Antibody association with HER-2/neu–targeted vaccine enhances CD8⁺ T cell responses in mice through Fc-mediated activation of DCs
J. Clin. Invest. Peter S. Kim, et al. 118:1700 doi:10.1172/JCI34333 [Go to this article.]

Figure 4
CD11c⁺ DCs isolated from mice treated with the neu-expressing, GM-CSF–secreting vaccine given concurrently with intact 7.16.4 mAb induce efficient proliferation of naive neu-specific CD8⁺ T cells. (A) Naive RNEU_420–429-specific CD8⁺ T cells proliferated most vigorously in response to CD11c⁺ DCs isolated from mice treated with the neu-expressing, GM-CSF–secreting vaccine + intact 7.16.4 mAb. Administration of vaccine cells and antibodies was similar to that described in Figure 3A. On day 4 after vaccination, the spleen and VDLNs were harvested and DCs were isolated using the Miltenyi CD11c magnetic beads. Naive RNEU_420–429-specific CD8⁺ T cells were isolated, labeled with CFSE, and then cocultured with CD11c⁺-isolated DCs for 3 days. CFSE dilution was measured by flow cytometry. n = 5 mice per group. This analysis was repeated at least twice. (B) Both CD8⁺ and CD8^– DCs proliferated RNEU_420–429-specific CD8⁺ T cells most efficiently when the intact 7.16.4 mAb was administered with the neu-targeted vaccine. Administration of vaccine cells and antibodies was similar to that described in Figure 3A. On day 4, splenic CD11c⁺ DCs were isolated from each group, followed by FACS of CD8⁺ and CD8^– DCs. Subsequently, the DEAD assay was performed as described in A. n = 10 mice per group. This analysis was repeated at least twice. *P < 0.05 versus intact 7.16.4 mAb + neu-targeted vaccine, Mann-Whitney U test.