Figure 7

Dendritic cells generated in roller bottles stimulate autologous peptide-specific T-cell responses. Dendritic cells from HLA-A*0201⁺ normal donor leukopheresis products were differentiated and matured with ITIP in roller bottles or static flasks as before. The floating cells were harvested, pooled, irradiated (20 Gy), and pulsed with HLA-A*0201 epitopes from flu, EBV, and CMV (see Materials and Methods for details). The DC were washed and added to 500,000 T-cell-enriched autologous PBMC in anti-IFN-γ antibody-coated ELISPOT plates in the numbers indicated. Each assay was run in triplicate. The plates were harvested after overnight culture and developed. Shown is the image taken of the developed ELISPOT plate (A) and corresponding graphical representation of the number of spots per 500,000 input responder cells under the different conditions (B). Dendritic cells without peptide pre-pulsing were used as controls. The results are representative of two similar experiments.