<p>Splenocytes were isolated on day 17 from mice immunized twice subcutaneously with the peptides 3, 13, 14, 17, 23, or vehicle at day 0 and day 10. The splenocytes were labeled with CFSE and cultured in the presence or absence of 10 µg/mL of the respective peptides for 8 days. Proliferation of CD4<sup>+</sup> (A) or CD8<sup>+</sup> (B) lymphocytes in response to the different CD8 epitopes was analyzed by flow cytometry using anti-CD4 or CD8 antibodies conjugated with PE-Cy5. Results are presented as cell division index (CDI) as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025500#s2" target="_blank">Materials and Methods</a>. (C) The splenocytes were stimulated <i>in vitro</i> with or without the peptides and were stained with anti-CD8 antibody conjugated with FITC, and then fixed and stained for intracellular IFN-γ using PE-conjugated anti-IFN-γ antibody. The percentage of CD8<sup>+</sup> IFN-γ<sup>+</sup> T cells was calculated. *(<i>p</i><0.05) and **(<i>p</i><0.01) indicate they are significantly different from the unstimulated splenocytes. Data is representative of results derived from three independent experiments.</p
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