Capsid antigen presentation and killing assay in vitro in AAV-transduced human hepatocytes. HHL5 human hepatocytes were transduced in vitro at increasing MOI with AAV2 or AAV2(Y-F) vectors. (A) Levels of antigen presentation measured with Jurma-VPQ reporter cells 24 hours after vector transduction. Jurma-VPQ cells were added in culture overnight at a ratio of 10:1 reporter:target. RLU, relative light units. (B) CTL assay. HHL5 target cells were transduced overnight and cocultured for 4 hours with effector cells derived by peripheral blood mononuclear cells at an effector:target ratio of 10:1. Percent cytotoxicity was measured relative to a maximum lactate dehydrogenase release (tritonX-treated targets) after background subtraction. All results are reported as average ± standard error of the mean. *P < .05, unpaired, 2-tailed t test. (C) The peptide VPQYGYLTL is shown bound to H2-L^d based on the crystal structure of SPLDSLWWI bound to H2-L^d in Protein Data Bank code 3TJH. VPQYGYLTL is shown as yellow sticks for carbon, blue for nitrogen, and red for oxygen. The peptide VPQYGYLTL is shown bound to HLA-B*-07:02 as modeled from the crystal structure of HLA-B8 from PDB code 3SPV (95.7% identical to HLA-B*07:02:01). The molecular surfaces of H2-L^d and HLA-B*07:02:01 are shown as orange for carbon, blue for nitrogen, and red for oxygen.