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Figure 2: HLA-A2 restricted T cell response and cytolytic potential of T cells specific for the EDDR1 derived p15 epitope: PBMC from a healthy HLA-A2+ donor were in vitro stimulated with the synthetic peptide corresponding to the EDDR1 derived p15 epitope. (A) These cells were tested in an ELISpot assay using T2 cells or normal kidney cells (Normal) obtained from an HLA-A2+ healthy donor tissue loaded with an irrelevant synthetic peptide (flu) or p15 peptide. HLA-A2+ cancer cells lines [ovarian cancer cell line SKOV3-A2 and breast cancer cell line MDA-MB231] were pretreated with W6/32 or BB7.2 antibody prior to incubation with p15 specific T cells overnight. Interferon-γ producing cells were quantitated using Immunospot reader. (B) p15 specific T cells were used in an ELISpot assay with the above mentioned targets and the release of Granzyme B was measured as described above. Error bars represent SEM of experimental replicates. |