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. Author manuscript; available in PMC: 2011 Sep 24.
Published in final edited form as: Vaccine. 2010 Aug 13;28(41):6675–6683. doi: 10.1016/j.vaccine.2010.08.012

FIGURE 4.

FIGURE 4

Long-lived antigen-specific CD8 T cells exhibited the central memory phenotype and were able to secrete multi-cytokines upon peptide reactivation. Naïve C57BL/6 mice were immunized with 669 ng p24 of FKOVA/SVGmu(IN−) (number of mice: n=3) or FKOVA/SVGmu(IN+) (number of mice: n=3) through a footpad injection. Ten weeks later, splenocytes were analyzed by flow cytometry analysis and an ELISPOT assay. (A). Characterization of the surface markers on OVA-specific CD8 T cells with 6-color staining. H-2Kb-SIINFEKL-PE tetramer along with anti-mouse CD8, CD25, CD69, CD44, and CD62L antibodies were used to identify OVA-specific CD8 memory T cells. (B). Quantification of the frequency of OVA-specific CD8 T cells that were displaying the central memory phenotype of CD44+CD62L+CD25−CD69−. (C). Splenocytes were cocultured with the dominant epitope, OVAp1, or the sub-dominant epitope, OVAp3, in the presence of costimulation with an anti-mouse CD28 antibody. Cytokine production was measured by multi-color intracellular staining. (D). Splenocytes were evaluated for their secretion of IL-2 by an ELISPOT assay after restimulation with OVAp1, OVAp2, or OVAp3. The error bars shown in this figure represent deviation between mice within a group.