spot-forming unit.