Figure 6.

Binding of the FOXM1‐derived peptides to HLA‐A2. Three FOXM‐1 derived peptides were examined on the actual binding to HLA‐A2 by using a TAP‐deficient, HLA‐A*0201‐positive cell line T2. T2 cells, pre‐incubated overnight at 26°C, were cultured in the presence of 50 μM peptide and 5 μg/ml β₂‐microglobulin in serum‐free medium at 26°C for 1.5 hr and subsequently at 37°C for 18 hr. After the culture, HLA‐A2 expression on the cell surface was measured by flow cytometry with anti‐HLA‐A2 monoclonal antibody BB7.2. A human cytomegalovirus‐derived peptide (QYDPVAALF) and a SARS‐A2‐S‐7 peptide (NLNESLIDL) were used as a negative and a positive control, respectively. The fluorescence index (FI) was calculated from the mean fluorescence intensity (MFI) of HLA‐A2 expressed on T2 cells determined by flow cytometry, using the formula FI = (MFI [T2 cells with FOXM1 peptide]/MFI [T2 cells with CMV peptide]) − 1. BIMAS score predicting HLA‐A2‐binding affinity of the peptides are also indicated.