Fig. 5.

TRAMP-tumor cell vaccination in combination with CTLA-4 blockade leads to T cell responses against both wild-type and mutated SPAS-1 epitopes in vivo. Three C57BL/6 mice received the TRAMPC2-GM/anti-CTLA-4 vaccination regimen described in Methods. After the fifth and last boost with the tumor cell vaccine, their splenocytes were tested for specific IFN-γ production in response to titrated doses of either the mutated (SNC9-H₈) or the wild-type (SNC9-R₈) SPAS-1 peptides in an intracellular cytokine assay (A) and an ELISPOT assay (B). Ten micromolars of the OVA-SL8 peptide was used as irrelevant antigen in the ELISPOT assay. Standard deviations of triplicate cultures are shown. These are representative results of three independent experiments.