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. 2016 Jan 19;11(1):e0146371. doi: 10.1371/journal.pone.0146371

Fig 3. Recognition by PEPP2-specific CTLs.

Fig 3

Antigen specificity of PEPP2-specific CTLs was analyzed. (A) IFN-γ secretion by CTLs responding to CIR-A24 cells pulsed with PEPP2271-279 or HIV-derived peptide was evaluated by ELISpot assay. The CTLs used in the assays were induced by 3 stimulations and analyzed at the 5th to 7th days after last stimulation.Y-axis indicates the number of spots observed in each well, which contains 3x104 CD8-positive cells. (B) Cytotoxicity of PEPP2-specific CTLs was examined by 51Cr release assay. (C) Cytotoxicity of PEPP2-specific CTLs against CIR-A24 cells pulsed with various concentrations of PEPP2271-279 was examined at E/T ratio of 30. (D) Cytotoxicity of PEPP2-specific CTLs against cancer cell lines KMS11 (HLA-A*24:02-positive, PEPP2-negative), KMS21 (HLA-A*24:02-positive, PEPP2-positive), NB4 (HLA-A*24:02-negative, PEPP2-positive), and K562 (HLA-A*24:02- negative, PEPP2- positive), and PBMCs from HLA-A*24:02-positive HD was assessed by 51Cr release assay. (E) KMS21 cells were pre-treated with anti HLA class I antibody or mouse isotype IgG1 antibody and used for cytotoxicity assay. Experiments were performed in triplicate. *p < 0.05 (Students t-test).