In vitro and in vivo characterization of AAV8-Tetbidir-Alb-IL-12 construction. (a) Analysis of interleukin-12 (IL-12) expression by enzyme-linked immunosorbent assay (ELISA) in the supernatant of hepatic (HepG2, PLC/PRF/5, and HuH-7) and nonhepatic cells (HEK293, HeLa) after pAAV8-Tetbidir-Alb-IL-12 plasmid transfection and administration of doxycycline (Dox). (b, c) For in vivo characterization C57BL/6 female mice received three different doses of AAV8-Tetbidir-Alb-IL-12: 1.5 × 1012, 5 × 1011, 1.5 × 1011 vg/kg (n = 5). Twenty-one days after injection of the rAAV8 vector IL-12 expression was induced by the administration of 50 mg/kg of Dox and then maintained via the administration of Dox in drinking water at a concentration of 2 mg/ml. (b) IL-12 and (c) interferon (IFN)-γ levels in serum were measured by enzyme-linked immunosorbent assay (ELISA) before and 1, 4, 7, and 10 hours after administration of Dox. AAV8, adeno-associated virus serotype 8; Alb, albumin promoter; rAAV, recombinant AAV.