FIG. 3.

Functional assays for CTL specific for the Env p18-I10 epitope. (A) Seven days after vaccination with 5 × 10⁶ PFU of VSV-Env (n = 2), bulk splenocytes were used in a standard chromium release assay against p18-I10 peptide-pulsed or media-pulsed control P815 target cells. The effector-to-target ratios have been normalized for the percentage of Env-tetramer-positive CD8⁺ cells. One representative assay is shown. Error bars indicate the standard deviation for the assay. (B) Seven days after vaccination with 5 × 10⁶ VSV-Env (n = 2), bulk splenocytes were stimulated with the p18-I10 peptide for 5 h in the presence of brefeldin A and were stained with anti-CD8 antibody and either isotype control, anti-IFN-γ, or anti-TNF-α antibodies and were analyzed by flow cytometry. The gate and corresponding number in the upper right indicate the percentage of cytokine- or isotype control-positive CD8⁺ cells. The results for both mice are shown.