Figure 3

T-cell-mediated immune response after vaccination with Hsp70-Mage-a1 fusion protein. C57BL/6 mice were immunized with purified GST, Hsp70, Mage-a1, Mage-a1-Hsp70 fusion protein or Mage-a1 + Hsp70 and then administered a booster immunization after one week. Two weeks after the booster, pooled splenocyte cultures were prepared and re-stimulated. A. IFN-γ-producing T cells in splenocytes of immunized mice were quantified by using ELISPOT. Data are presented as mean ± SE of three separate experiments. Statistical analysis by one-way ANOVA revealed that mice vaccinated with Mage-a1-Hsp70 generated more IFN-γ spots than Mage-a1 or Mage-a1 + Hsp70 (* p < 0.001). B. The CTL response induced in splenocytes of immunized mice was determined by using CytoTox 96 nonradioactive cytotoxicity assay. Data are presented as mean ± SE of three separate experiments. Statistical analysis by two-way ANOVA revealed that Mag-a1-specific lysis of CTLs from mice vaccinated with Mage-a1-Hsp70 was higher than those vaccinated with Mage-a1 or Mage-a1 + Hsp70 at all effector: target cell ratios (* p < 0.001).