Skip to main content
. 2013 Jun 4;8(6):e64365. doi: 10.1371/journal.pone.0064365

Figure 1. T2 binding assays and immunogenicity study.

Figure 1

(A). The HLA-A2 binding affinity of the 5 predicted POTE epitopes was confirmed by T2 binding assay. The calculated FI0.5 for POTE 222, 252, 272, 323 and 553 were approximately 42.4 µM, 1.6 µM, 0.9 µM, 0.8 µM and 5.6 µM, respectively. Binding affinity of the substituted POTE 252 peptides (B), the substituted POTE 553 peptides (C) or the substituted POTE 323 peptides (D) to HLA-A2 molecules. Each assay was performed in triplicate, and data in this figure are representative of two experiments with similar results. Immunogenicity of the POTE 272 epitope in AAD mice (E). Mice were immunized with 50 nmol of POTE 272 in 100 µl of emulsion, and targets were pulsed with 1.0 µM POTE272. Spots were counted by an ELISPOT reader (AID ELISPOT reader system). Figures show numbers of spots per million cells. (F) CTL reactivity on POTE 272 peptide. In a 4 hour 51Cr release assay, CIR.AAD cells were pulsed with 1.0 µM POTE 272 peptide and labeled with 51Cr. After washing three times, target cells were mixed with different numbers of effector cells and then cultured for 4 hours before harvesting.