Percent CD25⁺ T cells of CD4⁺ DO11.10 TCR⁺ cells in DO11.10-tg RAG-2^−/− BALB/c mice as determined by flow cytometry. (A) Scatter graphs show staining of CD25 and DO11.10 TCR for gated CD4⁺ cells. This example represents inguinal lymph node cells from control mice and animals that received hepatic gene transfer with 1 × 10¹² vg AAV-EF1α-ova vector. Mice were killed 8 weeks after gene transfer. Antibody stainings were FITC-conjugated KJ1-26 (for DO11.10 TCR) and PE-conjugated CD25. (B) Summary of results for different lymphoid organs including fold difference between groups. Mice had received no gene transfer or AAV-EF1α-GFP or AAV-EF1α-ova. Results from spleens and thymus were average (± SD), while lymph nodes cells were pooled prior to flow cytometry (n = 5/cohort). *P < .01 compared with control groups.