FIG 2.

Gating strategy used to detect CD8 and CD4 T cells producing multiple cytokines in the splenocyte populations of vaccinated mice. Mice were vaccinated with three 10-μg doses or three 25-μg doses of DNA intradermally, and splenocytes were harvested on day 14 after the final vaccination for ICS. Cytokine profiles were determined by using flow cytometry. Splenocytes were gated on the lymphocyte population, followed by doublet discrimination, and then gated on CD3⁺ CD44⁺ cells and finally CD4⁺ or CD8⁺ cells to assess the frequency of IFN-γ, TNF-α, and IL-2. Representative blots for IFN-γ-, TNF-α-, and IL-2-positive cells are shown.