Figure 1.

Establishment of a model of systemic antigen-specific T cell tolerance. (A) Mice were injected i.v. with 10¹⁰ VG of AAV2 or AAV8 expressing luciferase. At day 14 whole body bioluminescence was visualized. (B) Mice were injected i.v. with 10¹¹ VG of AAV-hAAT, and then 2 weeks later challenged i.v. with 10¹⁰ particles of Ad-hAAT. Control mice were injected with PBS. (C) At 9 days after the Ad-hAAT injection, splenocytes or liver leukocytes were stimulated with the hAAT CD8 T cell epitope and subjected to the IFN-γ ELISPOT assay. Background spot-forming unit (SFU) values were subtracted prior to plotting. (D) At 8 weeks after the Ad-hAAT injection, hAAT plasma levels were measured using ELISA. (E) Mice were injected i.v. with 10¹¹ VG of AAV-hAAT or PBS, and then 2 weeks later challenged i.v. with 10¹⁰ particles of Ad-LacZ. Livers were processed for LacZ expression 9 and 28 days later. (F) Mice were injected i.v. with 10¹¹ VG of AAV-hAAT or PBS, and then 2 weeks later challenged i.v. with 10¹⁰ particles of Ad-LacZ. Nine days after the Ad-LacZ injection, splenocytes were stimulated with the LacZ CD8 T cell epitope and subjected to the IFN-γ ELISPOT assay. Background SFU values were subtracted prior to plotting. Data represent groups of three mice in three independent experiments that gave similar results.