Dendritic cells pulsed with hsp70-peptide complexes derived from human hepatocellular carcinoma induce specific anti-tumor immune responses

Figure 4 Secretion of IFN-γ by T cells induced with autologous DCs pulsed with hsp70-peptide complexes derived from SMMC-7721 cells. 1 × 10⁵ DCs were incubated with 20 μg/mL hsp70-peptide complexes derived from SMMC-7721 for 4 h at 37°C and then cocultured with 1 × 10⁶ autologous T cells in each well of 24-well culture plate for 7–10 d in the presence of 20 U/mL human IL-2. The stimulated T cells were harvested by density gradient centrifugation and their IFN-γ production was determined by ELISPOT assay (indicated as hsp70 stimulation). As a control, T cells cocultured with unpulsed DCs (indicated as DC control) or by themselves (indicated as T control) were also used as effectors. The results are expressed as mean±SD and each value represents the mean of three replicates.