Figure 2.

AdV_OVA stimulates ovalbumin (OVA)-specific CD8⁺ cytotoxic T lymphocyte (CTL) responses. (a) The AdVova-immunized C57BL/6 mouse tail blood samples harvested on different days after the immunization of OVA-Texo were stained with PE-H-2K^b/OVAI peptide tetramer and fluorescein isothiocyanate (FITC)-anti-CD8 antibody (Ab), and then analyzed by flow cytometry. (b) The tail blood samples of AdVova-immunized C57BL/6 mice were harvested on day 11 after the immunization and stained with PE-H-2K^b/OVAI peptide tetramer and FITC-anti-CD8 Ab, and then analyzed by flow cytometry. The value in each panel represents the percentage of OVA-specific (tetramer-positive) CD8⁺ T cells vs the total CD8⁺ T-cell population. The value in parenthesis represents s.d. (c) In vivo cytotoxicity assay. Six days after the immunization, the immunized mice were i.v. injected with 2 × 10⁶ cells containing a 1:1 mixture of CFSE^high- and CFSE^low-labeled splenocytes that had been pulsed with OVAI or Mut1 peptides, respectively. After 16 h, the spleens of immunized mice were removed and the percentages of the residual CFSE^high (H) and CFSE^low (L) target cells remaining in the recipients’ spleens were analyzed by flow cytometry. The value in each panel represents the percentage of CFSE^high vs CFSE^low target cells remaining in the spleen. The value in parenthesis represents the s.d. ^*P<0.05 vs cohorts of the control AdVnull group (Student's t-test). One representative experiment of three is shown. AdV, adenovirus; CFSE, carboxyl-fluorescein succinimidyl ester.