Figure 1.

TRBV9*01 is required for optimal recognition by a public TCR that dominates the response to the HPVG viral epitope. (a) Highly conserved V-(D)-J junctional region sequences of TCR α and β chains from CTL clones that recognize HLA-B*3501^HPVG, isolated from three unrelated individuals. The nucleotide sequences are presented, and the one-letter code designating the translated amino acid is shown above the first nucleotide in each codon. Colored areas designate nucleotides of germline origin. (b) HLA-B*3501^HPVG multimer staining, and CD3 and CD8 cell-surface expression for JurkatCD8 cells transduced with the TK3^WT, TK3^Gln55His (two clones), or TK3^Gln55Ala TCR. Cells were stained with OKT3 (anti-CD3), anti-CD8a, or an HLA-B*3501^HPVG multimer at 4°C. Data for the untransfected parental line was left out of the histogram for simplicity. (c) The TCR-transduced JurkatCD8 cells were also stained at 37°C for 4 h with the HLA-B*3501^HPVG pentamer using a range of multimer concentrations (final concentration indicated on the x axis). Mean fluorescence intensity (MFI) of HLA-B*3501^HPVG pentamer staining is shown on the y axis. (d) Activation of the TCR-transduced JurkatCD8 cells with various concentrations of the HLA-B*3501^HPVG pentamer. CD69 up-regulation was used as a marker for cell activation. The binding of HLA-B*3501^HPVG pentamer was performed at 37°C for 4 h. The cells were then washed and costained with APC-labeled anti-CD69 antibody for 30 min on ice. The experiments were conducted at least twice with similar results.