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 5
The neu-expressing, GM-CSF–secreting vaccine given concurrently with the intact 7.16.4 mAb enhances the effector function and proliferation capability of neu-specific CD8⁺ T cells in vivo. (A) The neu-expressing, GM-CSF–secreting vaccine given concurrently with the intact 7.16.4 mAb increases the number of activated IFN-γ–expressing RNEU_420–429-specific CD8⁺ T cells following treatment. Each neu-N mouse received 2 × 10⁶ Thy1.2 RNEU_420–429-specific CD8⁺ T cells i.v., followed by 1 × 10⁶ 3T3 neu/GM or 3T3/GM cells in each limb s.c. and intact 7.16.4 mAb (100 μg), 7.16.4 F(ab′)₂ (150 μg), or irrelevant IgG (100 μg) i.p. on day 0. Their spleens and VDLNs were harvested on day 4, and CD8⁺ T cells were isolated with the Miltenyi CD8a magnetic beads. The isolated CD8⁺ T cells were then cocultured (1 × 10⁶) with RNEU_420–429-pulsed T2D^q (1 × 10⁶) overnight. Thy1.2 RNEU_420–429-specific CD8⁺ T cells were stained for IFN-γ and analyzed by flow cytometry. The mean fluorescent intensity of IFN-γ in Thy1.2 RNEU_420–429-specific CD8⁺ T cells was also measured. Shown is a representative flow cytometric analysis for 1 mouse per group. This study was performed on a total of 3 mice per group per experiment and was repeated once. The statistical analysis is shown in Table 2. *P < 0.05 as determined by the Mann-Whitney U test compared with 3T3 neu/GM + intact 7.16.4 mAb group. (B) The intact 7.16.4 mAb enhances proliferation of adoptively transferred TCR transgenic T cells in vaccinated neu-N mice. CFSE dilution of Thy1.2 RNEU_420–429-specific CD8⁺ T cells was measured by flow cytometry. Shown is a representative flow cytometric analysis of 1 mouse per group. A total of 3 mice per group were analyzed per experiment, and this experiment was repeated once. The statistical analysis is shown in Table 3.