FIG. 4.

Inflammatory cytokine production in response to RAHYNIVTF by CD8⁺ T cells is enhanced in the spleens of mice vaccinated with VacLLOE7. Following two i.p. injections 7 days apart with 10⁷ PFU of VacGag, VacE7, VacSigE7LAMP-1, or VacLLOE7, spleen cells from two mice per treatment group were harvested 13 days after the second injection. Whole splenocytes were incubated with 1 μM HPV-16 E7 peptide RAHYNIVTF for 5 h in the presence of a Golgi transport inhibitor, and the levels of intracellular IFN-γ (A) and TNF-α (B) were determined by flow cytometry as described in Materials and Methods. Values shown are percentages of activated CD8⁺ T lymphocytes (gated on CD62L) that are IFN-γ positive. IFN-γ production in the presence of an irrelevant peptide (AMQMLKETI) from HIV Gag was no more than 10% the value shown for each vaccinia virus recombinant expressing E7. Responses shown are representative of three experiments.