Figure 7

Characterization of transduction efficiency and immune responses following muscle gene transfer of AAV2-cOva and VSSTSPR-cOva. (A) Transduction efficiency. Gastrocnemius muscles were analyzed for the presence of vector genomes (at day 7; left panel) and corresponding mRNA expression (at days 7, 14 and 21, right panel) using Ova-specific primers. (B) Cellular immune responses. Representative Flow cytometry profiles and bar graphs (n = 5) showing the percentage of Ova-specific CD8⁺ T cells positively stained with H-2K^b/Ova dextramers in spleen (upper panels) and blood (lower panels) 21 days after gene transfer. (C) Humoral responses. Sera were harvested from individual mice at day 21 and analyzed by ELISA to detect anti-Ova IgG. (D) Interferon-γ secreting cells. Bar graph and representative picture showing number of spot forming units (SFU) for 10⁶ splenocytes harvested at day 21 post gene transfer and re-stimulated with the immunodomiant MHC class I restricted Ova_257–264 peptide (left panel) or MHC class II restricted Ova_323–339 peptide (right panel) to stimulate interferon-γ secretion by antigen specific CD8⁺ and CD4⁺ T cells, respectively (n = 5). *p < 0.05; **p < 0.01; ***p < 0.001.