Figure 6

Targeting cyclin D1 to DCs via CD40 elicits cyclin D1-specific CD4⁺ and CD8⁺T cell responses. (A) 5 x10³ IFN-DCs were loaded with 3 μg/ml recombinant cyclin D1 fusion proteins or control IgG-cyclin D1. After 8 h, CFSE-labeled autologous enriched CD3⁺T cells were co-cultured for 7 days then added the same amount of cyclin D1 fusion protein or control IgG-cyclin D1-loaded IFN-DCs cultured for another 7 days. Cells were boosted by 71 of 15-mer cyclin D1 peptides following 2-day resting in serum-free medium. Intracellular staining of IFN-γ was performed 8 h later. The cells without peptide boosting were used as a negative control. The percentage of IFN-γ⁺CFSE⁻ cells in gated CD4⁺ T cells and IFN-γ⁺Granzyme B⁺ cells in gated CD8⁺ T cells were indicated. This data is a representative of three independent experiments of a healthy donor ND257 (HLA-A*0201). (B) Intracellular staining of IFN-γ/CD107a performed in another healthy donor ND239 (HLA-A*0201) was shown. Percentage of IFN-γ⁺CD107a⁺ cells from CD8⁺ gated is indicated.